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Chromatography Applications
HPLC | IC | SPE
discoverysciences.com
Applications
HPLC Applications . . . . . . . . . . . . . .
Pharmaceuticals 1–17
Nutraceuticals/Vitamins 18–24
Food & Beverage 25–38
Life Science 39–52
Environmental 53–57
Miscellaneous 58–63
IC Applications . . . . . . . . . . . . . . . .
Anions 64–66
Cations 67–68
SPE Applications . . . . . . . . . . . . . . . 69–79
Pharmaceuticals 69
Nutraceuticals 70–71
Food & Beverage
Life Science 74
Environmental 75–79
72–73
64–68
1–63
0 2 4 6 8
1
2
3 4
0 2 4 6 8 10 12 14 M0 2
1
2 3
4
5
6
4 6 8
0
2
1
2.5 5 7.5 10 12.5 15 17.5
Column: Alltech® Platinum™ C18, 5µm, 150 x 4.6mm
(Part No. 32043)
Mobile Phase: 0.02M KH2PO4:Methanol:Acetonitrile:
pH4.5 (40:50:10)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Naproxen 2. Acemetacin 3. Indomethacin 4. p-Xylene
Column: Alltech® Prevail™ Amide, 5µm, 150 x 4.6mm
(Part No. 88660)
Mobile Phase: 0.02M K2HPO4, pH7.0:Acetonitrile (75:25)
Flow Rate: 1.0mL/min
Detector: UV at 210nm
1. Aspirin 2. Acetaminophen 3. Naproxen 4. Fenoprofen
938510124
1. Aspirin 2. Acetaminophen 3. Naproxen 4. Fenoprofen 5. Ibuprofen 6. Diclofenac
Column: ® Alltima C8, 5µm, 150 x 4.6mm (Part No. 88072)
Mobile Phase: A: 0.05M K2HPO4, pH3.3 B: Methanol
C: Acetonitrile
Gradient: Time: 0 2 5
%B: 10 10 30
%C: 10 10 25
Flow Rate: 2.0mL/min
Detector: UV at 280nm
Column: HP C8, 5µm, 150 x 4.6mm
(Part No. 87751)
Mobile Phase: 0.05M Phosphate Buffer pH3.0:Methanol (80:20)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Procainamide 2. Acetaminophen 3. Theophylline 4. Caffeine
Column: GraceSmart™ C18, 5µm, 150 x 4.6mm
(Part No. 5138812)
Mobile Phase: Phosphoric Acid in Water, pH2.5:Acetonitrile (65:35)
Flow Rate: 2mL/min
Detector: UV at 214nm
Injection Vol: 5µL
1. Valerophenone 2. Ibuprofen
Analgesics
Anti-Inflammatories
AnalgesicsAnalgesics
Basic Drugs
Ibuprofen (U.S.P.)
8156 10702
10374
0 1
1
2 3 Min.
1
1
1
1
30°C
32.5°C
35°C
37.5°C
40°C
42.5°C1
1. Ibuprofen (100ppm)
Column: µm, 2.1 x 100mm
(Part No. 87669)
Mobile Phase: Acetonitrile: 0.1%TFA in Water (80:20)
Flow Rate: 0.3mL/min
Column Temp: Ambient
Semi-Volatiles
Detector: ELSD
hplc applicationspharm
aceuticals
1 www.discoverysciences.com
®
®
Alltech
® Alltima ®Alltech
HP C18, 3HP C18, 3® Alltima ®Alltech
10 Min. 20 Min.
10 Min. Min.
Min.
0 1 2 3 4 5 Min.
1
2
3
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: Water:Acetonitrile (10:90)
Flow Rate: 1.0mL/min
Detector: UV at 254
1. Methyl Paraben 2. Propyl Paraben 3. Hydrocortisone Acetate
9463
Hydrocortisone Acetate Cream
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Phosphate Buffer:Methanol (80:20)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
10770
Paracetamol Effervescent Tablets on Chinese Pharmacopeia Assay
Analgesics
mA
U
2000
1250
1000
1500
1750
750
500
250
0
5 6 7 83 41 2 9 10 Min.0
1. Paracetamol
0 1 2 3 4 5 6 7 8 9 10 Min.
2
1
Alltima HP
HILIC Columns
1
2
Reverse-Phase
C18 Columns
1. Albuterol
2. Bamethan
Basic Drugs
Column: Alltech® Prevail™ C18, 3µm, 53 x 7mm Rocket™
(Part No. 99279)
Mobile Phase: Water:Methanol (10:90)
Flow Rate: 4.5mL/min
Detector: UV at 254
1. Hydrocortisone 2. Testosterone 3. Progesterone
9392
Steroids
Steroids
10391
Column: HP CN, 5µm, 150 x 4.6mm
(Part No. 87769)
Mobile Phase: Hexane:Isopropanol (93.7:6.3)
Flow Rate: 2.0mL/min
Detector: UV at 254nm
1. Deoxycorticosterone Acetate
2. Hydrocortisone Acetate
3. Cortisone Acetate
4. Cortisone
5. Prednisone
6. Hydrocortisone
7. Prednisolone
Corticosteroids
Column: Prevail™ C18 Column, 5µm, 4.6 x 150mm
(Part No. 99208)
Alltima HP HILIC Column, 5µm, 4.6 x 150mm,
(Part No. 86464)
Mobile Phase: 0.05M Ammonium Formate, pH 3.0:CH3CN (88:12)
Acetonitrile:0.05M Ammonium Formate, pH 3.0 (85:15)
Flow Rate: 1.0mL/min
Detector: UV at 220nm
10949
hplc applicationspharm
aceutic
als
2www.discoverysciences.com
® Alltima ®
Alltech
®
®
1
2
0 20 Min.102 4 6 8 12 14 16 18
0 2
1
2
3
4
5
6
7
4 6 Min.
0 1 2 3 4 5 6 7 8 9 Min.
1
2
3
8957
Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™
(Part No. 50605)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.1% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 6
%B: 10 90
Flow Rate: 2.5mL/min
Detector: ELSD
1. Theophylline 2. Acetylsalicylic Acid 3. Propranolol 4. Nortriptyline 5. Trimipramine
Column: Silica, 5µm, 300 x 4.6mm (Custom)
Mobile Phase: Acetonitrile:Chloroform:i-Butyl Alcohol:Water:Glacial
Acetic Acid (85:8:5:2:0.05)
Flow Rate: 2.0mL/min
Detector: UV at 254nm
1. Triazolam 2. Alprazolam
8691
Column: Alltech® Platinum™ EPS C18, 3µm, 53 x 7mm Rocket™
(Part No. 50573)
Mobile Phase: 0.05M Ammonium Acetate, pH5.5:Acetonitrile (65:35)
Flow Rate: 3.0mL/min
Detector: UV at 254nm
9105
10373
Column: CN, 3µm, 150 x 4.6mm
(Part No. 81189)
Mobile Phase: 0.01M KH2PO4, pH2.6:Acetonitrile (50:50)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Desipramine 2. Nortryptyline 3. Diphenhydramine 4. Amitriptyline
7368
1. Diphenhydramine 2. Nortriptyline 3. Amitriptyline
Column: Alltech® Alltima HP C18 Amide, 5µm, 150 x 4.6mm
(Part No. 87733)
Mobile Phase: 0.025M Potassium Phosphate Buffer, pH7.0:Methanol
(15:85)
Flow Rate: 1mL/min
Detector: UV at 254nm
Column: Alltech® Alltima HP C8, 5µm, 150 x 4.6mm
(Part No. 87751)
Mobile Phase: A: 0.025M Potassium Phosphate Buffer, pH7.0
B: Methanol
Gradient: Time: 0 8 20
%B: 60 75 75
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Fluphenazine HCl 2. Chlorpromazine HCl 3. Phenothiazine HCl
1. Chlordiazepoxide Degradant 2. Nitrazepam 3. Norchlordiazepoxide 4. Nordiazepam
Universal High Speed Gradient
10353
Benzodiazepines and Metabolites
Tricyclic Antidepressants
Alprazolam and Triazolam by the USP XXII Method
Antidepressants
Basic Drugs
Tricyclic Amines
5. Chlordiazepoxide 6. Chlordiazepoxide Degradant 7. Diazepam
hplc applicationspharm
aceuticals
3 www.discoverysciences.com
® Alltima ®Alltech ® Alltima ®Alltech
Alltima® ®
®
Min. Min.
Min. Min.
Min. Min.
3
12
3
4
0 1 2 3 4 5 6 7 8 Min.
1
2
3
4
0 1 2 3 4 5 6 7 8 Min.
2
3
1
45
6 7
0 2 4 6 8 10 12 14 16 18 Min.
1072010719
1. Uracil 2. Propanolol 3. Naphthalene 4. Dipropylphthalate
Column: Grace® VisionHT C18, 1.5µm, 50 x 2.0mm
(Part No. 5139605)
Mobile Phase: 20mM Potassium Phosphate, pH2.5:Methanol (40:60)
Flow Rate: 0.5mL/min
Detector: UV at 280nm
Temperature: Ambient
PropanololPropanolol
1. Uracil 2. Propanolol 3. Naphthalene 4. Dipropylphthalate
Column: Grace® VisionHT C18-P, 1.5µm, 50 x 2.0mm
(Part No. 5139606)
Mobile Phase: 20mM Potassium Phosphate, pH2.5:Methanol (40:60)
Flow Rate: 0.5mL/min
Detector: UV at 280nm
Temperature: Ambient
Column: Jones Genesis® C18, 4µm, 150 x 4.6mm
(Part No. FM15960E)
Mobile Phase: Acetonitrile:50mM KH2PO4, pH2.4 (35:65)
Flow Rate: 2.0mL/min
Detector: UV at 254nm
Basic Drugs
1. Amiloride 2. Propranolol 3. Chloramphenicol 4. Carbamazepine 5. Trimipramine 6. Trifluoperazine
10604
Column: Alltech® Alltima HiLoad, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: Water:Acetonitrile (70:30)
Flow Rate: 1.0mL/min
Detector: ELSD
1. Phenobarbital 2. Butabarbital 3. Butalbital 4. Pentobarbital 5. Amobarbital 6. Mephobarbital 7. Secobarbital
Barbiturate Mix
Antidepressants
BarbituratesBarbiturates
Column: C18, , 50 x 2.1mm
Mobile Phase: 40% CH 3CN:50mM Ammonium Acetate, pH5.4
Flowrate: 0.5mL/min
Detector: UV at 230nm
Phenobarbital
Amobarbital
Mephobarbital
5
1. Barbital
2.
3. Talbital
4.
5.
0 1 2 Min.
1
2
3
4
0 1 2 3 Min.
1
23
1. Acetylsalicylic Acid 2. Diphenhydramine 3. Trimipramine
Column: Alltech® Alltima µm,
7 x 33mm Rocket™ (Part No. 87671)
Mobile Phase: A: 0.1% TFA in Water B: 0.1% TFA in Acetonitrile
Gradient: Time: 0 3
%B: 10 90
Flow Rate: 2.5mL/min
Column Temp: 30°C
Stable Baselines with Fast, Steep Gradients
10658
9433
3µmAlltech® Alltima
hplc applicationspharm
aceutic
als
4www.discoverysciences.com
® ®
HP C18, 3®
® ®
pharm
aceuticals
Barbiturates
Column: Platinum™ EPS 100Å C18, 5µm, 150 x 4.6mmMobile Phase: 0.010M Sodium Acetate, pH4.0 Acetonitrile
(75:25)Flowrate: 1.0mL/minDetector: UV at 230nm
Column: Platinum™ EPS 100Å C18, 3µm, 53 x 7mmMobile Phase: 0.010M Sodium Acetate, pH4.0 Acetonitrile
(75:25)Flowrate: 3.0mL/minDetector: UV at 230nm
1. Barbital2. Butalbital3. Aprobarbital4. Talbutal5. Butabarbital6. Phenobarbital7. Amobarbital8. Mephobarbital
Run TimeReducedby 70%
7
12
3
456
8
0 4
12
3
4 5 67
8
8 12 Min.
ock
o o
0 1
12
3
4 5 67
8
2 3 4 Min.
8684
Rocket™ Column
0 2 4 6 8 10 12 14 16 18 20 Min.
2
1
UV at 230nm
21
ELSD
1. Mephobarbital (500ppm) 2. Secobarbital (500ppm)
10616
Column: Alltech® Alltima µm, 4.6 x 250mm
(Part No. 87698)
Mobile Phase: Acetonitrile:Water (30:70)
Flow Rate: 1.0mL/min
Column Temp: 45°C
1:1 Mixture
10617
10375
Column: Alltech® Alltima HP C8, 5µm, 250 x 4.6mm
(Part No. 87751)
Mobile Phase: 0.05M Ammonium Acetate:Acetonitrile (78:22)
Flow Rate: 1.0mL/min
Detector: UV at 230nm
1. Barbital 2. Phenobarbital 3. Talbutal 4. Amobarbital 5. Mephobarbital
Barbiturate Mix
Standard Platinum™ C18
Platinum™ EPS C18
Vasodilators
Column: 5µm, 4.6 x 150mm
Mobile Phase: 0.025M KH2PO4, pH 3.0:Methanol (50:50)
Flow Rate: 1.0mL/min
Detector: UV at 220nm
1. Pindolol 2. Pentoxifylline 3. Isoxsuprine 4. Nifedipine Degradant 5. Nifedipine
8387
8388
Vasodilators
8685
Barbiturates
hplc applications
5 www.discoverysciences.com
HP C18 HL, 5®®
4
pharm
aceutic
als
hplc applications
10348
10349
Column: ® Alltima HP EPS C18, 5µm, 150 x 4.6mm
(Part No. 87715)
Mobile Phase: 0.025M Ammonium Acetate:Acetonitrile (70:30)
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 HiLoad, 5µm, 150 x 4.6mm
(Part No. 87697)
Mobile Phase: 0.025M Ammonium Acetate:Acetonitrile (70:30)
Flow Rate: 1.0mL/min
Detector: ELSD
Carbamazepine
1. Ethotoin 2. Phenytoin
Mephenytoin Carbamazepine
Column: Alltech® Prevail™ Phenyl, 5µm, 150 x 4.6mm
(Part No. 99239)
Mobile Phase: A: Water B: Acetonitrile
Gradient: Time: 0 15
%B: 25 60
Flow Rate: 1.5mL/min
Detector: UV at 254
Anticonvulsants
Anticonvulsants
Anticonvulsants
Anticonvulsants
9394
1. Ethotoin 2. Mephenytoin 3. Carbamazepine 4. Diazepam
0 2 4 6 8 10 12 14 16 18 20 Min.
1
2
3
4
1. Ethotoin 2. Phenytoin 3. Mephenytoin 4.
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.
1
2
3 3.
4.
1. Ethotoin
2. Phenobarbital
3. Methsuximide
4. Mephenytoin
5. Carbamazepine
6. Diazepam
123
4
5
6
0 20 Min.4 8 1612
Column: Phenyl,
Mobile Phase:
Gradient 0 20 25 30
%B: 15 60 60 15
Flow Rate: 1.5mL/min
Detector: UV at 254nm
5,5-DiphenylhydantoinCarbamazepine
Column: Platinum™ EPS 100Å C18, 3µm,53 x 7mm Rocket™ Column
Mobile Phase: 0.025M Ammonium Acetate, pH 5.5: Acetonitrile (70:30)Flowrate: 3.0mL/minDetector: Alltech ELSD
1. Ethotoin2. Mephenytoin3.
4.
0 1
1
2
2
3
4
3 Min.
7233
8946
A: Water B: Acetonitrile: Time:
Anticonvulsants
Anticonvulsants
6www.discoverysciences.com
®
®® Alltima HP EPS C18, 5µm, 150 x 4.6mm®Alltech ® Alltima
®® Alltima ®Alltech 250 x 4.6mm5µm,
pharm
aceuticals
hplc applications
Column: Alltech® Alltima C8, 5µm, 250 x 4.6mm (Part No. 88076)
Mobile Phase: A: 0.05M Potassium Phosphate, pH3.2
B: Acetonitrile
Gradient: Time: 0 5 20 25
%B: 25 25 70 70
Flow Rate: 1.3mL/minDetector: UV at 220nm
1. Tripelennamine 2. Triprolidine 3. Cyclizine 4. Chlorcyclizine 5. Meclizine
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: Acetonitrile:0.1% Ammonium Carbonate, pH6.5 (80:20)
Flow Rate: 1.0mL/min
Detector: ELSD
102757232
Column: Vydac® Denali®, 5µm, 150 x 4.6mm (Part No. 238DE5415)
Mobile Phase: A: Acetonitrile:Water (20:80)
B: Acetonitrile:Water (95:5)
Gradient: Time: 0 8
%B: 40 90
Flow Rate: 1.0mL/min
Detector: UV at 280nm
Antihistamines
Antihistamines
OTC Antihistamine and Excipients
Estrogens
Estrogens
0 1 2 3 4 5 6 7 8 9 10 11 Min.
1
3
4 5
1. Diphenhydramine 2. PEG, Gelatin 3. Sorbitan 4. Glycerol 5. Sorbitol
1. Estriol 2. 17 -Ethylestradiol 3. Estrone 4. Diethylstilbestrol 5. Hexestrol 6. Product of Diethylstilbestrol Decomposition
7 www.discoverysciences.com
®
0
12
3
4
5
2 4 6 8 10 12 Min.
1 + 2
3
0 0.5 1 1.5 2 2.5 3 Min.
1 2
3
0 0.5 1 1.5 2 2.5 3 Min.
pharm
aceutic
als
hplc applications
10118
1. p-Aminobenzoic Acid 2. Benzocaine 3. Tetracaine 4. Dibucaine 5. Butacaine
Column: Alltech® Alltima C8, 5µm, 150 x 4.6mm (Part No. 88072)
Mobile Phase: A: Water B: Methanol C: Acetonitrile
Gradient: Time: 0 7
%B: 15 45
%C: 15 35
Flow Rate: 1.5mL/min
Detector: UV at 230nm
Column: Grace® VisionHT C18, 1.5µm, 50 x 2.0mm
(Part No. 5139605)
Mobile Phase: 25mM Potassium Phosphate, pH7.0:Methanol (30:70)
Flow Rate: 0.5mL/min
Detector: UV at 230nm
1. Procaine 2. Lidocaine 3. Tetracaine
10737 10738
Column: Grace® VisionHT C18-P, 1.5µm, 50 x 2.0mm
(Part No. 5139606)
Mobile Phase: 25mM Potassium Phosphate, pH7.0:Methanol (30:70)
Flow Rate: 0.5mL/min
Detector: UV at 230nm
1. Procaine 2. Lidocaine 3. Tetracaine
Anesthetics
Anesthetic Mix
Basic Pharmaceuticals Basic Pharmaceuticals
1. Procaine 2. Benzocaine 3. Mepivacaine 4. Lidocaine 5. Tetracaine 6. Dibucaine
Column: Vydac® Denali®, 5µm, 150 x 4.6mm (Part No. 238DE5415)
Mobile Phase: Methanol:50mM K2HPO4 in Water, pH7.0 (60:40)
Flow Rate: 1.0mL/min
Detector: UV at 220nm
Anesthetics
1. Hydromorphone 2. Oxycodone 3. Codeine 4. Ethylmorphine 5. Nalorphine
Column: Vydac® Denali®, 5µm, 250 x 4.6mm (Part No. 238DE54)
Methanol:20mM K2HPO4 in Water, pH7.0 (50:50)
Flow Rate: 1.0mL/min
Detector: UV at 254
Opiates
Opiates
8www.discoverysciences.com
® ®
®
0 1
1
2
2
3
3
4
5
6
4
5 Min.
0 2 4 6 8 10 12 14 16 18 20 22 24 Min.
1
2
3
4
0 2 4 6 Min.
1
2
3
5
4
pharm
aceuticals
hplc applications
Column: Alltech® Prevail™ C18, 3µm, 53 x 7mm (Part No. 99279)
Mobile Phase: 0.025M KH2PO4, pH3.0:Acetonitrile (70:30)
Flow Rate: 4.0mL/min
Detector: UV at 220
1. Metoprolol 2. Oxprenolol 3. Propranolol
Column: ZirChrom® PBD, 3µm, 150 x 4.6mm (Part No. ZR03-1546)
Mobile Phase: 0.01M Triethylamine, 0.05M Tetramethylammonium
Hydroxide, pH12.2:Acetonitrile (65:35)
Flow Rate: 1.0mL/min
Detector: UV at 254
10354
10161
Column: Alltech® Alltima HP CN, 5µm, 150 x 4.6mm
(Part No. 87769)
Mobile Phase 0.02M KH2PO4, pH3.0:Acetonitrile (70:30)
Flow Rate: 1.0mL/min
Detector: UV at 218nm
9395
10356
1. Labetalol 2. Acebutolol 3. Oxprenolol 4. Lidocaine 5. Alprenolol 6. Propranolol
8160
Column: Alltech® Alltima HP C18 Amide, 5µm, 150 x 4.6mm
(Part No. 87733)
Mobile Phase: 0.025M Potassium Phosphate, pH2.8:Methanol (90:10)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Codeine 2. Strychnine 3. Quinine 4. Quinidine
1. Diltiazem 2. Verapamil 3. Nicardipine 4. Nifedipine
Column: Alltech® Platinum™ C18, 5µm, 150 x 4.6mm
(Part No. 32043)
Mobile Phase: 0.05M Ammonium Dihydrogen Orthophosphate, pH4.5:
Methanol:Acetonitrile (40:50:10)
Flow Rate: 1.0mL/min
Detector: UV at 230nm
1. Barbital 2. Phenobarbital 3. Butalbital 4. Oxazepam 5. Triazolam
SedativesSedatives/Hypnotics
Basic Drugs
Anti-Hypertensives
Antihypertensives
Calcium Channel Blockers Drug Mix
ELSD
UV
0
1
1 2 3 4 5 6 7 Min.
1. Ethinamate (5.0 mg/mL)
Column: Alltech® Alltima µm, 10 x 150mm
(Part No. 81102)
Mobile Phase: Water:Acetonitrile (55:45)
Flow Rate: 5.0mL/min
Column Temp: 25°C
Ethinamate
10438
9 www.discoverysciences.com
C18, 5
® ®
®
pharm
aceutic
als
hplc applications
Column: Alltech® Platinum™ EPS C18, 100Å, 5µm, 150 x 4.6mm (Part No. 32214)Mobile Phase: Water:Acetonitrile:Methanol:Glacial Acetic Acid (60:10:30:1)Flow Rate: 1.0mL/minDetector: UV at 254nm
Methylxanthines
Amphetamines
Anti-Asthmatics
0 1
1
2
2
3
3 4
4 Min.
1001410427
Column: Alltech® Alltima Silica, 3µm, 100 x 4.6mm (Part No. 81404)Mobile Phase: 1%Ammonium Hydroxide in Methanol:Acetonitrile (25:75)Flow Rate: 2.0mL/minDetector: UV at 280nm
1. 2,3-MDA 2. 3,4-MDA 3. 2,3-MDMA 4. 3,4-MDMA
Column: Alltech® Alltima HP Silica, 3µm, 150 x 4.6mm (Part No. 87786)Mobile Phase: 1%Ammonium Hydroxide in Methanol:Acetonitrile (75:25)Flow Rate: 1.0mL/minColumn Temp: 25°CDetector: UV at 280nm
1. 2,3-MDA 2. 3,4-MDA 3. 2,3-MDMA 4. 3,4-MDMA
MDA & MDMA AnalogsMDA and MDMA Analogs
8384
1. Theobromine 2. Theophylline 3. 8-Chlorotheophylline 4. Caffeine 5. 7-(2-Chloroethyl)theophylline
0 1
1
2
3 4 5
2 Min.
1. Xanthine 2. Theobromine 3. Theophylline 4. Caffeine 5. 8-Chlorotheophylline
Column: Platinum™ C18-EPS 100Å, 3µm, 7 x 33mm Rocket™ Mobile Phase: 0.010M Sodium Acetate, pH 4.0:Methanol (70:30) Flow Rate: 5.0mL/min Detector: UV at 254nm
Rocket™ Columns for High Throughput
with Conventional LC
8679
10www.discoverysciences.com
® ®
mA
U
140
160
180
120
80
100
40
60
0
20
2 4 6 8 10 12 14 16 Min.0
10708
Nifedipine
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: 0.39% Sodium Phosphate Monobasic, pH4.5:
Acetonitrile (35:65)
Flow Rate: 1.5mL/min
Detector: UV
Statins
Simvastatin Tablets (U.S.P.)
at
ph
arm
ace
uti
ca
lshplc applications
10246
10262
10265
1. Amiloride HCl Hydrate 2. Acetazolamide 3. Hydrochlorothiazide 4. Triamterene 5. Chlorthalidone 6. Furosemide 7. Hydroflumethiazide 8. Clopamide 9. Ethacrynic Acid 10. Trichloromethiazide 11. Probenecid 12. Indapamide 13. Bumetanide 14. Bendroflumethiazide 15. Spironolactone
1. Amiloride HCl Hydrate 2. Acetazolamide 3. Hydrochlorothiazide 4. Chlorthalidone 5. Hydroflumethiazide 6. Clopamide 7. Trichloromethiazide 8. Indapamide 9. Bendroflumethiazide 10. Bumetanide 11. Probenecid 12. Spironolactone
1. Acetazolamide 2. Hydrochlorothiazide 3. Triamterene 4. Hydroflumethiazide 5. Clopamide 6. Trichloromethiazide 7. Indapamide 8. Bumetanide 9. Spironolactone
Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™ (Part No. 50605)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in 0.025M Ammonium Acetate B: 0.1% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 4
%B: 20 95
Flow Rate: 3.5mL/min
Detector: UV at 254nm
Column: Alltech® Alltima C18, 5µm, 250 x 4.6mm (Part No. 88056)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in 0.025M Ammonium Acetate B: 0.1% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 10 15
%B: 25 95 95
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Alltima C18, 5µm, 250 x 4.6mm (Part No. 88056)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in 0.025M Ammonium Acetate B: 0.1% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 30
%B: 25 65
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Diuretics Diuretics and Uricosuric Agents
Doping Agents
Diuretics — High Throughput
11 www.discoverysciences.com
®
®
®
0 2
1
2
3 4
5
4 6 8 10 Min.
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.
1
2 34
0 1
1
2
2
3
3
4
4
5 6 7 8 9 Min.
0 1 2 3 4 5 Min.
1
2
pharm
aceutic
als
hplc applications
Column: Alltech® Platinum™ EPS C18, 5µm, 150 x 4.6mm
(Part No. 32043)
Mobile Phase: 0.025M KH2PO4, pH3.0:Acetonitrile (90:10)
Flow Rate: 1.0mL/min
Detector: UV at 230nm
1. Sulfanilamide 2. Cefaclor 3. Cefatrizine 4. Thiamphenicol 5. Cefotaxime
1. Oxytetracycline Dihydrate 2. Tetracycline HCl 3. Meclocycline Sulfosalicylate Salt
Column: Alltech® Alltima C18, 3µm, 100 x 4.6mm (Part No. 81382)
Mobile Phase: A: 0.05M KH2PO4, pH3.0 B: Acetonitrile
Gradient: Time: 0 10
%B: 15 40
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Column: Alltech® Alltima C18, 5µm, 250 x 4.6mm (Part No. 88056)
Mobile Phase: 0.3% Pentafluoropropionic Acid in 43.7mM Ammonium
Formate, pH2.7:0.3% Pentafluoropropionic Acid in
Methanol (45:55)
Flow Rate: 1.0mL/min
Detector: ELSD
1. Streptomycin 2. Amikacin 3. Tobramycin 4. Neomycin
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: A: 0.025M KH2PO4, pH3.0 B: Acetonitrile
Gradient: Time: 0 15 22
%B: 10 40 40
Flow Rate: 1.0mL/min
Detector: UV at 210
10319 10351
Column: Alltech® Alltima HP C18 HiLoad, 5µm, 150 x 4.6mm
(Part No. 87697)
Mobile Phase: 0.1% Phosphoric Acid:Acetonitrile (85:15)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Column: Alltech® Prevail™ Select C18, 5µm, 150 x 4.6mm
(Part No. 99300)
Mobile Phase: Ammonium Hydroxide in Water, pH9.0:Acetonitrile (5:95)
Flow Rate: 1.0mL/min
Detector: ELSD
8711 8764
93969210
1. Tylosin (400ppm) 2. Monensin (180ppm)
1. Neomycin 2. Amoxicillin 3. Vancomycin 4. Cefatrizine 5. Metampicillin 6. Cefotaxime 7. Unknown 8. Bacitracin 9. Pipericillin 10. Bacampicillin 11. Oxacillin
Antibiotics
Antibacterials Antibacterials
Aminoglycoside Antibiotics Antibacterials
Sulfa DrugsTylosin and Monensin
1. Sulfanilamide 2. Sulfamerazine 3. Sulfamethazine 4. Sulfamethizole
12www.discoverysciences.com
®
®
®
1
2
3456 7
80
5
10
15
20
mA
U
0 1 2 3 4 5 Min.0 5 10 15 20 25 30 35 40 45 Min.
1
2
3
pharm
aceuticals
hplc applications
10724
1. Sulfadiazine 2. Sulfathiazole 3. Sulfamerazine 4. Sulfamethazine 5. Sulfamethoxazole 6. Sulfaisoxazole 7. Sulfaquinoxilone 8. Sulfamethoxine
Column: Grace® VisionHT C18-P, 1.5µm, 50 x 2.0mm
(Part No. 5139606)
Mobile Phase: A: 0.1% Formic Acid B: Methanol
Gradient: Time: 0 1.4
%B: 20 100
Flow Rate: 0.4mL/min
Detector: UV at 280nm
10626
1. Amikacin 2. Tobramycin 3. Neomycin
Column: ® A ltima HP HILIC, 5µm, 150 x 4.6mm
(Part No. 86464)
Mobile Phase: Acetonitrile:Methanol:Water (87:11:2)
Flow Rate: 1.0mL/min
Detector: ELSD
10697
Column: C18, 5µm, 250 x 4.6mm
(Part No. 5138810)
Mobile Phase: Monobasic Potassium Phosphate, pH5.0:
Acetonitrile (96:4)
Flow Rate:
Detector: at 230nm
Sulfa DrugsAntibiotics via HILIC
Amoxicillin (U.S.P.)
Antibiotics
0 1 2 3 4 5 6 7 Min.
-500000
0
500000
1000000
1500000
2000000
2500000
3000000
3500000
4000000
4500000
5000000
5500000
6000000
mA
U
Neomycin Extracted from Neo-Cinolone Cream
Column C18 column, 5µm, 250 x 4.6mm
Mobile Phase: 0.3% Pentafluoropropionic Acid in Methanol:
0.3% Pentafluoropropionic Acid in 43.4mM
Ammonium Formate, pH 2.6 (55:45)
Flowrate: 1.0mL/min
Detector: ELSD
1. Neomycin
0 1
1
2 3 4 5 6 7 8 9 Min.
GraceSmart™
1.5mL/min
UV
0.05N
9216
13 www.discoverysciences.com
®®
Alltech
® Alltima ® Alltech
0 1
1
2 3 4 5 6 7 8 9 Min.
ph
arm
ace
utic
als
hplc applications
10680
1. Spectinomycin
Column: Alltech® Alltima HP C18 Amide, 3µm, 50 x 4.6mm
(Part No. 87829)
Mobile Phase: 10mM Ammonium Carbonate:Acetonitrile (98:2)
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Jones Genesis® C18, 4µm, 150 x 4.6mm
(Part No. FM15960E)
Mobile Phase: Acetonitrile:0.1% Trifluoroacetic Acid (31:69)
Flow Rate: 1.0mL/min
Detector: UV at 214nm
Bacitracin SpectinomycinAntibiotics
mA
U
80
100
60
40
20
0
4 62 8 10 12 14 18 Min.160
10760
1. Clarithromycin
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Phosphate Buffer (Dissolve 9.11g Potassium Dihydrogen
Phosphate in 1000mL Water, add 2mL Triethylamine, adjust
with Phosphoric Acid to pH of 5.5):Acetonitrile (60:40)
Flow Rate: 1.0mL/min
Detector: UV at 210nm
Clarithromycin on Chinese Pharmacopeia Assay
mA
U 800
1000
1200
1400
600
400
200
0
5 10 15 25 Min.200
10776
1. Ceftriaxone
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: 0.02mol/L n-Octylamine:Acetonitrile (73:27)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Ceftriaxone Sodium on Chinese Pharmacopeia Assay
14www.discoverysciences.com
®
, , ,
pharm
aceuticals
hplc applications
Bronchodilators
Column: Jones Genesis® C18, 4µm, 150 x 4.6mm
(Part No. FM15960E)
Mobile Phase: Acetonitrile:50mM Aqueous Ammonium Acetate, pH4.7
(12:88)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Benzoate
-Ethoxytheophylline
Xanthines
Clotrimazole
Lotrimin® Topical Assay
Antifungal
10447
Column: Alltech® Alltima C18, 5µm, 150 x 4.6mm (Part No. 88302)
Mobile Phase: A: Water to pH7.0 with Acetic Acid B: Methanol
Gradient: Time: 0 8 10 20 25
%B: 30 50 100 100 30
Flow Rate: 1.0mL/min
Detector: ELSD
Griseofulvin Assay on Chinese Pharmacopeia
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: 0.05mol/L Potassium Dihydrogen Phosphate:
Acetonitrile:Methanol (57:38:5)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Uracil 2. Theobromine 3. Impurity 4. Sodium 5. Theophylline 6. 7. Caffeine
0 2 4 6 8 10 12 14 16 18 20 22 24 Min.
2
1. PEG 2.
mA
U
300
350
400
250
150
200
100
50
0
12.510 17.5155 7.52.5 20 22.5 Min.0
1. Griseofulvin
10772
15 www.discoverysciences.com
®
1
0 1
89
10 11
1213
1
2
2
3
3
4
4,5
5 6
6
7
7
8 Min.
pharm
aceutic
als
hplc applications
Column: Alltech® Alltima Phenyl 3µm, 53 x 7mm Rocket™
(Part No. 43867)
Mobile Phase: 0.05% Trifluoroacetic Acid:Methanol:Acetonitrile
(50:19:31)
Flow Rate: 2.5mL/min
Detector: UV at 230nm
1. 10-Deacetylbaccatin III 2. Baccatin III 3. 7-Xylosyl-10-deacetyltaxol B 4. Taxinine M 5. 7-Xylosyl-10-deacetyltaxol 6. 7-Xylosyl-10-deacetyltaxol C
9221
Taxol® and Related Taxanes in Yew
Taxols
7. 10-Deacetyltaxol 8. 7-Xylosyltaxol 9. Cephalomannine (Taxol® B) 10. 7-Epi-10-deacetyltaxol 11. Taxol®
12. Taxol® C 13. 7-Epitaxol
1. 10-Deacetylbaccatin III 2. Baccatin III 3. 10-Deacetyl-7-xylosyltaxol B 4. Taxinine M 5. 10-Deacetyl-7-xylosyltaxol 6. 10-Deacetyl-7-xylosyltaxol C 7. 10-Deacetyltaxol 8. 7-Xylosyltaxol
Taxanes
9. Cephalomannine 10. 10-Deacetyl-7-epitaxol 11. Taxol®
12. Taxol® C 13. 7-Epitaxol a. 3’-Fluorophenyl-10-deacetyltaxol b. 3’-Fluorophenyltaxol
Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 218MS54)
Mobile Phase: A: 50mM Sodium Acetate, pH6.8 B: Acetonitrile
Gradient: Time: 0 2 32 42 42
%B: 34 34 44 60 100
Flow Rate: 1.0mL/min
Detector: UV at 254nm
mA
U
200
250
300
350
400
450
150
100
50
0
2 4 6 8 10 Min.0
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Water (10:90)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Aciclovir
Aciclovir Assay on Chinese Pharmacopeia
Antiviral
10764
16www.discoverysciences.com
®
mA
U
250
150
200
100
50
0
6 842 10 12 Min.0
mA
U
20
25
30
15
10
5
0
5 10 15 2520 30 35 Min.0
mA
U
120
140
100
60
80
40
20
0
6 842 10 12 Min.0
mA
U
200
250
300
150
100
50
0
2.5 5 7.5 10 12.5 15 17.5 20 22.5 25 Min.0
mA
U
600
700
500
300
400
200
100
0
6 842 1210 14 16 20 Min.180
ph
arm
ace
uti
ca
lshplc applications
Nimodipine Tablets on Chinese Pharmacopeia Assay
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Phosphate Buffer (Dissolve 1.36g Potassium
Dihydrogen Phosphate in 1000mL Water, add 4mL
Triethylamine, adjust with Phosphoric Acid to pH3.5)
(75:25)
Flow Rate: 1.0mL/min
Detector: UV at 253nm
1. Flunarizine 1. Nimodipine
10763
Flunarizine Hydrochloride Capsules on Chinese Pharmacopeia Assay
Calcium Channel Blockers
10761
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Acetonitrile:Water (35:38:27)
Flow Rate: 1.0mL/min
Detector: UV at 237nm
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: 700mL Phosphate Buffer (6.8g Potassium Dihydrogen
Phosphate in 1000mL Water, adjust pH3.0 with
Phosphoric Acid), add Methanol 300mL and Sodium
1-Octanesulfonate 1.30g
Flow Rate: 1.0mL/min
Detector: UV at 275nm
1. Atenolol
Atenolol Assay on Chinese PharmacopeiaBeta Blockers
10777
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Sodium 1-Heptanesulfate solvent (2.0g Sodium
1-Heptanesulfate in 900mL Water, adjust to pH 3.9
with Acetic Acid):Acetonitrile:Methanol (25:6:1)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Famotidine 1. Naloxone
10778
Famotidine Tablets on Chinese Pharmacopeia Assay
Histamine Receptor Antagonists
10779
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Sodium 1-octanesulfonate solvent (1.36g Sodium
1-octanesulfonate, 1.0g Sodium Chloride in 580mL
Water):Methanol:Phosphoric Acid (580:420:1)
Flow Rate: 1.0mL/min
Detector: UV at 229nm
Naloxone Hydrochloride Injection on Chinese Pharmacopeia Assay
Narcotic Analgesic Antagonists
17 www.discoverysciences.com
500000
600000
200000
300000
400000
0
-100000
100000
70 80 906040
12
3
50 100 Min.0
1
2
3
4
5
67
8
9
10
11
20 40 60 Min.
1
2
3
4
0 2.5 5 7.5 10 12.5 15 17.5 20 22.5 25 Min.
hplc applicationsn
utra
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utic
als
| vita
min
s
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile:Water:Isopropyl Alcohol (80:5:15)
B: Acetonitrile:Water:Isopropyl Alcohol (67:21:12)
Gradient: Time: 0 28 35 45 50 51 57
%B: 25 85 80 75 90 100 25
Flow Rate: 0.8mL/min
Detector: ELSD
1. Rh2 2. Rh1 3. Rg2 4. Rg3 5. Rg1 6. Rf
7. Re 8. Rd 9. Rc 10. Rb2 11. Rb1
10239 10287
10288
1. Bilobalide 2. Ginkgolide J 3. Ginkgolide C 4. Ginkgolide A 5. Ginkgolide B
1. Bilobalide 2. Ginkgolide J 3. Ginkgolide C 4. Ginkgolide A 5. Ginkgolide B
10142
1. Quercetin 2. Kaempferol 3. Isorhamnetin
Column: Alltech® Alltima C18, 3µm, 100 x 4.6mm
(Part No. 81382)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.1% Trifluoroacetic Acid in Methanol
Gradient: Time: 0 15
%B: 10 55
Flow Rate: 1.3mL/min
Detector: ELSD
Column: Alltech® Platinum™ C18, 5µm, 250 x 4.6mm
(Part No. 32064)
Mobile Phase: A: 0.05% Trifluoroacetic Acid in Water:Methanol (95:5)
B: 0.05% Trifluoroacetic Acid in Methanol
Gradient: Time: 0 20
%B: 10 90
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™
(Part No. 50605)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.1% Trifluoroacetic Acid in Methanol
Gradient: Time: 0 5
%B: 10 45
Flow Rate: 5.0mL/min
Detector: ELSD
10742
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Tetrahydrofuran:Water (25:10:65)
Flow Rate: 1.0mL/min
Detector: ELSD
1. Ginkgolide C 2. Bilobalide 3. Ginkgolide A 4. Ginkgolide B
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: A: Water B: Acetonitrile
Gradient: Time: 0 35 55 70 100
%B: 19 19 29 29 40
Flow Rate: 1.0mL/min
Detector: ELSD
10765
Ginseng
Ginsenosides in Korean Ginseng
Ginkgo Biloba
Ginkgo Biloba Flavonol Glycoside Standards Ginkgo Biloba Terpene Lactone Standards
Terpene Lactones High ThroughputFolium Ginkgo
Ginsenoside
1. Rg1 2. Re 3. Rb1
18www.discoverysciences.com
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®
1
2
3
54
0 5 10 15 20 25 30 35 40 45 Min.
0 5
1
2
3
4
5 6
10 15 Min.
hplc applicationsn
utr
ace
uti
ca
ls | v
ita
min
s
10315
Column: Alltech® Alltima C18, 5µm, 150 x 10mm (Part No. 81102)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.1% Trifluoroacetic Acid in Methanol
Gradient: Time: 0 45
%B: 22 32
Flow Rate: 5.0mL/min
Detector: ELSD
1. Bilobalide
2. Ginkgolide J
3. Ginkgolide C
4. Ginkgolide A
5. Ginkgolide B
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: A: 0.025M KH2PO4, pH2.5 B: Acetonitrile
Gradient: Time: 0 15 25
%B: 20 40 40
Flow Rate: 1.0mL/min
Detector: UV at 280
1. Myricetin 2. Quercetin 3. Naringenin 4. Hesperetin 5. Apigenin 6. Kaempferol
Column: Alltech® Alltima C18 3µm, 53 x 7mm Rocket™
(Part No. 50605)
Mobile Phase: 0.1% Trifluoroacetic Acid in Water:
0.1% Trifluoroacetic Acid in Acetonitrile (65:35)
Flow Rate: 2.5mL/min
Detector: ELSD
1. Myricetin 2. Quercetin 3. Apigenin 4. Kaempferol
Column: Alltech® Alltima C18, 3µm, 100 x 4.6mm (Part No. 81382)
Mobile Phase: A: 0.05M K2HPO4, pH2.5 B: Acetonitrile
Gradient: Time: 0 15
%B: 20 40
Flow Rate: 1.0mL/min
Detector: UV at 280nm
9107 9110
1. Protocatechuic Acid 2. Chlorogenic Acid 3. Caffeic Acid
Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™
(Part No. 50605)
Mobile Phase: 0.01M K2HPO4, pH2.6:Acetonitrile (80:20)
Flow Rate: 3.5mL/min
Detector: UV at 330nm
1. Pseudohypericin 2. Hypericin
Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™
(Part No. 50605)
Mobile Phase: 0.2% Phosphoric Acid:Methanol (5:95)
Flow Rate: 4.0mL/min
Detector: Vis at 585nm
9397
9271 8887
1. Naringin 2. Neohesperidin 3. Myricetin 4. Quercetin 5. Naringenin 6. Kaempferol
Liquid Ginkgo Biloba Extract Prep
Flavonoids
Echinacea
Polyphenolic Flavonoids
Flavonoid Mix Flavonoid Mix
St. John’s Wort
Phenolic AcidsHypericins
Ginkgo Biloba
19 www.discoverysciences.com
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® ®
® ®
2
1
0 2 4 6 8 10 12 14 Min.
0 2 4 6 8 10 12 14 16 18 20 Min. 0 1 2 3 4 5 6 7 8 9 10 Min.
0 2 4 6 8 10 12 14 16 Min.
0 2 4 6 8 10 12 Min.
hplc applicationsn
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s
10747
1074610745
10744
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Water:Acetic Acid (87:13:0.1)
Flow Rate: 1.0mL/min
Detector: ELSD
10743
1. Peimine 2. Peininine
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Water:Acetic Acid (85:15:0.2)
Flow Rate: 1.0mL/min
Detector: ELSD
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm
(Part No. 5138810)
Mobile Phase: Acetonitrile:Water (35:65)
Flow Rate: 1.0mL/min
Detector: ELSD
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Acetonitrile:Water:Diethylamine (70:30:0.3)
Flow Rate: 1.0mL/min
Detector: ELSD
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Water (95:5)
Flow Rate: 1.0mL/min
Detector: ELSD
1. Liguidanbaric Acid 1. Momordin lc
1. Sarsasapogenin
1. Astragaloside IV
Chinese Pharmacopeia
Fructus Liquidambaris Fructus Kochiae
Bulbus Fritillariae Thunbergii Rhizoma Anemarrhenae
Radix Astragali
20www.discoverysciences.com
mA
U
50
20
30
40
10
0
25 30 35 4015 205 10 45 Min.0
mA
U
500
200
300
400
0
100
15 20 25100 5 30 Min.
0 2
12
3
4 6 8 10 12 14 16 18 20 Min.
0 1
1
2
2
3
3
4
5
6
7 8
4
5 Min.
hplc applicationsn
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min
s
Column: Alltech® Alltima C18, 5µm, 150 x 22mm (Part No. 81106)
Mobile Phase: 0.05% Trifluoroacetic Acid:Acetonitrile (87:13)
Flow Rate: 23mL/min
Detector: ELSD
1. Caffeine 2. Epicatechin 3. Epigallocatechin Gallate
Column: Alltech® Alltima C18, 5µm, 150 x 4.6mm (Part No. 88052)
Mobile Phase: 0.05% Trifluoroacetic Acid:Acetonitrile (87:13)
Flow Rate: 1.0mL/min
Detector: ELSD
10139
1. Caffeine 2. Epicatechin 3. Epigallocatechin Gallate
Column: Alltech® Platinum™ EPS C18,
1.5µm, 33 x 7mm Rocket™ (Part No. 50577)
Mobile Phase: 0.05% Trifluoroacetic Acid:Acetonitrile (87:13)
Flow Rate: 2.5mL/min
Detector: UV at 210nm
1. Gallic Acid 2. Epigallocatechin 3. Catechin 4. Epicatechin 5. Epigallocatechin Gallate 6. Caffeine 7. Gallocatechin 8. Epicatechin Gallate
10140
8825
Green Tea
Green Tea Extract - PreparativeGreen Tea Extract - Analytical
Catechins
1. Gallic Acid 2. Gallocatechin 3. Epigallocatechin 4. Catechin 5. Epicatechin 6. Epigallocatechin Gallate 7. Gallocatechin Gallate 8. Epicatechin Gallate 9. Catechin Gallate
Catechins and Gallic Acid in Green Tea
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:0.1% Citric Acid (25:75)
Flow Rate: 1.0mL/min
Detector: UV at 250nm
10773
1. Puerarin
Puerarin
Puerarin
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Water (75:25)
Flow Rate: 1.0mL/min
Detector: UV at 270nm
10769
1. Tanshinone II
Tanshinone II
Tanshinone II
Column: Vydac® Denali®, 5µm, 150 x 4.6mm (Part No. 238DE5415)
Mobile Phase: A: Acetonitrile:25mM Phosphoric Acid (10:90)
B: Acetonitrile:25mM Phosphoric Acid (90:10)
Gradient: Time: 0 3 22
B%: 0 0 15
Flow Rate: 1.0mL/min
Detector: UV at 280nm
21 www.discoverysciences.com
® ®
Min.
0 1 2 3 4 5 6 7 8 9 10 Min.
1
2
3
2
3
4
1
5
6
7
0 2 4 6 8 10 12 14 Min.
0 2 4 6 8 10 12 14 16 Min.
5 6
71
2
43
0 2 4 6 8 10 12 14 Min.
1
2
3
4
5
6
7
hplc applicationsn
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min
s
1. Thiamine 2. Ascorbic Acid 3. Nicotinic Acid 4. Niacinamide 5. Pantothenic Acid 6. Riboflavin 7. Cyanocobalamin 8. Biotin 9. Vitamin A 10. -Tocopherol 11. -Tocopherol 12. -Tocopherol 13. Vitamin D2 14. Vitamin D3 15. Vitamin K
9545
1. Vitamin A (100ng on column) 2. Vitamin E (160ng on column) 3. Vitamin K1 (130ng on column)
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: Acetonitrile:Methanol (75:25)
Flow Rate: 1.5mL/min
Detector: UV at 220nm
1. Vitamin A 2. -Tocopherol 3. -Tocopherol 4. -Tocopherol (Vitamin E) 5. Vitamin D2 6. Vitamin D3 7. Vitamin K
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: 0.025M KH2PO4, pH3.0:Acetonitrile (97:3)
Flow Rate: 1.0mL/min
Detector: UV at 212
1. Thiamine 2. Ascorbic Acid 3. Unknown 4. Fumaric Acid 5. Pyridoxine 6. Niacinamide 7. Pantothenic Acid
1. Ascorbic Acid 2. Nicotinic Acid 3. Pyridoxamine 4. Niacinamide 5. Folic Acid 6. Riboflavin 7. Vitamin B12
Column: Alltech® Platinum™ C18, 5µm, 150 x 4.6mm
(Part No. 32043)
Mobile Phase: A: 0.1M Ammonium Phosphate, pH4.5
B: Acetonitrile
Gradient: Time: 0 4 15 18
B%: 5 5 40 40
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Column: Alltech® Alltima C18, 3µm, 150 x 1mm (Part No. 88384)
Mobile Phase: Methanol:Chloroform (90:10)
Flow Rate: 0.05mL/min
Detector: ELSD
9390
81549456
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: A: Water pH3.2 w/Formic Acid
B: Acetonitrile:Methanol (83:17)
Gradient: Time: 0 3 10 10.1 30
%B: 0 0 45 100 100
Flow Rate: 1.5mL/min
Detector: ELSD
10279
1. Ascorbic Acid 2. Nicotinic Acid 3. Pyridoxamine 4. Niacinamide 5. Folic Acid 6. Vitamin B12 7. Riboflavin
Column: GraceSmart™ C18, 5µm, 150 x 4.6mm (Part No. 5138812)
Mobile Phase: A: 0.1M Ammonium Dihydrogen Orthophosphate
Buffer, pH4.5
B: Acetonitrile
Gradient: Time: 0 4 10 15
B%: 5 5 40 40
Flow Rate: 1.0mL/min
Detector: UV at 254nm
10733
Fat-Soluble Vitamins
Water- and Fat-Soluble Vitamins
Fat-Soluble Vitamins and Tocopherols
Water-Soluble VitaminsWater-Soluble Vitamins
from Multi-Vitamin Tablet
Water-Soluble Vitamins
Vitamins
22www.discoverysciences.com
®
Column: VYDAC 201TP54 (C18, 5 µm, 4.6 mm i.d. x 250 mm)
hplc applicationsn
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s
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: Acetonitrile:Methanol (75:25)
Flow Rate: 1.5mL/min
Detector: ELSD
1. -Tocopherol 2. -Tocopherol 3. -Tocopherol
9458
Vitamin E Soft Gel Tablet
1. All-trans-Retinol 2. Tocol 3. -Tocopherol 4. -Tocopherol 5. Lutein 6. Zeaxanthin 7. Cryptoxanthin 8. -Carotene 9. All-trans- -Carotene 10. cis- -Carotene
Determination of Retinol, -Tocopherol, and -Carotene in Serum
Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 201TP54)
Mobile Phase: A: Buffer:Methanol:n-Butanol (15:75:10)
B: Buffer:Methanol:n-Butanol (2:88:10)
Buffer:Aqueous 0.02M Ammonium Acetate, pH3.5
Gradient: Time: 0 3 18 32
%B: 0 0 100 100
Flow Rate: 1.5mL/min
Detector: Absorbance, programmed wavelength
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: Methanol:Ethanol (50:50)
Flow Rate: 1.0mL/min
Detector: UV at 275nm
10771
Coenzyme Q10
mA
U
80
100
60
40
20
0
15 20105 25 30 35 45 Min.400
1. Coenzyme Q10
Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 201TP54)
Mobile Phase: Methanol:Chloroform (94:6)
–Carotenes
Detector: UV Flow Rate: 1.0mL/min
1. -Carotene 2. cis- -Carotene 3. -Carotene 4. 13-cis- -Carotene 5. 9-cis- -Carotene
1. Vitamin D2
2. Vitamin D3
Column: Adsorbosphere™ UHS C18, 5µm, 4.6 x 150mm
(Part No. 288118)
Mobile Phase: 80% Acetonitrile 20% Methanol
Flow Rate: 1.5mL/min
Detector: UV at 254nm
Column Temp: Ambient
Vitamins D2 & D3
5401
Vitamins
23 www.discoverysciences.com
0 10 20 30 40 Min.
1
2
45
67
8
1
1
UV
RI
ELSD
0 1 2 3 4 5 Min.
Column: Alltech® Alltima C18, 5µm, 250 x 4.6mm (Part No. 88056)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 40
%B: 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
1. LGG 2. LGL 3. LLL 4. OGL 5. PGL 6. OLL 7. OGO + PLL 8. OOO
10047
Evening Primrose
Triglycerides
10412
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 150 x 4.6mm
(Part No. 35102)
Mobile Phase: Acetonitrile:Water (65:35)
Flow Rate: 1.0mL/min
Detector: ELSD
10413
10411
Glucosamine
1. Glucosamine
Column: Alltech® Prevail™ C18, 5µm, 250 x 4.6mm (Part No. 99210)
Mobile Phase: A: 1% Formic Acid B: Acetonitrile
Gradient: Time: 0 60 65 67 80
%B: 20 100 100 20 20
Flow Rate: 1.0mL/min
Detector: ELSD
1. Cimiracemoside A 2. Actein 3. 27-deoxyactein
9496
Black Cohosh Extract
Black Cohosh
Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™
(Part No. 50605)
Mobile Phase: 0.02M KH2PO4, pH2.6:Methanol (52:48)
Flow Rate: 3.0mL/min
Detector: UV at 235nm
1. Hydrastine 2. Berberine
9124
Alkaloids
Goldenseal Root
Glucosamine
Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™
(Part No. 50605)
Mobile Phase: 0.05% Trifluoroacetic Acid:Methanol:Dioxane (75:23:2)
Flow Rate: 3.0mL/min
Detector: UV at 285nm
9191
Iridoid Compounds
Devil’s Claw
1. 8-para-Coumaroyl Harpagoside 2. Harpagoside
0 1
1
2
2 3 4 5 6 Min.
Column:
Mobile Phase: Dioxane:ACN:0.05% TFA (3:64:33)Flowrate: 3mL/minDetector: UV at 220nm
1. Acetoxyvalerenic Acid2. Valerenic Acid
Valerian
Valerian
9160
® Alltima C18, 3µm, 53 x 7mm Rocket™
(Part No. 50605) Alltech
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Carbohydrates
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0 10 20 30 40 50 60 Min.
1
2
3
45 6
7
8
910
11
1213
14
15
16
17
18
1920
21
22
0 1 2 3 4 Min.
1
2
3
Column: Alltech® Prevail™ Carbohydrate ES,
5µm, 53 x 7mm Rocket™ (Part No. 35104)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 4
%B: 32 50
Flow Rate: 2.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Carbohydrate ES, 5µm,
250 x 4.6mm (Part No. 35101)
Mobile Phase: A: Acetonitrile B: 0.04% Ammonium Hydroxide in Water
Gradient: Time: 0 25 40 60
%B: 17 27 45 55
Flow Rate: 1.0mL/min
Detector: ELSD
9420 1. Dextrose 2. Maltose 3. Maltotriose 4. Maltotetraose 5. Maltopentaose 6. Maltohexaose 7. Maltoheptaose 8. Maltooctaose
10009
Column: Alltech® Prevail™ Carbohydrate ES, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 50
%B: 20 35
Flow Rate: 1.0mL/min
Detector: ELSD
1. iso-Erythritol 2. Fructose 3. Sorbitol 4. Mannitol 5. Glucose 6. Inositol 7. Sucrose 8. Maltitol 9. Maltose
Column: Alltech® Prevail™ Carbohydrate ES, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile:Acetone (75:25)
B: Water
Gradient: Time: 0 18
%B: 24 50
Flow Rate: 1.0mL/min
Detector: ELSD
10. Raffinose 11. DP3 12. DP4 13. DP5 14. DP6 15. DP7 16. DP8 17. DP9 18. DP10 19. DP11 20. DP12 21. DP13 22. DP14
Column: Alltech® Prevail™ Carbohydrate ES,
5µm, 53 x 7mm Rocket™ (Part No. 35104)
Mobile Phase: Acetonitrile:Water (75:25)
Flow Rate: 2.0mL/min
Column Temp: 30°C
Detector: ELSD
10277
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 15
%B: 35 50
Flow Rate: 1.0mL/min
Detector: ELSD
1. Fructose 2. Glucose 3. Sucrose 4. Maltose 5. Maltotriose 6. Maltotetraose 7. Maltopentaose
1. Fructose 2. Glucose 3. Sucrose 4. Maltose 5. Maltotriose 6. Maltotetraose 7. Maltopentaose
9350 9360
1. Fructose 2. Glucose 3. Sucrose
1. Dextrose 2. Maltose 3. Maltotriose 4. Maltotetraose 5. Maltopentaose
6. Maltohexaose 7. Maltoheptaose 8. Maltooctaose 9. Maltononaose 10. Maltodecaose 11. Maltoundecaose 12. Maltododecaose 13. Maltotridecaose
9354
Corn Syrup SolidsCorn Syrup Oligomers
Mono-, Di-, Tri- Saccharides Sugars
Beer Standards
Unmodified Mobile Phase
Beer Standards
Modified Mobile Phase
0 5 10 15 20 25 30 35 40 45 Min.
1
23
4
0
1
2
3 5
7
8 9
10
4
6
10 20 30 40 50 Min.
0 1 2 3 Min.
1
2
3
4
5
Column: Alltech® Prevail™ Carbohydrate ES, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: Acetonitrile:Water (70:30)
Flow Rate: 1.0mL/min
Detector: ELSD
1. iso-Erythritol 2. Xylitol 3. Mannitol 4. Maltitol
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 50
%B: 20 35
Flow Rate: 1.0mL/min
Detector: ELSD
9351
9474
Column: Alltech® IOA-1000, 9mm, 300 x 7.8mm (Part No. 9646)
Mobile Phase: 0.005N Sulfuric Acid
Flow Rate: 0.3mL/min
Column Temp: 60°C
Detector: RI
5411
9446
9352
Column: Alltech® Prevail™ C18, 3µm, 50 x 2.1mm (Part No. 43818)
Mobile Phase: Acetonitrile:0.05M Ammonium Acetate, pH5.4, (20:80)
Flow Rate: 0.3mL/min
Detector: UV at 230
1. Ascorbic Acid 2. Saccharin 3. Benzoic Acid 4. Aspartame 5. Vanillin
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 60
%B: 17 35
Flow Rate: 1.0mL/min
Detector: ELSD
1. Fructose 2. Sorbitol 3. Glucose 4. Maltose
1. Fructose 2. Glucose 3. Maltose 4. Maltotriose 5. Maltopentaose 6. Maltohexaose 7. Maltoheptaose
1. Citric Acid 2. Tartaric Acid 3. Glucose 4. Malic Acid 5. Fructose 6. Acetic Acid 7. Glycerol 8. Lactic Acid 9. Methanol 10. Ethanol
Sugar Alcohols Wine Analysis - 2000 Sipaska Rosé
Wine Analysis by Ion Exclusion
Beverage AdditivesImported Ale
Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 218TP54)
Gradient: 7 to 10% Methanol over 60 min.,
then to 25% Methanol at 90 min
Flow Rate: 1.0mL/min
Detector: ELSD
Partial Digest: Complex Polysaccharide
from Plant Cell Wall
Carbohydrates
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0 2 4 6 8 10 12 14 16 18 Min.
2 4
53
1
0 5 10 15 20 25 30 35 40 Min.
12
3 4
0 5 10 15 20 25 30 35 40 Min.
1
87
65
4
32
0 5 10 15 20 25 Min.
2
1
3
0 5 10 15 20 25 30 35 40 Min.
2
1
0 1 2 3 4 5 6 7 8 9 Min.
1
2 3
10531 1. Sucralose 2. Glucose 3. Maltose 4. Maltodextrins
10530
1. Sucralose 2. Unknown 1 3. Fructose 4. Glucose 5. Unknown 2 6. Sucrose 7. Maltose 8. Maltodextrins
10529
1. Fructose 2. Glucose 3. Sucrose
10579
1. Sucrose 2. Lactose
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: Acetonitrile:Water (75:25)
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Carbohydrate ES, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 30
%B: 20 29
Flow Rate: 1.0mL/min
Detector: ELSD
1. Fructose 2. Mannitol 3. Glucose 4. Sucrose 5. Unknown
10585
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 5 10 15 25
%B: 20 20 30 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 5 10 15 25
%B: 20 20 30 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 5 10 15 25
%B: 20 20 30 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
10584
1. Sucralose 2. Rhamnose 3. Cellobiose
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 150 x 4.6mm
(Part No. 35102)
Mobile Phase: Acetonitrile:Water (70:30)
Flow Rate: 1.0mL/min
Detector: ELSD
Spiked UrineChromatogram courtesy of Rosemary Rose,
Monash University, Dept. of Gastroenterology
Conquistador® Tequila Splenda® Sweetener
Pepsi One® Cola Infant Formula
Flavor Sample SolutionChromatogram courtesy of Charles Stewart, Firmenich Inc.
Carbohydrates
27 www.discoverysciences.com
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Min. Min.
Min. Min.
Min. Min.
0 5 10 15 20 25 30 35 40 Min.
1
2
3
4
5 6
0 5 10 15 20 25 30 35 40 Min.
4
3
1 2 5
45
3
21
1/3 Less Sugar
Regular
0 5 10 15 20 25 30 35 40 Min.
1
3
2
4
Kellogg's Frosted Flakes® Cereal
Regular vs. 1/3 less sugar
1. Sucralose 2. Fructose 3. Sorbitol 4. Glucose 5. Sucrose 6. Maltose
10526
10557
1. Fructose 2. Glucose 3. Sucrose 4. Maltose 5. Oligosaccharides
1. Fructose 2. Glucose 3. Sucrose 4. Unknown
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 5 10 15 25
%B: 20 20 30 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 5 10 15 25
%B: 20 20 30 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 5 10 15 25
%B: 20 20 30 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm
(Part No. 35101)
Mobile Phase: A: Acetonitrile B: Water
Gradient: Time: 0 5 10 15 25
%B: 15 15 25 25 50
Flow Rate: 1.0mL/min
Detector: ELSD
1. Fructose 2. Glucose 3. Sucrose 4. Maltose
10556
10527
10528
Carbohydrates
Robitussin® Flu Syrup
Tobacco
Cough Syrup
Vegetable Fermentation Broth Standards
1. iso-erythritol
2. Fructose
3. Sorbitol
4. Mannitol
5. Glucose
6. Inositol
7. Sucrose
8. Maltitol
9. Maltose
10. Raffinose
11. MaltotrioseColumn: Prevail™ Carbohydrate ES Column, 250 x 4.6mm, (Part No. 35101)Mobile Phase: A: Acetonitrile B: 0.04% NH
4OH
Gradient: Time: 0 60 %B: 17 35Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient
0 10 20 30 40 50 Min.
12
3
4
5
6
7
8
9
10
11
10004
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Sugar Alcohols
Column: Alltech® Prevail™ Carbohydrate ES column, 5µm,
250 x 4.6mm, (Part No. 35101)
Mobile Phase: Acetonitrile:Water (79:21)
Flow Rate: 1.0 mL/min
Column Temp: 30°C
10627
10635
18 Min.
2 31
ELSD
RI
Solvent
Front
0 2 4 6 8 10 12 14 16
32
1
1. Xylitol (200ppm)
2. Sorbitol (200ppm)
3. Dulcitol (200ppm)
Orange Juice Profiles
9532
9531
9530
Column: Prevail™ Carbohydrate ES Column, 5µm, 53 x 7.0mm Rocket™ (Part No. 35104) Mobile Phase: Acetonitrile:Water (75:25)Flow Rate: 1.3mL/min Detector: ELSDColumn Temp: Ambient
0 1 2 3 4 5 6 7 8 Min.
1
2
Orange Drink
1
2
3
Dominick’s® 100%
1
2
3
Minute Maid® 100%
1. Fructose
2. Glucose
3. Sucrose
Carbohydrate Analysis with ELSD Improves Baseline Stability and Detection Sensitivity
Compared to RI and Low-Wavelength UV
Column: Prevail™ Carbohydrate ES Column, 250 x 4.6mmMobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 20 25 %B: 20 50 50Flow Rate: 1.0mL/minColumn Temp: Ambient
9355
9380
20 4 6 8 10 12 14 16 18 20 22 Min.
12
34
5 6UV at 195nm
1 2
3
4
56
ELSDRI
0 1 2 3 4 Min.
1
23
1 2 3
Solvent
Front
ELSD
Column: Prevail™ Carbohydrate ES Column, 5µm, 53 x 7.0mm Rocket™ (Part No. 35104) Mobile Phase: Acetonitrile:Water (75:25)Flow Rate: 2.0mL/min Column Temp: 30°C
10278
10277
1. Mannitol
2. Fructose
3. Glucose
4. Sucrose
5. Raffinose
6. Stachyose
1. Fructose
2. Glucose
3. Sucrose
30
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3
Wine - Concord Grape
Column: Prevail™ Carbohydrate ES Column, 5µm, 250 x 4.6mm (Part No. 35101)Mobile Phase: A: Acetonitrile B:WaterGradient: Time: 0 60 %B: 17 35Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient
1. Sorbitol
2. Fructose
3. Maltose
4. Inositol
10 15 20 25 30 35 40 45 Min.
12
4
Low Level Detection - 10ng on Column
1. Fructose
2. Sucrose
3. Raffinose
Column: Prevail™ Carbohydrate ES Column, 5µm, 150 x 4.6mm, (Part No. 35102) Mobile Phase: A: Water B: Acetonitrile Gradient: Time: 0 10 11 14 %B: 80 50 80 80Flow Rate: 1.0 mL/min Detector: ELSDColumn Temp: Ambient
0 1 2 3 4 5 6 7 8 9 10 Min.
2
1 3
Column: 250 x 4.6mm as identifiedMobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 15 %B: 35 50Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient
Enhance Detector Sensitivity With Stable Gradient Baselines
Carbohydrate ES
1. Dextrose
2. Maltose
3. Maltotriose
4. Maltotetraose
5. Maltopentaose
6. Maltohexaose
7. Maltoheptaose
8. Maltooctaose
9. Maltononaose
10. Maltodecaose
11. Maltoundecaose
12. Maltododecaose
13. Maltotridecaose
14. Maltotetradecaose
15. Maltopentadecaose
12 3
47
5 6
0 5 10 15 20 25 Min.
8 9
1
2
3
4 56
71011 1312
18
12
1314
15
19 20 21 22 23 24 Min.
9354
9379
9486
Prevail™
Column
Silica Based Amino Column
Carbohydrates
9476 10498
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Clover Honey
Column: Prevail™ Carbohydrate ES Column, 5µm, 250 x 4.6mm, (Part No. 35101)Mobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 15
%B: 25 40
Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient
1. Fructose
2. Glucose
3. Sucrose
0 2
2
1
4
3
6 8 10 12 14 16 18 20 Min.
1. Glucose (DP1)
2. DP3 (1,6-alpha; 1,4-alpha)
3. DP6 (1,6-alpha; 1,4-alpha)
4. DP13
5. DP18
Column: Prevail™ Carbohydrate ES Column, 5µm 250 x 4.6mm (Part No. 35101)Mobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 50 60 %B: 35 60 60Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient
Glucooligosaccharides in Maltodextrin
0 6 12 18 24 30 36 Min.
1
23
4 5
21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 Min.
4
5
9535
0 5
1
2
3
4
5
10 15 Min.
1. Unknown 2. Sucrose 3. Glucose 4. Fructose 5. Sorbitol
Column: Alltech® Carbohydrate Cation, 10µm, 6.5 x 300mm
(Part No. 70057)
Mobile Phase: Water
Flow Rate: 0.5mL/min
Column Temp: 90°C
Detector: Alltech ELSD
100% Pure Red Delicious Apple Juice
(a 1:250 Dilution)
Carbohydrates
9534
9365 8965
Column: Prevail™ Carbohydrate ES Column, 5µm, 250 x 4.6mm, (Part No. 35101)Mobile Phase: A: Acetonitrile B: WaterGradient: Step (5 min. hold at 40% B, 5% B steps every 5 min. to 70% B)Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient
Partially Hydrolyzed Dextran
0 5 10 15 20 25 30 35 40 Min.
0 5 10 15 20 25 Min.
1
2
3
45
67
8
910
11
0 1 2 3 4 5 6 7 8 Min.
1
2
3
4
5
0 2 4 6 8 10 Min.
Column: Alltech® Prevail™ Organic Acid, 5µm, 150 x 4.6mm
(Part No. 88640)
Mobile Phase: 0.025M KH2PO4, pH2.5
Flow Rate: 1.5mL/min
Detector: UV at 210nm
1. Oxalic Acid 2. Tartaric Acid 3. Formic Acid 4. Malic Acid 5. Lactic Acid 6. Acetic Acid 7. Diglycolic Acid 8. Maleic Acid 9. Citric Acid 10. Succinic Acid 11. Fumaric Acid
9384
Column: Alltech® Prevail™ Organic Acid, 5µm, 150 x 4.6mm
(Part No. 88640)
Mobile Phase: 0.025M KH2PO4, pH2.5
Flow Rate: 1.0mL/min
Detector: UV at 210nm
1. Tartaric Acid 2. Malic Acid 3. Lactic Acid 4. Citric Acid 5. Fumaric Acid
Organic AcidsOrganic Acid Mix Rye Bread Extract
9484
Carbohydrates
9540
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2
3
4
5
6
7
8 9 10
1
UV
0
3
4
5
6
7
8
9
10
2 4 6 8 10 12 14 16 18 Min.
ELSD
Column: ® Alltima HP C18 HiLoad, 3µm, 150 x 4.6mm
(Part No. 87686)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.1% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 10 20
%B: 80 100 100
Flow Rate: 1.0mL/min
7. Linoleic Acid 8. Palmitic Acid 9. Oleic Acid 10. Stearic Acid
1. Caproic Acid 2. Caprylic Acid 3. Capric Acid 4. Lauric Acid 5. Linolenic Acid 6. Myristic Acid
Organic Acids
Fatty Acids Using UV and ELSD in Tandem
10430
Column: Alltech® Prevail™ Organic Acid, 5µm, 150 x 4.6mm
(Part No. 88640)
Mobile Phase: Acetonitrile:Methanol (75:25)
Flow Rate: 1.0mL/min
Detector: ELSD
1. Lauric Acid 2. Myristic Acid 3. Palmitic Acid 4. Stearic Acid 5. Arachidic Acid 6. Behenic Acid
9539
Underivatized Fatty Acids
Column: Alltech® Platinum™ EPS C8, 5µm, 150 x 4.6mm
(Part No. 32420)
Mobile Phase: 0.025M KH2PO4, pH2.5:Methanol (97:3)
Flow Rate: 0.7mL/min
Detector: UV at 220nm
1. Tartaric Acid 2. Malic Acid 3. Lactic Acid 4. Diglycolic Acid 5. Maleic Acid 6. Succinic Acid 7. Fumaric Acid
Organic Acid Mix
8244
Column:
Mobile Phase: THF:CH 3 CN:0.1% H 3PO4 (25:55:20)
Flowrate: 1.5mL/min
Detector: UV at 220nm
1. Capric Acid (C10)
2. Lauric Acid (C12)
3. Myristic Acid (C14)
4. Palmitic Acid (C16)
5. Stearic Acid (C18)
Fatty Acids
0 2 4 6 8 10 12 Min.
1
2
3
4
5
1
2
3
4
5
6
7
8
9
10
11
12
0 10 20 30 Min.
1. Oxalic Acid 2. Citric Acid 3. Shikimic Acid 4. Fumaric Acid 5. Butyric Acid 6. Homoprotocatechuic Acid 7. Gallic Acid 8. Protocatechuic Acid 9. Gentisic Acid 10. p-Hydroxybenzoic Acid 11. Benzoic Acid 12. Salicylic Acid
7067
Short-Chain Aromatic Carboxylic Acids
Column: Alltech® OA-2000, 6.5µm, 100 x 6.5mm (Part No. 9048)
Mobile Phase: 0.01N Sulfuric Acid
Column Temp: 45°C
Flow Rate: 0.8mL/min
Detector: UV at 228nm
8060
1. Capric Acid 2. Lauric Acid 3. Myristic Acid 4. Palmitic Acid 5. Stearic Acid
9459
Long Chain Fatty Acids
0 1 2 3 4 Min.
1
2
3
4
5
Column: Alltech® Prevail™ C18, 3µm, 53 x 7mm Rocket™
(Part No. 99279)
Mobile Phase: Acetonitrile:Methanol (75:25)
Flow Rate: 3.5mL/min
Detector: ELSD
Alltech
® Alltima C18, 5µm, 150 x 4.6mm Alltech ®
®
1
2 3
4
5
0 1 2 3 4 5 6 7 Min.
14
13
12
11
10
98
4
56
7123
0 5 10 15 20 25 30 Min.
6
4
3
7
8
52
1
0 5 10 15 20 Min.
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)
Mobile Phase: 0.025M KH2PO4, pH2.5:Methanol (97:3)
Flow Rate: 0.7mL/min
Detector: UV at 220
1. Tartaric Acid 2. Malic Acid 3. Lactic Acid 4. Maleic Acid 5. Fumaric Acid
10705
1. LLO 2. OOLn 3. PLnO 4. LOO 5. PoOO 6. PLO 7. PoOP 8. OOO 9. POO+SOL 10. POP 11. SOO 12. POS+SLS 13. PPS 14. C17
10561
Column: Alltech® Prevail™ Silica, 3µm, 150 x 3.0mm (Part No. 99341)
Mobile Phase: A: Chloroform; B: Methanol; C: Triethylamine buffer pH3
Gradient: Time: 0 16 17
%B: 12 60 12
%C: 0.5 12 0.5
Flow Rate: 0.5mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm
(Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Olive Oil
1. LACCER 2. PA 3. PE 4. PI 5. PS 6. PC 7. SM1 8. SM2
10576
Acid Whey
Organic AcidsOrganic Acids
0 2 4 6 8 10 12 14 16 Min.
12
3
4
5
6
7
89
1. LLO 2. LLP 3. OOL 4. POL 5. PPL 6. OOO 7. OOP 8. PPO 9. OOS
Column: Alltech® Alltima C18, 3µm, 150 x 4.6mm
(Part No. 81387)
Mobile Phase: A: Dichloromethane B: Acetonitrile
Gradient: Time: 0 10 18 20
%B: 70 55 70 70
Flow Rate: 1.5mL/min
Detector: ELSD
9515
Oils / LipidsOlive Oil
70 Min.0 10 20 30 40 50 60
ELSD
UV at 254nm
10656
Column: Alltech® Alltima
(Part No. 43852)
Mobile Phase: A: 0.1% Formic Acid B: Methanol
Gradient: Time: 0 40
%B: 60 100
Flow Rate: 0.25mL/min
Column Temp: 30°C
Plant Oil Extract
10657
34www.discoverysciences.com
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® C18, 3µm, 2.1 x 150mm®
®
Oils / Lipids
35 www.discoverysciences.com
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0 2 4 6 8 10 12 14 16 Min.
1
2
3
4
5
6
7
8
910
0 2 4 6 8 10 12 14 16 Min.
1
2
3
4
56
0 5 10 15 20 25 30 35 Min.
1
2
3
4
5
6
78
9
10
0 10 20 30 40 Min.
1
2
3
45
67
8
13
4
56
7
8
910
11
12
14
132
0 5 10 15 20 25 30 35 Min.
64
3
5
2
1
0 5 10 15 20 Min.
Column: ® Alltima C18, 3µm, 150 x 4.6mm (Part No. 81387)
Mobile Phase: A: Dichloromethane B: Acetonitrile
Gradient: Time: 0 10 18 20
%B: 70 55 70 70
Flow Rate: 1.5mL/min
Detector: ELSD
Column: Alltech® Alltima C18, 3µm, 150 x 4.6mm (Part No. 81387)
Mobile Phase: A: Dichloromethane B: Acetonitrile
Gradient: Time: 0 10 18 20
%B: 70 55 70 70
Flow Rate: 1.5mL/min
Detector: ELSD
1. LLL 2. LLO 3. LLP 4. OOL 5. POL 6. PPL 7. OOO 8. OOP 9. PPO 10. OOS
1. Trilaurin (LaLaLa) 2. Trilinolein (LLL) 3. 1,2-Dilinoleyl-3-oleyl-rac-glycerol (LLO) 4. 1,2-Dilinoleyl-3-palmitin-rac-glycerol (LLP) 5. 1-Palmitin-2-olein-3-Linolein rac-glycerol (POL) 6. 1,2-Dipalmytyl-3-linolein-rac glycerol (PPL)
Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm
(Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichlor omethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
9522
9517
Column: Alltech® Alltima C18, 5µm, 250 x 4.6mm (Part No. 88056)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 40
%B: 30 50
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Prevail™ Silica, 3µm, 150 x 3.0mm
(Part No. 99341)
Mobile Phase: A: Chloroform
B: Methanol
C: Triethylamine buffer pH3
Gradient: Time: 0 16 17
%B: 12 60 12
%C: 0.5 12 0.5
Flow Rate: 0.5mL/min
Detector: ELSD
1. LLL 2. LLO 3. LLP 4. OOL 5. POL 6. PPL 7. OOO 8. OOP 9. PPO 10. OOS
10581
Column: HP C18 Hi-Load, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 1.0mL/min
Detector: ELSD
1. LGG 2. LGL 3. LLL 4. OGL 5. PGL 6. OLL 7. OGO + PLL 8. OOO
10047
Evening Primrose Oil
10566
1. C11 2. LLLn 3. LLL 4. LLO 5. PLL 6. LOO 7. PLO
8. PLP 9. OOO 10. SLO 11. POO 12. POP 13. SOO/PPP 14 C17
1. PA 2. PE 3. PI 4. LPE 5. PC 6. LPC
10562
Lecithin, 0.5mg/mL
Sesame Seed Oil
Walnut Oil
Sesame Seed Oil
Corn Oil
®
® ®
Alltech ® Alltima®Alltech
®
1
34
5
6
7
8
9
1011
12
1415 16
17
13
2
0 5 10 15 20 25 30 Min.
0 5 10 15 20 25 30 Min.
1
3
4567
8 910
11
2
1
2
3
4 5
6
7
8
9
10
1112
13
0 5 10 15 20 25 30 35 Min.
0 5 10 15 20 25 30 Min.
0 5 10 15 20 25 30 35 Min.
6
1
23 45 7
8
9
11
14
131210
0 5 10 15 20 25 30 35 Min.
1
3
4
56
7
89
2
10573
1. C13 2. PLO 3. PLP 4. POO 5. PLS 6. POP 7. SOO/PPP
10563
1056510564
Column: Alltech® Alltima HP C18 Hi-Load, 3µm,
150 x 3.0mm (Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 Hi-Load, 3µm,
150 x 3.0mm (Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm
(Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm
(Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
8. SLS 9. POS 10. PPS 11. SOS 12. PSS 13. SOA 14. C19
1. CCLa 2. C11 3. CLaLa 4. CCM 5. LaLaLa 6. CLaM
7. LaLaM 8. LaLaO 9. LaMM 10. LaMO 11. LaMP/MMM 12. MMP/LaPP/MPO 13. C17
1. CCLa 2. C11 3. CLaLa 4. CCM 5. LaLaLa 6. CLaM 7. LaLaM 8. LaLaO 9. LaMM
10. LaMO 11. LaMP/MMM 12. MMO/LaPO 13. MMP/LaPP/MPO 14. PLP 15. POO 16. POP 17. C17
1. C11 2. LLL 3. LLO 4. PLL 5. LOO 6. LLS 7. PLO 8. OOO 9. SLO 10. POO 11. C17
10569
1. LLL 2. LLO 3. PLL 4. LOO 5. LLS 6. PLO 7. OOO 8. SLO 9. POO
10567
Column: Alltech® Alltima HP C18 Hi-Load, 3µm,
150 x 3.0mm (Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 Hi-Load, 3µm,
150 x 3.0mm (Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Refined Sunflower OilSunflower Oil
Palm Kernel Olein Fraction Butter
Coconut Oil Cocoa Butter
Oils / Lipids
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® ®
® ®
Oils / Lipids
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hplc applications
6
4
3
52
1
0 5 10 15 20 Min.
0 5 10 15 20 25 30 35 Min.
1
3 4
56
78
9
10
11
12
1415
16
13
2
0 5 10 15 20 25 30 Min.
134
5
6
7
8
9
10
11 1213
2
0 5 10 15 20 25 30 35 Min.
1
3
4
5
6
7
8
9
101112
14
15
1716
18
19
132
6
4
3
7
85
21
0 5 10 15 20 Min.
6
4
3
7 8
910
5
21
0 5 10 15 20 Min.
1057210571
1. C13 2. OLL 3. PLL 4. LOO 5. PLO 6. PLP 7. MOP 8. OOO
9. POO 10. PLS 11. POP 12. SOO/PPP 13. POS 14. PPS 15. SOS 16. C19
Column: Alltech® Alltima HP C18 Hi-Load, 3µm,
150 x 3.0mm (Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm (Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm
(Part No. 87610)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 25 26 27 28
%B: 30 51 70 70 30
Flow Rate: 0.72mL/min
Detector: ELSD
1. CCLa 2. C11 3. CLaLa 4. CMM 5. LaLaLa 6. LaLaM 7. LaLaO
8. LaMM 9. LaMO 10. LaMP/MMM 11. MMO/LaPO 12. MMP/LaPP/MPO 13. C17
Column: Alltech® Prevail™ Silica, 3µm, 150 x 3.0mm (Part No. 99341)
Mobile Phase: A: Chloroform B: Methanol; C: Triethylamine buffer pH3
Gradient: Time: 0 16 17
%B: 12 60 12
%C: 0.5 12 0.5
Flow Rate: 0.5mL/min
Detector: ELSD
10570
Column: Alltech® Prevail™ Silica, 3µm, 150 x 3.0mm (Part No. 99341)
Mobile Phase: A: Chloroform B: Methanol C: Triethylamine buffer pH3
Gradient: Time: 0 16 17
%B: 12 60 12
%C: 0.5 12 0.5
Flow Rate: 0.5mL/min
Detector: ELSD
1. C11 2. LLnLn 3. LLLn 4. LLL 5. OLnL 6. PLLn 7. LLO 8. OOLn 9. PLL 10. LOO
11. LLS 12. PLO 13. PLP 14. OOO 15. SLO 16. POO 17. POP 18. SOO/PPP 19. C17
1. PE 2. PI 3. PS 4. PC 5. SM1 6. SM2
Butter
10578
1. GLUCCER 2. LACCER 3. PE 4. PI 5. PS 6. PC 7. SM1 8. SM2
Column: Alltech® Prevail™ Silica, 3µm, 150 x 3.0mm (Part No. 99341)
Mobile Phase: A: Chloroform B: Methanol C: Triethylamine buffer pH3
Gradient: Time: 0 16 17
%B: 12 60 12
%C: 0.5 12 0.5
Flow Rate: 0.5mL/min
Detector: ELSD
10575
Polar Lipid Standards
Palm Oil Palm Kernel Stearin Fraction
Soybean Oil
1. LACCER 2. U 3. PA 4. PE 5. PI 6. PS 7. PC 8. SM1 9. SM2 10. LPC
10577
Gouda Cheese
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1
0 2.5 5 7.5 10 12.5 15 17.5 20 Min.
0
0.5
1.0
1.5
2.0
mV(x100)
HPLC Method for Melamine
1
0 1 2 3 4 5 Min.
-1
1
2
3
mAU
0
4
5
UHPLC Method for Malamine
0 2.5 7.5 10 15 17.55 12.5 20 Min.
0
0.5
1.0
1.5
2.0
mV (x100)
UHPLC
HPLC
Melamine - HPLC Method Transfer to UHPLC
HILIC Column
Flow
Rate
Time
(min) Conc.
Inj.
Range
Loading
Range
HPLC®
UHPLC®
Melamine
0 2.5 7.5 10 15 17.55 12.5 20 Min.
0
100
200
300
mV
Melamine - ELSD Method
HPLC Column: ® 86466)Detector: ® 5135834)Mobile Phase:
Flow Rate:
Detection:
Column Temp:
ELSD 3300:
Injection:
Egg Yolk Phospholipids
Column: Allsphere™ Silica column, 3µm,
100 x 4.6mm
Mobile Phase: A: Isopropanol: Hexane: Water (58:40:2)
B: Isopropanol: Hexane: Water(52:40:8)
Gradient: Time: 0 7 19 20
%B: 0 100 100 0
Flow Rate: 1.25mL/min
Detector: ELSD
1. Phosphatidylethanolamine
2. Phosphatidylcholine
1
2
10214
Oils / Lipids
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Alltech
UHPLC Column: ® )
Mobile Phase:
Flow Rate:
Detection:
Column Temp:
ELSD 3300:
Injection:
HPLC Column: ® )
Mobile Phase:
Flow Rate:
Detection:
Column Temp:
Injection:
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600
790.361
1492.0711220.139
826.410
1012.464
507.293
1208.6 51050.523
804.370655.384
474.329 982.501
827.416
23.221 1041.482579.315 1119.553
1312.674847.384429.090752.342 1511.078
T7
T24
T19
T13
T8
T20
T11
T18-20
T17
T4
T5T26
T9
T10
Inte
nsi
ty (
co
un
ts)
m/z
Proteins & Peptides/Reversed-Phase
TIC of Tryptic Digest of Green Fluorescent Protein (GFP): 150pmol Load on 1mm i.d.
Column: Vydac®, 250 x 1mm (Part No. 238EV51)Mobile Phase: A: Acetonitrile:0.05% Trifluoroacetic Acid (2:98) B: Acetonitrile:0.05% Trifluoroacetic Acid (90:10)Gradient: Time: 0 60 %B: 5 65Flow Rate: 50µL/minDetector: ms: ABI QSTARInjection Volume: 25µL GFP digest
Mass Spectrum of Tryptic Digest of GFP
PTH Amino Acids
Column: Vydac® Denali®, 5µm, 150 x 4.6mm (Part No. 238DE5415)Mobile Phase: A: Acetonitrile:25mM Potassium Phosphate, pH3.3 (20:80) B: Acetonitrile:25mM Potassium Phosphate, pH3.3 (80:20)
Flow Rate: 1.0mL/minDetector: UV at 254nm
1. His 2. Cys Acid 3. Arg 4. Asp 5. Ser 6. Gln 7. Thr 8. Gly 9. Cys
10. Ala 11. Tyr 12. Met 13. Pro 14. Trp 15. Phe 16. Ile 17. Leu
Gradient: Time: 0 15 %B: 0 75
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0 2 4 6 8 10 12 14 16 Min.
13
2
4
5
6
1
2
3
4
0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 Min.
10369
1. Methionine Enkephalin 2. Leucine Enkephalin 3. Angiotensin II 4. Neurotensin 5. Angiotensin I 6. Substance P
Column: Alltech® Alltim HP C18, 190Å, 5µm, 250 x 4.6mm (Part No. 87679)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.08% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 20 %B: 20 40Flow Rate: 1.0mL/minDetector: UV at 220nm
Peptide Mix on 190Å
1. Monophosphopeptide (FQpSEEQQTEDELQDK)
2. [Glu1]-Fibrinopeptide B (EGVNDNEEGFFSAR)
3. Tetraphosphopeptide (RELEELNVPGEIVEpSLpSpSpSEESITR)
4. Contaminant(s)
Phosphopeptides and Fibrinopeptide
Column: Vydac® ProZap™ C18, 2.1 x 10mmHPLC System: HPG binary pumpMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in Acetonitrile:Water (80:20)Gradient: Time: 0.0 1.0 1.1 1.3 1.4 %B: 12 60 80 80 12Flow Rate: 0.8mL/minDetector: UV at 215nm
BSA Tryptic Digest10582
Amino Acids by HILIC
Column: Alltech® Alltima HP HILIC, 150 x 4.6mmMobile Phase: Acetonitrile:10mM Ammonium Acetate with 0.2% Acetic Acid (70:30)Flow Rate: 0.75mL/minDetector: ELSD
Proteins & Peptides/Reversed-Phase
0 1
1 34
5
67
2
2 3 4 5 6 7 8 9 10 11 Min.
10582
1. Glutamic Acid 2. Leucine 3. Serine 4. Glycine 6. Lysine 7. Arginine
Column: Vydac® 218MS C18, 5µm, 150 x 0.075mm (Part No. 218MS5.07515)Mobile Phase: A: Water with 0.1% Formic Acid:Acetonitrile (95:5) B: Water with 0.1% Formic Acid:Acetonitrile (20:80)Gradient: Time: 0 5 50 60 %B: 0 0 50 100Flow Rate: 0.25µL/min
1. (Arg-8)-Vasotocin2. (Lys-8)-Vasopressin3. Oxytocin4. Leucine Enkephalin5. Angiotensin 1
0 3
1 2
3
4
5
6 9 Min.
Column: Platinum™ 100Å 53 x 7mm Rocket™ Mobile Phase: A: 0.15% TFA in Water
B: 0.13% TFA in 95% Acetonitrile:5% WaterGradient: Time: 0 10
%B 20 50Flowrate: 3.0mL/minDetector: Alltech ELSD
8874
Column: Vydac® 208TP54, C8, 5µm, 4.6 x 250mm
Flow Rate: 1.0mL/min
Gradient: 0–30% ACN with 0.1% aqueous TFA over 60min
Separation of a Tryptic Digest on 208TP54
Reversed-Phase Column
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a®
® 3µm,EPS C18,
1
2
3
4
0 1 2 3 4 5 Min.
4 0 600 800 1000 1200 1400 1600
m/z
0
0.4
0.8
1.2
1.6
2.0
2.4
941.0
815.9874.0
765.1
1019.2
720.2
1112.1
447.0 1223.0
680.4 1358.4483.0
644.7
719.0417 0 541 9 945.8740.5 1120.26 4 0 996.2856 7 1528.3
707 5 888.669.8 1232.4
+19 +1 +17 +16 +15 +14 + 3 +12 +11 +10 +9
Peak 1 - Bovine Cytochrome C
907.1
879.2
852 9
935.4
881.1 1037.1
966.6
1000.0807.2
828.5 968 51116.4447.0
1161.2764.1
425.11266.8893.4
1319.3784.9744.1
1209.4416.9 871.5 948.1765.91038.3
1451.1505.0955.7726.1 799 2482.9 912.0 1060.1 1160 3
1381.9978.6689.3593 2467
1323.41052.5 1131.7691.8 930.7 1527.6992.3 1353.0525.9
630.3 776.6 1 52.4590.8 1612.2418.71330.01231 8 1477.2
1363.2
+ + + 0 + + + 7 + + + + + 2 +4 + +3 +3 +3 +3 +35 +34 +3 +32 +31 +30 +29 +28 +27 +26 +25 +24 +23 +22 +21 +20 +19
Peak 3 - Bovine Carbon c Anhydrase II
400 600 800 1000 1200 1400 1600
m/z
0 2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
1. Bovine Cytochrome C, M =12222 (ave.) 2. Horse Apomyoglobin, M = 16941 (ave.) 3. Bovine Carbonic Anhydrase II, M = 28998 (ave.) 4. Chicken Ovalbumin, M = 44455 (ave.)
Column: Vydac® ProZap™ C18, 2.1 x 10mm
Mobile Phase: A: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (5:95)
B: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (80:20)Gradient: Time: 0 4.0 4.5 4.7
%B: 20 90 90 20
Flow Rate: 0.4mL/min
Detector: AB/MDS Sciex Q TRAP® MS. EMS scan 400-1700 amu;
scan rate = 1000 amu/s; ion spray voltage = 5000 V; LIT fill time = 50 ms;
Q3 entry barrier = 8V; declustering potential = 20; collision energy = 10 eV;
curtain gas = 50; ion source gas 1 = 45; ion source gas 2 = 70;
temperature (TEM) = 350°C.
50 ng each
400 600 800 1000 1200 1400 1600
m/z
1
2
3
4
5
700 900 1100 1300 1500 1700
m/z
0
1
2
3
4
940 7
1012.2
961 8
1048.0997.7 1163.3
1140.31077 2 1348.31054.7
1482.81235.6 1521.6955.4
1387.9005.1919.3 1131.4615.9
1 34.11229.1967.5 1639.3
908.3 1117.81088.1 1396.01257.0
1200.3 1592 7945.8872.4
1282.0897.6 1351.11114.9 1515.4
1184.7 1471.9729.9 882 8 1643.9678.8621.0
1355.0 1 64.9 1527.01313.2
895.8 1480.2736.9 774.1 1248 2635.8 1655.9
1520.3 1637.8
694.5 1365.2753.0 843.5 1610.71502.3669.8
1569.6
1664.3
+35 +34 +33 +32 +31 +30 +29
Peak 4 - Chicken Ovalbumin
892.6848 1
942.2
808.0
997.2
1059.7
771.3
1304.01130.3737.9
1210.7
1412.6447.0 707.0 1322.61132.8895.2679.1 1000.9740.1425.0 805.0 1541.01216.9519.0 1694.9960.7541.0 715.6 1090.7 1245.9 1411.61011.8 1187.8 1578.6
+27 +26 +25 +24 +23 +22 +21 +20 +19 +18 +17 +16 +15 +14 +13 +12 +11
Peak 2 - Horse Apomyoglobin
LC-MS of Proteins
Proteins & Peptides/Reversed-Phase
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Injection Amount:
1
2
3
0 1 2 3 4 5 Min.
400 600 800 1000 1200 1400 1600
0.2
0.4
0.6
0.8
1.0
1451.61161 4
968.1
1453.1
1474 5827.6
1180.41158.0446.0 651.8 969 8608.8 1262.31040.1755.6 1465.1943.6 1340 1 150 .6817.9 1685.4
+7 +6 +5
Peak 1 -Human Insulin
m/z
Inte
nsi
ty (
x 1
06 c
ps)
400 600 800 1000 1200 1400 1600
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
1474.5
1579.71382.5
1302.1
1165.4
1229.11107.4
1054.4
1164.6
447.01006.3
1231.31108.6 1388.7
962.6483 0
504.9 664.21307.34 8.9
1201.1964.81136 3714 9
1374.0922.4524.9 1587.4885 6602.9
629.7 1493.81248.4 1396.8989.5 1127.3 1604.4
1032.9822.31528.7
+26 +25 +24 +23 +22 +21 +20 +19 +18 +17 +16 +15
Peak 3 - Human Growth Hormone
Inte
nsi
ty (
x 1
05 c
ps)
m/z
Inte
nsi
ty (
x 1
05 c
ps)
1100 1200 1300 1400 1500 1600 1700
0.4
0.8
1.2
1.6
2.0
1473.4
1542.8
1478 81387.2
1408 81446.9
1620.61331 9 1415 81513.0
1360.0 1444.91305.0
1356 5 1547.81281.2 1393.91389.7
1475.71509.8
1329.2 1420.91621.81255.1
1399.61452.21335.1
1282.7 1552 81492 8 585 01256.1
1464.81403.51363.5
1529.41323.6
1582.6 1646.61240.2 1489.31423.91213.7
1317.51537.4
1265.11599.81266.71189.2 1657 51505 8
1319.31376.3 1576.4
1168.5 1270.71694.31642.91220.2
1130.41150 7 1567.8
1143.7108 .51016 5 1678 7
+59 +58 +57 +56 +55 +54 +53 +52 +51 +50 +49 +48 +47 +46 +45 +44 +43 +42 +41
Peak 2 - Human Serum Albumin
m/z
1. Human Insulin, M = 5803 (ave.) 2. Human Serum Albumin, M = 66520 (ave.) 3. Human Growth Hormone, M = 22123 (ave.)
LC-MS of Proteins
Column: Vydac.® ProZap™ C18, 2.1 x 10mmMobile Phase: A: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (5:95) B: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (80:20)Gradient: Time: 0 4.0 4.5 4.7 %B: 20 90 90 20 Flow Rate: 0.4mL/minDetector: AB/MDS Sciex Q TRAP® MS. EMS scan 400-1700 amu; scan rate = 1000 amu/s; ion spray voltage = 5000 V; LIT fill time = 50 ms; Q3 entry barrier = 8V; declustering potential = 20; collision energy = 10 eV; curtain gas = 50; ion source gas 1 = 45; ion source gas 2 = 70; temperature (TEM) = 350°C.Injection Amount: 8µg each, except 1.5µg for growth hormone
Proteins & Peptides/Reversed-Phase
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20 40 60Min.
VYDAC® MS C4 Column1
2
Competitor 1 C4 Column1
2
Competitor 2 C4 Column1
2
Competitor 3 C3 Column1 2
VYDAC® MS C4 Column
1
2
3
4
Competitor 1 C4 Column12
3
4
Competitor 2 C3 Column1
2
3
4
Competitor 3 C4 Column1
42
Competitor 4 C4 Column1 2
4
20 65 Min.30 40 50 60
20 40 60 80 Min.
VYDAC® MS C18
Column
Competitor 1 C18
Column
Competitor 2 C18
Column
Columns: Vydac® MS C4 (Part No. 214MS54), Competitor 1 C4, Competitor 2 C4, and Competitor 3 C3 (all 300Å, 250 x 4.6mm packed with 5µm except 4µm for Competitor 1)Mobile Phase: 50mM Tris, pH7.5:n-Propanol (71:29)Flow Rate: 0.5mL/minDetector: UV at 220nm
Human Growth Hormone (hGH) Assay on 300Å C4 Columns
Columns: Vydac® MS C4 (Part No. 214MS54), Competitor 1 C4, Competitor 2 C3, Competitor 3 C4, and Competitor 4 C4 (all 300Å, 250 x 4.6mm packed with 5µm except 4µm for Competitor 1)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient Vydac®, Comp. 1, & Comp. 2: Time: 0 20 25 45 %B: 20 60 80 80Gradient Comp. 3 & Comp. 4: Time: 0 20 25 45 50 80 %B: 20 60 80 80 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm
Transmembrane Protein p14 on 300Å C4 Columns 1. 18.8 kD Protein 2. Non-myristoylated p14 3. Myristoylated p14 4. Triton™ X (Surfactant)
1. Desamido hGH 2. hGH
Columns: Vydac® MS C18 (Part No. 238MS5415), Competitor 1 C18, and Competitor 2 C18 (all 300Å, 5µm, 150 x 4.6mm)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 5 80 90 100 %B: 4 4 40 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm
Tryptic Digest of Fetuin on 300Å C18 Columns
Proteins & Peptides/Reversed-Phase
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0
1
2
3
4
5
6
7
0 5 15 25 35 45 55 65 75 85 95 Min.
Inte
nsi
ty (
x 1
06 c
ps)
1. Chicken 2. Bovine 3. Ovine 4. Rabbit 5. Human 6. Porcine 7. Rat I 8. Rat II
Base Peak Chromatogram of
Tryptic Digest of BSA
Insulin Variants
1. ApoC-IIIa 2. ApoC-IIIb 3. ApoC-IIa 4. ApoC-IIIc 5. ApoC-I 6. ApoC-IIb 7. ApoA-Ia 8. ApoA-Ib 9. ApoA-IIc 10. ApoA-IIa 11. ApoA-IIb
Human Apolipoproteins
Column: Vydac® Everest® C18, 300Å, 5µm, 300µm x 150mm (Part No. 238EV5.315)Mobile Phase: A: 0.1% Formic Acid in Water B: 0.1% Formic Acid in Acetonitrile Gradient: Time: 0 5 75 85 95 %B: 4 4 40 90 90Flow Rate: 5µL/minColumn Temp: 22ºCPeptide Load: 0.1µL injection of 5.8µg/µL (580ng or 9pmol)MS Detector: AB/MDS Sciex Q-Trap.
Tryptic Digest of Bovine Serum Albumin Tryptic Digest of Fetuin (a Glycoprotein)
Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 218TP54)Mobile Phase: A: Acetonitrile:0.1% Trifluoroacetic Acid (25:75) B: Acetonitrile:0.1% Trifluoroacetic Acid (58:42)Gradient: Time: 0 33 %B: 0 100Flow Rate: 1.2mL/minDetector: UV at 215nm
Column: Vydac® C4, 5µm, 250 x 4.6mm (Part No. 214TP54)Mobile Phase: A: 0.1% Trifluoroacetic Acid B: IsopropanolGradient: Time: 0 355 %B: 10 38Detector: UV at 220nm
Column: Vydac® Everest® C18, 5µm, 150 x 4.6mm (Part No. 238EV5415) Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 5 80 90 100 %B: 4 4 40 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm
Column: Vydac® Everest® C18, 5µm, 150 x 4.6mm (Part No. 238EV5415)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 5 80 90 100 %B: 4 4 40 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm
Column: Vydac® C4, 5µm, 250 x 4.6mm (Part No. 214TP54)Mobile Phase: A: 0.1% Trifluoroacetic Acid B: AcetonitrileGradient: Time: 0 25 %B: 27 30Detector: UV at 210nm
Ribosomal Proteins
Proteins & Peptides/Reversed-Phase
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Proteins & Peptides/Reversed-Phase
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Peptides on 238MS C18 Column with Various Modifiers
In order to demonstrate the effects of mobile phase modifiers and modifier concentrations, a mixture of six peptides was separated on a monomeric C18 MS column under identical gradient conditions with a variety of ion-pair reagents. Significant differences in selectivity arise depending on the concentrations and identities of the ions present in the mobile phase. Perfluorinated Acidic modifiers even at very low levels (0.005% w/v) produce increased retention. For this particular mixture, the best spread of peaks was obtained with heptafluorobutyric Acid (HFBA) at 0.02% (w/v). However, this might be expected to differ for other peptides.
Column: Vydac® Monomeric C18, 5µm, 250 x 4.6mm (Part No. 238MS54)Gradient: 10 to 40% Acetonitrile over 30 min for all chromatograms with constant concentration of modifier as indicated (w/v) Flow Rate: 1.0mL/minDetector: UV at 220nmSample: Oxytocin, Bradykinin, Angiotensin II, Eledoisin- related Peptide, Neurotensin, and Angiotensin I
0 0.2 0.4 0.6 0.8 1.0 1.2 Min.
5
2
1
24
3
2 4 53
1
0 0.2 0.4 0.6 0.8 1.0 Min.
1
2
0 1 2 3 4 5 6 Min.
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 Min.
1
2
3
4
5
6
7
Column: Vydac® ProZap™ C18, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 4.0 4.5 4.7 %B: 23 75 75 23 Flow Rate: 0.2mL/minDetector: UV at 280nm
Column: Vydac® ProZap™ C18, 500Å, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid:Acetonitrile (20:80)Gradient: Time: 0 0.1 0.7 1.1 1.2 %B: 4 15 20 50 4Flow Rate: 0.8mL/minDetector: UV at 215nm
1. Albumin 2. IgG
Antibodies in Sheep Serum on ProZap
Column: Vydac® ProZap™ C18, 500Å, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid:n-Propanol (10:90)Gradient: Time: 0 6.0 6.5 7.0 %B: 5 75 75 5Flow Rate: 0.5mL/minDetector: UV at 280nm
1. GY (238 Da) 2. VYV (379 Da) 3. Met Enkephalin (YGGFM, 573 Da) 4. Leu Enkephalin (YGGFL, 555 Da) 5. Angiotensin II (DRVYIHPF, 1045 Da)
Peptides
Column: Vydac® ProZap™ C18, 500Å, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid:Acetonitrile (20:80)Gradient: Time: 0 0.1 0.7 1.1 1.2 %B: 4 15 20 50 4Flow Rate: 0.8mL/minDetector: UV at 215nm
1. Contaminant Peptides/Impurities 2. RGAGGLGLGK-amide (883 Da) 3. Ac-RGAGGLGLGK-amide (926 Da) 4. Ac-RGVGGLGLGK-amide (954 Da) 5. Ac-RGVVGLGLGK-amide (996 Da)
Synthetic Peptides-10 Injections
1. Ribonuclease (37ng) 2. Bovine Insulin (37ng) 3. Cytochrome C (37ng) 4. Lysozyme (4ng) 5. Bovine Serum Albumin (37ng) 6. Carbonic Anhydrase (37ng) 7. Ovalbumin (37ng)
Fast Protein Analysis
Proteins & Peptides/Reversed-Phase
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Column: Vydac® C18, C4 and Diphenyl, all 5µm, 150 x 4.6mm
(Part Nos. 218TP5415, 214TP5415, 219TP5415)
Mobile Phase: A: 0.1% Trifluoroacetic Acid
B: Acetonitrile
Gradient: Time: 0 30
B%: 10 90
Sample: 18-Residue Helical Peptide (1) and
Six-helix Template- assembled Synthetic Protein (2)
Flow Rate: 1.0mL/min
Detector: UV at 215nm
Purification of Trityl-On Oligonucleotides
Comparison of Polypeptide Separations on
C18, C4 and Diphenyl Reversed-Phase Columns
1. H 2. TH6
Column: Vydac® Diphenyl, 5µm, 250 x 4.6mm
(Part No. 219TP54)
Mobile Phase: A: 0.1% Trifluoroacetic Acid
B: n-Propanol:Acetonitrile:0.1% Trifluoroacetic Acid (50:50)
Gradient: Time: 0 45
%B: 25 70
Flow Rate: 1.5mL/min
Detector: UV at 215nm
Separation of Protein / Peptide Mixture on
Vydac® 219TP Diphenyl Column
1. Horse Cytochrome C Tryptic Digest Fragments 2. Whale Myoglobin CNBr Digest Fragments 3. Ribonuclease 4. Cytochrome 5. Lysozyme 6. BSA 7. Myoglobin 8. Ovalbumin
Column: Vydac® C4, 5µm, 250 x 10mm (Part No. 214TP510)
Mobile Phase: A: 0.1M Triethylammonium Acetate, pH7.0 B: Acetonitrile
Gradient: Time: 0 14
B%: 16 23
Flow Rate: 5.0mL/min
Detector: UV at 260nm
Analysis of a Lipid Peptide
Column: Vydac® Diphenyl, 5µm, 250 x 4.6mm
(Part No. 219TP54)
Mobile Phase: A: 0.1% Trifluoroacetic Acid
B: Acetonitrile
Gradient: Time: 0 67.5
B%: 0 70
Flow Rate: 1.0mL/min
Detector: UV at 210nm
Proteins & Peptides/Reversed-Phase
Columns: Vydac® 218TP54 and 238TP54, both C18, 5µm,
4.6 x 250mm
Flow Rate: 1.0mL/min
Gradient: 10–40% ACN with 0.1% TFA (w/v) over 30min
Detector: UV at 220nm
Synthetic Peptides 1. GY 2. VYV 3. Neurotensin fragment 1-8 (pELYENKPR) 4. Ac-RGGGGLGLGK-NH2 5. RGAGGLGLGK-NH2 6. Ac-RGAGGLGLGK-NH2 7. Ac-RGVGGLGLGK-NH2 8. Oxytocin (CYIQNCPLG-NH2) 9. Met Enkephalin (YGGFM) 10. Bradykinin (RPPGFSPFR) 11. Ac-RGVVGLGLGK-NH2 12. Neurotensin Fragment 8-13 (RRPYIL) 13. Angiotensin II (DRVYIHPF) 14. Leu Enkephalin (YGGFL) 15. Neurotensin (pELYENKPRRPYIL) 16. Angiotensin I (DRVYIHPFHL)
Human Growth Hormone
1. di-Desamido HGH2. mono-Desamido HGH3. Human Growth Hormone (HGH)
Column: Vydac® C4, 5µm, 250 x 4.6mm (Part No. 214TP54)
Mobile Phase: Isopropanol:0.01M Tris buffer, pH7.5 (29:71)
Flow Rate: 1.0mL/min
Detector: UV at 220nm
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Min.
Analyses
Startingmaterial
Pure pool(fract 2 -16)
Impure pool(fract 1 &17-26)
Fraction Collectionfrom Run C
Fraction 1
Fractions 2-16
Fractions 17-26
A280
A) 26mg
B) 41.5mg
C) 60mg
0 25 7550 100 Min.
0 5 10 15 Min.
6x scale
6x scale
6x scale
40 60 807050 90 Min.
Column: Vydac® C4, 10-15µm, 10 x 250mm (Part No. 214TP101510)
A) Sample: 26mg partially purified porcine insulin in 30mL of 20%
Acetonitrile/0.1% Trifluoroacetic acid, pumped on at 8mL/min
Elution: 4mL/min
Gradient: 20-35% Acetonitrile in 0.1% Trifluoroacetic acid over 90 min
B) Sample: 41.5mg in 42mL of 20% Acetonitrile/0.1% Trifluoroacetic
acid, pumped on at 8mL/min
Elution: 4mL/min
Gradient: 20-27% Acetonitrile in 35 min, hold 27% for 10 min
followed by 27-28% in 20 min, 28%-35% in 10 min, and
35-95% in 5 min, all in 0.1% Trifluoroacetic acid.
C) Sample: 60mg in 60mL of 20% Acetonitrile/0.1% Trifluoroacetic
acid, pumped on at 8mL/min
Elution: 4mL/min
Gradient: 20-27% Acetonitrile in 35 min, hold 27% for 10 min
followed by 27-28.5% in 30 min, hold 28.5% for 20 min
Preparative Chromatography of Porcine Insulin Analyses of Starting Material and Pooled
Fractions from Preparative Column
Column: Vydac® C4, 5µm, 4.6 x 150mm
(Part No. 214TP5415)
Samples: Starting material and pooled fractions
of preparative run C, as indicated.
Detection: UV at 215nm
Elution: 1.5mL/min
Gradient: 30-33% Acetonitrile in 0.1%
Trifluoroacetic acid over 15 min
Proteins & Peptides/Reversed-Phase
0 1 2 Min.
1
2
3
4
5
6
7
Expedite™ Columns for Low Volume High Throughput LC
Column: ProZap™ C18, 1.5µm, 2.1 x 10mm Expedite™
Mobile Phase: A: 0.1% TFA in Water
B: 0.085% TFA in Acetonitrile
Flow Rate: 0.8mL/min
Detector: UV at 280nm
Injection Vol.: 5µL
1. Ribonuclease (37ng) 2. Bovine Insulin (37ng) 3. Cytochrome C (37ng) 4. Lysozyme (4ng) 5. Bovine Serum Albumin (37ng) 6. Carbonic Anhydrase (37ng) 7. Ovalbumin (37ng)
10736
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Column: Astec™ Chirobiotic® T™, 250 x 4.6mm (Part No. 12053)
Mobile Phase: Ethanol:Water (80:20)
Flow Rate: 1.0mL/min
Detector: UV at 210nm
Phenylalanine Enantiomers
Column: Alltech® Prevail™ C18, 5µm, 250 x 4.6mm (Part No. 99210)
Mobile Phase: A: 5mM Heptafluorobutyric Acid, 0.7% Trifluoroacetic Acid
B: Acetonitrile
Gradient: Time: 0 6 8 25
%B: 0 0 15 35
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Platinum™ EPS C18, 5µm, 150 x 4.6mm
(Part No. 32214)
Mobile Phase: 0.05M KH2PO4, pH3.2:Acetonitrile (65:35)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
10. Arg 11. Val 12. Met 13. Tyr 14. Ile 15. Leu 16. Phe 17. Trp
1. Gly 2. Ser/Asn 3. Asp 4. Gln 5. Ala/Thr 6. Glu 7. Cys/Lys 8. His 9. Pro
Amino Acids
Amino Acids without Derivatization
Thyroid Chemicals
8770
8282
1. DL-Tyrosine 2. 3-Iodo-Tyrosine 3. 3,5- Diiodo-D-Tyrosine
4. DL-Thyroxine 5. 3,3,5-Triiodo-L-Thyronine
Column: Standard Platinum™ 100Å C18, 3µm,53 x 7mm Rocket™ Column
Mobile Phase: A: 0.15% TFAB: Acetonitrile
Gradient: Time: 0 3 12 15%B 3 3 65 65
Flowrate: 2.5mL/minDetector: Alltech ELSD
Drift Tube Temp: 40°CNitrogen Flowrate: 1.75SLPM
1. Noradrenaline (norepinephrine)2. Adrenaline (epinephrine)3. Dopamine (3-Hydroxytyramine)4. 3,4-Dihydroxyphenylalanine (DOPA)5. Phenylalanine6. Homovanillic Acid (HVA)
0 1
1
2
2
3
3
4
4
5
5
6
6
7 Min.
8903
Catecholamines
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Amino Acids
0
1
2
3
4
56
7
8
9
10
11
13
14
1512
5 10 15 20 25 30 Min.
1. Aspartic Acid 2. Glutamic Acid 3. Serine 4. Histidine 5. Glycine 6. Threonine 7. Arginine 8. Alanine 9. Tyrosine 10. Methionine 11. Valine 12. Phenylalanine 13. Isoleucine 14. Leucine 15. Lysine
Column: OPA-HR, 5µm, 4.6 x 150mm (Part No. 28072)
Mobile Phase: A: Methanol B: Tetrahydrofuran
C: 20mM Sodium Acetate buffer, pH 5.5
Gradient: Time: 0 20
%A: 5 65
%B: 5 2
%C: 90 23
Flow Rate: 1.5mL/min
Detector: Fluorescence
OPA-Derivatized Amino Acids9234
Prevail™ Bonded Phases are Stable in 100% Aqueous Conditions
Catecholamines
1. Norepinephrine2. Epinephrine3. 5-Hydroxydopamine
Mobile Phase: 25mM KH2PO4, pH
Flow Rate: 1.0mL/min
Detector: UV at 270nm
9450
9452
After 24 hrs.
After 8 hrs.
After 20 min.
9451
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0 5 10 15 20 Min.
3
45
1
2
0 2 4 6 8 10 12 Min.
1
23
4
5
0 2 4 6 8 Min.
1
2
3
4
5
0 2 4 6 8 Min.
1
23
4
5
10347
1. Cytidine 2. Uridine 3. Xanthosine 4. Guanosine 5. Adenosine
Column: Alltech® Platinum™ EPS C18, 5µm, 250 x 4.6mm
(Part No. 32246)
Mobile Phase: A: 0.03M KH2PO4, pH3.2 B: Acetonitrile
Gradient: Time: 0 2 20
%B: 5 5 30
Flow Rate: 0.7mL/min
Detector: UV at 260nm
1. Cytidine 2. Uridine 3. Xanthosine 4. Guanosine 5. Adenosine
Column: Alltech® Alltima HP EPS C18, 5µm, 150 x 4.6mm
(Part No. 87715)
Mobile Phase: A: 0.03M Potassium Phosphate, pH3.2
B: Acetonitrile
Gradient: Time: 0 2 20
%B: 5 5 30
Flow Rate: 1.0mL/min
Detector: UV at 260nm
8245
Nucleic Acids
NucleosidesNucleosides
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: 0.025M KH2PO4, pH3.0:Acetonitrile (96:4)
Flow Rate: 1mL/min
Detector: UV at 254
1. Cytidine 2. Uridine 3. Xanthosine 4. Guanosine 5. Adenosine
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: 0.025M KH2PO4, pH3.0:Acetonitrile (98:2)
Flow Rate: 1.0mL/min
Detector: UV at 254
1. Cytosine 2. Uracil 3. Guanine 4. Adenine 5. Thymine
94119410
Nucleosides Nucleic Acid Bases
51 www.discoverysciences.com
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® ®
1
23
4
5
6
7
8
9
10
11
12
0 4 8 12 16 20 24 28 Min.
0 3
1
2
3
4
5 6
6 9 12 15 18 21 24 Min.
Column: Alltech® Platinum™ EPS C18, 3µm, 53 x 7mm Rocket™
(Part No. 50573)
Mobile Phase: 0.05M KH2PO4, pH3.0:Methanol (50:50)
Flow Rate: 2.3mL/min
Detector: UV at 254nm
1. Uracil 2. Benzamide 3. Benzyl Alcohol 4. Phenol 5. Methylbenzoate 6. Acetophenone 7. N,N-Diethyl-m-toluamide 8. Toluene 9. Benzophenone 10. Biphenyl
8982
Monoaromatics
7373
54409538
Column: Alltech® Alltima C18, 5µm, 150 x 4.6mm (Part No. 88052)
Mobile Phase: 0.05M KH2PO4, pH3.2:Methanol (60:40)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Column: Alltech® OA-1000, 9µm, 300 x 6.5mm (Part No. 9046)
Mobile Phase: 0.01N Sulfuric Acid
Flow Rate: 0.8ml/min
Column Temp: 65°C
Detector: RI
1. Glycerol 2. Methanol 3. Ethanol 4. Isopropanol 5. tert-Butanol 6. Propanol 7. sec-Butanol 8. Isobutanol 9. tert-Pentanol 10. Butanol 11. Isopentanol 12. Pentanol
1. 3-Amino-4-hydroxybenzoic Acid 2. 4-Hydroxybenzoic Acid 3. Benzoic Acid 4. 4-Phenylbutyric Acid 5. 5-Phenylbutyric Acid
Column: Alltech® Prevail™ Organic Acid, 5µm, 150 x 4.6mm
(Part No. 88640)
Mobile Phase: 0.025M KH2PO4, pH2.5:Acetonitrile (60:40)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
10314
1. 1,2-Phosphatidyl-ethanolamine 2. Phosphatidyl-DL-glycerol 3. Phosphatidylinositol 4. 1,2-Phosphatidyl-L-serine 5. Phosphatidylcholine 6. Sphingomyelin
1. DL-4-Hydroxy-3-methoxy-mandelic Acid 2. Homovanillic Acid 3. Vanillic Acid 4. Salicylic Acid 5. Benzoic Acid
Column: Alltech® Prevail™ Silica, 3µm, 150 x 4.6mm (Part No. 99275)
Mobile Phase: A: Isopropanol B: Hexane C: Water
Gradient: Time: 0 7 15
%B: 40 40 40
%C: 2 8 8
Flow Rate: 1.0mL/min
Detector: ELSD
Phospholipids
Miscellaneous
Benzyl Acids
Aromatic Acids
Alcohols
52www.discoverysciences.com
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0 1
1
2
3
4 5
6
7
2 3 4 5 Min
0 3
1
2
3
4
6 9 12 Min
0 2 4 6 8 10 12 14 16 18 20 Min.
2
13
45
6
7
10346
Column: Alltech® Platinum™ EPS C18, 1.5µm, 33 x 7mm Rocket™ (Part No. 50575)Mobile Phase: 0.025M KH2PO4, pH3.0:Acetonitrile (65:35)Flow Rate: 2.0mL/minDetector: UV at 254nm
Column: Alltech® Alltima HP EPS C18, 5µm, 150 x 4.6mm (Part No. 87715)Mobile Phase: Water:Methanol (60:40)Flow Rate: 1.0mL/minDetector: UV at 214nm
8671
Column: Alltech® Platinum™ C18, 5µm, 150 x 4.6mm (Part No. 32043)Mobile Phase: 0.025M K2HPO4, pH7.0:Methanol (25:75)Flow Rate: 1.0mL/minDetector: UV at 220nm
1. Methoxychlor 2. Dieldrin 3. Heptachlor 4. 4,4-DDT
8437
1. Simazine 2. Simetryn 3. Prometon 4. Atrazine 5. Ametryn 6. Prometryn 7. Terbutryn
8439
1. Diquat 2. Paraquat
Column: Alltech® Platinum™ Silica 100Å 3µm, 30mm x 4.6mm (Custom)Mobile Phase: 0.01M Tetramethylammonium Hydroxide and 0.15M Ammonium Sulfate in Water, pH3.0 with 5M Sulfuric AcidFlow Rate: 0.5mL/minDetector: UV at 310nm until 3 minutes UV at 260nm after 3 minutes
10355
Column: Alltech® Alltima HP CN, 5µm, 150 x 4.6mm (Part No. 87769)Mobile Phase: Water:Acetonitrile (60:40)Flow Rate: 1.0mL/minDetector: UV at 254nm
1. Fenuron 2. Monuron 3. Diuron 4. Linuron
1. Methyl Parathion 2. Dichlorvos 3. Ethoprophos 4. Azinphos
10739
1. Fenuron 2. Monuron 3. Fluometuron 4. Diuron 5. Methiocarb 6. Linuron 7. Neburon
Column: GraceSmart™ C18, 5µm, 150 x 4.6mm (Part No. 5138812)Mobile Phase: A: 0.025M KH2PO4, pH3.2 B: AcetonitrileGradient: Time: 0 20 %B: 30 60Flow Rate: 1.0mL/minDetector: UV at 240nm
Pesticides
Organophosphates Organochlorines
Triazine Herbicides Herbicides
Bipyridylium HerbicidesCarbamate and Urea Pesticides
53 www.discoverysciences.com
hplc applicationsenvironmental
®
®
n n
n n
0 2 4 6 8 10 12 14 Min.
1
2
0 1 2 3 4 5 6 7 8 9 Min.
1
2
34
56
7 89
10740
1. Methoxychlor 2. 4,4-DDT
Column: GraceSmart™ C18, 5µm, 150 x 4.6mm
(Part No. 5138812)
Mobile Phase: Methanol:0.025M K2HPO4, pH7.0 (75:25)
Flow Rate: 1.5mL/min
Detector: UV at 220nm
Organochlorine Pesticides
10580
Column: Vydac® Denali® 238DE C18, 3µm, 100 x 4.6mm
(Part No. 238DE3410)
Mobile Phase: A: 25mM Potassium Phosphate, pH6.4
B: Acetonitrile
Gradient: Time: 0 0.5 7.5 8.0
%B: 40 40 63 63
Flow Rate: 1.5mL/min
Detector: UV at 220nm
1. Simazine 2. Atraton 3. Simetryn 4. Atrazine 5. Prometon 6. Ametryn 7. Propazine 8. Prometryn 9. Terbutryn
Triazine Pesticides
Column: Vydac® Denali®, 3µm, 100 x 4mm (Part No. 238DE3410)
Mobile Phase: A: 0.5% Phosphoric Acid Buffer, pH1.8; B: Acetonitrile
Gradient: Time: 0 5
%B: 50 90
Flow Rate: 1.0mL/min
Detector: UV at 225nm
Separation of Chlorophenoxy Acid Herbicide Mixtures10713
Benzenes
1. Thiourea 2. Toluene 3. Propylbenzene 4. Butylbenzene
Column: Grace® VisionHT C18, 1.5µm, 50 x 2.0mm
(Part No. 5139605)
Mobile Phase: 20mM Potassium Phosphate, pH2.5:Acetonitrile (45:55)
Flow Rate: 0.8mL/min
Detector: UV at 254nm
Benzenes
1. Picloram 2. Chloramben 3. Dicamba 4. Bentazon 5. 2,4-D 6. Dichlorprop 7. 2,4,5-TP 8. Acifluorphen
0 0.5 1 1.5 2 Min.
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enviro
nmental
hplc applications
®
1
2
3
4
5 Min.
12 3
4
5
6
78
910
11
12
13
1415
16
0 1 2 3 4 5 6 Min.
Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)
Mobile Phase: A: Water B: Acetonitrile
Gradient: Time: 0 4 15
%B: 72 72 100
Flow Rate: 1.5mL/min
Detector: UV at 254
1. Benzene 2. Naphthalene 3. Acenaphthylene 4. Fluorene 5. Acenaphthene 6. Phenanthrene 7. Anthracene 8. Fluoranthene 9. Pyrene
10. Benz[a]anthracene 11. Chrysene 12. Benzo[b]fluoranthene 13. Benzo[k]fluoranthene 14. Benzo[a]pyrene 15. Dibenz[a,h]anthracene 16. Benzo[g,h,i]perylene 17. Indeno[1,2,3,c,d]pyrene
Column: Alltech® Alltima HP C18 HiLoad, 3µm, 53 x 7mm
(Part No. 87690)
Mobile Phase: A: Water B: Acetonitrile
Gradient: Time: 0 0.1 5
%B: 40 40 100
Flow Rate: 5mL/min
Detector: UV at 230nm
9400
1. Naphthalene 2. Acenapthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene
7. Fluoranthene 8. Pyrene 9. Benzo[a]anthracene 10. Chrysene 11. Benzo[b]fluoranthene 12. Benzo[k]fluoranthene
10757
PAHsPolyaromatic Hydrocarbons
Fast Polyaromatic Hydrocarbons
1. Naphthalene 2. Acenaphthylene 3. 1-Methylnaphthalene 4. 2-Methylnaphthalene 5. Acenaphthene 6. Fluorene 7. Phenanthrene 8. Anthracene
9. Fluoranthene 10. Pyrene 11. Benzo[c]phenanthrene 12. Cyclopenta[c,d]pyrene 13. Benz[a]anthracene 14. Chrysene 15. Benzo[b]naptho[2,1-d]thiopen 16. 7,12-Dimethylbenz[a]anthracene
Beyond the EPA 16 Priority-Pollutant PAHs
Column: Vydac® C18, 5µm, 150 x 4.6mm (Part No. 201TP5415)
Mobile Phase: A: Water B: Acetonitrile
Gradient: Time: 0 30
%B: 50 100
Flow Rate: 1.0mL/min
13. Benzo[a]pyrene 14. Dibenzo[a,h]anthracene 15. Benzo[g,h,i]perylene 16. Indeno[1,2,3,c,d]pyrene
17. Benzo[e]pyrene 18. Benzo[b]fluoranthene 19. Benzo[k]fluoranthene 20. Benz[a]pyrene 21. Dibenz[a,h]anthracene 22. Benzo[g,h,i]perylene 23. Indeno[1,2,3,c,d]pyrene
Detector: UV at 254
55 www.discoverysciences.com
hplc applicationsenvironmental
®
PAHs
10358
Column: Alltech® Alltima HP CN, 5µm, 150 x 4.6mm
(Part No. 87769)
Mobile Phase: Hexane:Isopropanol (85:15)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Nitrobenzene 2. 2,6-Dinitrotoluene 3. 1,3-Dinitrobenzene 4. 4-Nitrophenol
Nitroaromatics
Nitroaromatics (Normal-Phase)
1. Naphthalene 2. Acenaphthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene
Column: Vydac® C18, 5µm, 4.6 x 150mm
(Part No. 201TP5415)
Flow Rate: 1.5mL/min
Mobile Phase: A: Water B: ACN
Gradient: Hold 50% B for 3min, then 50 to 100% B in 7min
Detector: UV at 254nm
Priority Pollutants PAHs
In Accordance with
EPA Methods 505, 550.1, 610, and 8310
9. Benz[a]anthracene 10. Chrysene 11. Benzo[b]fluoranthene 12. Benzo[k]fluoranthene 13. Benzo[a]pyrene 14. Dibenz[ah]anthracene 15. Benzo[ghi]perylene 16. Indeno[1,2,3-cd]pyrene
enviro
nmental
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®
0 5 10 15 20 25 30 35 40 Min.
12
34
5
6
7
89
10
11
12 13 14
0
1
3
4
5
67
8
9
10
111213
15
16
14
17
18
192
10 20 30 40 50 Min.
2
1
34
5
0 2 4 6 8 10 12 14 Min.
0 2 4 6 8 10 12 14 16 18 20 22 Min.
12
3
4
5
6
7
89
10 11 12
13
Column: Alltech® Platinum™ EPS C18, 5µm, 250 x 4.6mm
(Part No. 32246)
Mobile Phase: A: 0.03M Monosodium Phosphate, pH2.5
B: Acetonitrile:Methanol:Water (40:50:10)
Gradient: Time: 0 3 13 20 25 30 40 45 55
%B: 5 5 10 10 20 40 75 80 80
Flow Rate: 1.0mL/min
Detector: UV at 210nm
1. 4-Amino-5-hydroxy-2,7-naphthalene-disulfonic Acid 2. Hydroquinone 3. Benzylamine 4. 3'-Aminoacetophenone 5. Phenol 6. 4-Hydroxyphenylacetic Acid 7. 1-Naphthalene Sulfonic Acid 8. 2,4-Dihydroxybenzoic Acid 9. 1-Naphthalene Sulfonic Acid Degradant 10. 4-Hydroxy-3-methoxybenzoic Acid 11. Phenylacetic Acid 12. m-Cresol 13. 4-Hydroxycinnamic Acid 14. trans-Cinnamaldehyde 15. trans-Cinnamaldehyde Degradant 16. trans-Cinnamaldehyde Degradant 17. 2-Naphthalenethiol Degradant 18. 2-Naphthalenethiol 19. 2-Naphthalenethiol Degradant
Column: Alltech® Adsorbosphere™ UHS C18, 5µm, 150 x 4.6mm
(Part No. 288118)
Mobile Phase: Water:Methanol (50:50)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. HMX
2. RDX
3. 1,3,5-TNB
Column: GraceSmart™ C18, 5µm, 250 x 4.6mm
(Part No. 5138810)
Mobile Phase: A: 1% Glacial Acetic Acid in Water
B: 1% Glacial Acetic Acid in Methanol
Gradient: Time: 0 6 8 15
%B: 45 45 60 80
Flow Rate: 1.0mL/min
Detector: UV at 280nm
4. 1,3-DNB 5. Tetryl 6. Nitrobenzene (NB) 7. 2,4,6-TNT
10511
8447
10707
Phenols
1. Phenol
2. 4-Nitrophenol
3. 2-Chlorophenol
4. 2-Nitrophenol
5. 2,4-Dichlorophenol
PhenolsSeparation of EPA 604 Phenols Mixture
1. Phenol 2. 4-Nitrophenol 3. 2-Chlorophenol 4. 2-Nitrophenol 5. 2,4-Dinitrophenol 6. 2,4-Dimethylphenol 7. 4-Chloro-3-Methylphenol 8. 2,4-Dichlorophenol 9. 2-Methyl-4,6-Dinitrophenol 10. 2,4,6-Trichlorophenol 11. Pentachlorophenol
Column: Vydac® Denali®, 5µm, 150 x 4.6mm (Part No. 238DE5415)
Mobile Phase: A: 1% Acetic Acid in Water
B: 1% Acetic Acid in Acetonitrile
Gradient: Time: 0 7 8 13
%B: 35 63 95 95
Flow Rate: 1.0mL/min
Detector: UV at 280nm
Column: Alltech® Platinum™ C18, 5µm, 250 x 4.6mm
(Part No. 32064)
Mobile Phase: Water:Methanol (50:50)
Flow Rate: 0.7mL/min
Detector: UV at 254nm
1. HMX
2. RDX
3. TNB
4. DNB
5. Tetryl
6. NB
7. TNT 8. 4-A-DNT 9. 2,6DNT 10. 2,4DNT 11. 2-NT 12. 3-NT 13. 4-NT
8279
8. 4-Amino-2,6-Dinitrotoluene (4-AM-DNT) 9. 2-Amino-4,6-Dinitrotoluene (2-Am-DNT) 10. 2,6-DNT 11. 2,4-DNT 12. 2-NT 13. 4-NT 14. 3-NT
Water PollutantsSemipolar & Highly Hydrophilic Waste Water Contaminants
Explosive Analysis, EPA Method 8330
Explosives
Nitroaromatic and Nitroamine Explosives
environmental
57
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Water Pollutants
8
Fullerenes
Column: Vydac® C4, 5µm, 250 x 10mm (Part No. 201TP510)
Mobile Phase: Acetonitrile:Toluene (50:50)
Flow Rate: 5mL/min
Buckminsterfullerenes
10310 1. n-octyl Glucopyranoside (50ng on column) 2. n-decyl Glucopyranoside (50ng on column) 3. n-dodecyl Glucopyranoside (50ng on column)
Column: Alltech® Alltima C18, 5µm, 150 x 1mm (Part No. 88384)
Mobile Phase: Methanol:Water (90:10)
Flow Rate: 0.05mL/min
Detector: ELSD
Microbore Analysis of Glucopyranosides
Surfactants
0 1
1
2
2
3
3
4 5 6 7 8 Min.
Column: Alltech® Alltima C18, 5µm, 250 x 4.6mm
(Part No. 88056)
Mobile Phase: Water:Methanol (40:60)
Flow Rate: 1.0mL/min
Detector: ELSD
8636
Polymers
0 4 8 12 16 20 Min.
Dimethicone
0 5 10 15 20 25 30 Min.
7809
Tween 80
Column: Alltech® Adsorbosphere™ C8, 5µm, 250 x 4.6mm
(Part No. 287112)
Mobile Phase: A: Methanol B: Water
Gradient: Time: 0 45
%B: 60 20
Flow Rate: 0.6mL/min
Detector: ELSD
8442
Polyethylene Glycol (MW=600)
0 5 10 15 20 25 30 35 Min.
miscella
neous
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Column: Alltech® Alltima C8, 5µm, 150 x 4.6mm (Part No. 88072)
Mobile Phase: A: Acetonitrile B: Chloroform
Gradient: Time: 0 20
%B: 25 75
Flow Rate: 1.0mL/min
Detector: ELSD
®®
Column: Alltech® Alltima C18, 3µm, 20 x 2.1mm Expedite™
(Part No. 43803)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.08% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 2
%B: 5 100
Flow Rate: 1.2mL/min
Detector: UV at 254nm
Columns: Alltech® Alltima C18, 3µm, 20 x 4.6mm Expedite™
(Part No. 43851)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.08% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 2
%B: 5 100
Flow Rate: 2.0mL/min
Detector: UV at 254nm
9462
9467
1. Uracil2. Theophylline3. Acetophenone4. Propiophenone5. Butyrophenone6. Benzophenone7. Valerophenone8. Heptanophenone9. Octanophenone
1. Uracil2. Theophylline3. Acetophenone4. Propiophenone5. Butyrophenone6. Benzophenone7. Valerophenone8. Heptanophenone9. Octanophenone
Phenones/Fast Gradient Column Formats Phenone Homologs
Phenone Homologs
1
0 1 2 3 Min.
2 3
4 5
6
7
89
0 1 2 3 Min.
1
2
3
4 5
6
7
8
9
Column: Alltech® Prevail™ C18, 3µm, 20 x 2.1mm, Expedite™
(Part No. 43827)
Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water
B: 0.08% Trifluoroacetic Acid in Acetonitrile
Gradient: Time: 0 2 3
%B: 0 100 100
Flow Rate: 1.0mL/min
Detector: UV at 254nm
9460
1. Uracil 2. Theophylline 3. Acetophenone 4. Propiophenone 5. Butyrophenone 6. Benzophenone 7. Valerophenone 8. Heptanophenone 9. Octanophenone
Phenone Homologs
miscellaneous
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® ®
0 105 25 Min.2015
12
3
5
4
6
9
10
1213
11
78
0 105 25 Min.2015
1
2
3
5
6
7
8
9
10
11 12 1314
15
4
0 5 10 15 20 25 30 Min.
1
9
8
76
5
4
32
1011
0 105 25 Min.2015
1 15
14
13
12
11
10 9
7
8
6
3
4
5
2
0 105 25 Min.2015
1
2
34
5
6
78
910
Biofuels
10644
Column: Alltech® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 5 30 32 35
%B: 0 15 70 70 0
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 20 25 30 35
%B: 30 70 70 30 30
Flow Rate: 1.0mL/min
Detector: ELSD
10748
1. LLnLn 2. LLLn 3. LLL 4. OLLn 5. PLLn 6. LLO 7. PLL 8. LLO
9. LLS 10. PLO 11. PLP 12. OOO 13. SLO 14. POO 15. POP
10749
Column: Alltech® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 20 25 30 35
%B: 30 70 70 30 30
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 20 25 30 35
%B: 30 70 70 30 30
Flow Rate: 1.0mL/min
Detector: ELSD
Column: Alltech® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm (Part No. 87698)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 20 25 30 35
%B: 30 70 70 30 30
Flow Rate: 1.0mL/min
Detector: ELSD
10750
1. PLL 2. LOO 3. PLO 4. PLP 5. MOP 6. OOO 7. POO 8. POP 9. SOO 10. PPP 11. POS 12. PPS 13. SOS
10751
1. LLnLn 2. LLLn 3. OLnL 4. LLO 5. LOO 6. PLO 7. OOO 8. POO 9. POP 10. PPP
1. LLnLn 2. LLLn 3. LLL 4. OLLn 5. PLLn 6. LLO 7. PLL 8. LOO
9. LLS 10. PLO 11. PLP 12. OOO 13. SLO 14. POO 15. POP
1. Mono-acylglycerol (MAG) 2. MAG 3. MAG
4. Fatty Acid Methyl Ester (FAME) 5. MAG 6. FAME 7. FAME 8. MAG 9. FAME 10. Di-acylglycerol (DAG) 11. Tri-acylglycerol (TAG)
Biodiesel in Spec with EN14105 Method
Triacylglycerols in Soy Oil
Triacylglycerols in Canola Oil Triacylglycerols in Red Palm Oil
Triacylglycerols in Used Chinese Frying Oil
miscella
neous
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®
® ®
0 105 25 Min.2015
1
2
3
4
0 2 4 6 8 Min.
1
5
7
64
3
2
0 2 4 6 8 Min.
2
4
3
1
1. Hexabromo Cyclododecane 2. Tetrabromo Bisphenol A (Allyl Ether)
Column: Alltech® Alltima C18, 5µm, 250 x 4.6mm (Part No. 88056)
Mobile Phase: Water:Acetonitrile (5:95)
Flow Rate: 1.0mL/min
Detector: ELSD
10261
Fire Retardants
Brominated Flame Retardants
10752
Column: ® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: A: Acetonitrile B: Dichloromethane
Gradient: Time: 0 20 25 30 35
%B: 30 70 70 30 30
Flow Rate: 1.0mL/min
Detector: ELSD
1. PPO 2. PPP 3. POS 4. SSS
Triacylglycerols in Lard
Biofuels
8760
1. Nicotine 2. Quinine 3. Diphenhydramine 4. Nortriptyline 5. 2,4, Dichlorophenoxy Acetic Acid 6. 2,4, Dichlorophenoxy Propionic Acid 7. 2,4, Dichlorophenoxy Butyric Acid
Column: GraceSmart™ C18, 5µm, 150 x 4.6mm (Part No. 5138812)
Mobile Phase: 50mM Potassium Phosphate, pH3.0:Acetonitrile (50:50)
Detector: UV at 254nm
10695
1. Uracil 2. Amitriptyline 3. Phenylvaleric Acid 4. Toluene
Column: GraceSmart™ C18, 5µm, 150 x 4.6mm (Part No. 5138812)
Mobile Phase: 50mM Potassium Phosphate, pH3.0:Acetonitrile (50:50)
Detector: UV at 254nm
StandardsStandards Mix Acids, Bases and Neutrals
Column:
Mobile Phase: 50mM KH2PO4, pH3.0:CH3CN (40:60)
Flowrate: 1.0mL/min
Detector: UV at 254nm
1. Uracil
2. Pyridine
3. Phenol
4. N,N-Dimethylaniline
5. 4-Butylbenzoic Acid
6. Toluene
Acids, Bases, and Neutrals
1
2
3
4
6
5
0 1 2 3 4 5 Min.
1. Phenylephrine
2. Diphenhydramine
3. Amitriptyline
0 1
1
2
2
3
3
4 5 Min.
Column: VisionHT C18-B, 1.5µm, 2.0 x 50mm (Part No. 5141908)
Mobile Phase: 50mM Ammonium Formate pH 7:Methanol (20:80)
Column Temp: 40°C
Flow Rate: 0.2mL/min
Detector: UV at 210nm
miscellaneous
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® ®
®
Alltech
® Alltima C18, 3µm, 100 x 4.6mm®Alltech
®
Separate Highly Basic Components at
Neutral pH with VisionHT C18-B
Standards
Column:
Mobile Phase: Acetonitrile:Water (58:42)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Final Acid washed with 24,000 column volumes of Acetonitrile:Dilute Sulfuric Acid, pH 1.0 (50:50) at 60°C.
Final Base washed with 24,000 column volumes of Acetonitrile Ammonium Hydroxide, pH 10 (50:50) at 20°C.
Retention Times
1. Uracil 0.56 0.55 0.55
2. Phenol 0.82 0.81 0.81
3. N,N-Diethyl-m-Toluamide 1.08 1.08 1.08
4. Toluene 1.89 1.88 1.89
1
2 3 4
1
23 4
0 1
1
2
34
2 3 Min.
Initial
Final Base
Final Acid
10252
Basic Compound Stability at pH1 and pH10
1025010249
0 2 4 6 8 Min.
1 2
3
6
4
5
EPS C18
C18 HiLoad
C18
12
3
4
5
612
3
4
5
6
1. Uracil 2. Pyridine 3. Phenol 4. N,N-diethyl-m-toluamide 5. 4-Butyl Benzoic Acid 6. Toluene
Column:
Mobile Phase: 50mM KH2PO4, pH 3.0:CH3CN (40:60)
Column Temp: Ambient (22°C)
Flow Rate: 1.0mL/min
Injection Vol.: 5µL
Detector: UV at 254nm
Acid, Base, and Neutral Test Mix
0 1 2 3 4 5 Min.
1
2
3
4
5
1. Toluene2. Hydantoin3. Uracil4. Caffeine5. Allantoin
10461
Column:
(Part No. 86464)
Mobile Phase: Acetonitrile:0.10M Ammonium Formate,
pH 3.0 (95:5)
Flow Rate: 1.0mL/min
Detector: UV at 205nm
Mixed Analyte HILIC
miscella
neous
62www.discoverysciences.com
hplc applications
® Alltima HP C18, 5µm, 4.6 x 50mm®Alltech
® Alltima HP , 4.6 x 150mm®Alltech ® Alltima HP HILIC, 5µm, 4.6 x 150mm®Alltech
0 2
2
1
4 6 Min.
Column: Alltech® Alltima C18, 5µm, 150 x 4.6mm (Part No. 88052)
Mobile Phase: 0.025M K2HPO4, pH7.0:Tetrahydrofuran:
Methanol (50:25:25)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
8584
1. 2,2’-Dipyridyl
2. 8-Hydroxyquinoline
Chelators
Metal Chelators
10429
Column: Alltech® Alltima HP C18 HiLoad, 3µm, 100 x 4.6mm
(Part No. 87685)
Mobile Phase: Acetonitrile:Dichloromethane (60:40)
Flow Rate: 1.0mL/min
Detector: ELSD
Paraffin Analysis
0 2 4 6 8 10 12 14 16 18 20 22 Min.
Paraffin C20-C30
0 2 4 6 Min.
1
4
5
6
2+3
C18-HL
2
3
1
4
5
6
C18
1
2
5
4
6
C18-P
3
C18-B
1
3
2
4
5+6
VisionHT
1. Uracil 2. Pyridine 3. Phenol 4. Methyl Alanine 5. 4-Butyl Benzoic Acid 6. Toluene
Column: 2.0 x 50mm
Mobile Phase: 50mM KH2PO4, pH 7.0:CH3CN (60:40)
Flow Rate: 1.0mL/min
Standards
miscellaneous
63
hplc applications
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Paraffins
Reversed-Phase Comparison®
® ®
0 4 8 12 16 20 24 Min.
1
2
3
4
5
67
8
9
10
0 2 4 6 8 10 12 14 16 18 20 22 24 26 Min.
1
2
3
4 5
6 7
8
9 10 11
1
2
3
0 2 4 6 8 10 12 14 16 18 20 22 24 Min.
0 2
3
2
1
4
4
6 8 10 Min.
0 5
1
10 Min.
2
ic applications
Anions
10120
1. Fluoride 2. Chlorite 3. Bromate 4. Chloride 5. Nitrite 6. Bromide 7. Chlorate 8. Nitrate 9. Phosphate 10. Sulfate
1. Fluoride 2. Chlorite 3. Bromate 4. Chloride 5. Nitrite-N 6. Bromide 7. Chlorate 8. Dichloroacetate 9. Nitrate-N 10. Phosphate-P 11. Sulfate
10121
Anions in Reagent Grade Water
by EPA Method 300.1Anions in High Ionic Strength Water by
EPA Method 300.1
1. Chlorite 2. Bromate 3. Nitrite
10123
Trace Bromate with Post Column Derivatization
and UV Detection, EPA Method 317.0
1. Aspartame 2. Chloride 3. Nitrate 4. Sulfate
1. Fluoride 2. Chloride 3. Phosphate 4. Monofluorophosphate
8458 8464
1. Arsenite 2. Arsenate
9178
Diet Cola Toothpaste
Arsenite and Arsenate
Column: Alltech® Novosep™ A-2 Anion, 250 x 4mm (Part No. 51410)
Mobile Phase: 3.6mM Sodium Carbonate
Flow Rate: 0.8mL/min
Column Temp: 43°C
Detector: Suppressed Conductivity
Column: Alltech® Novosep™ A-2 Anion, 250 x 4mm (Part No. 51410)
Mobile Phase: 3.6mM Sodium Carbonate
Flow Rate: 0.8mL/min
Column Temp: 43°C
Detector: Suppressed Conductivity
Column: Alltech® Novosep™ A-2 Anion, 250 x 4mm (Part No. 51410)
Mobile Phase: 3.6mM Sodium Carbonate
Flow Rate: 0.8mL/min
Column Temp: 40°C
Reagent: o-Dianisidine
Detector: UV at 450nm
Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)
Mobile Phase: 4mM Phthalic Acid, pH4.2 with Lithium Hydroxide
Flow Rate: 1.2mL/min
Detector: Conductivity
Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)
Mobile Phase: 0.85mM Sodium Bicarbonate/0.9mM Sodium Carbonate
Flow Rate: 1.2mL/min
Detector: Suppressed Conductivity
Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)
Mobile Phase: 4mM p-Hydroxybenzoic Acid, pH8.5
with Lithium Hydroxide
Flow Rate: 1.0mL/min
Detector: Conductivity
64www.discoverysciences.com
anions
Anions
65 www.discoverysciences.com
ic applicationsanions
0 2
1
2
3
45
6
4 6 8 10 Min.
1
2
3
4
5
6
7
0 2 4 6 8 10 12 14 16 18 20 22 24 Min. 0 1 2 3 4 5 6 7 8 9 Min.
1
2
1. Fluoride 2. Chloride 3. Nitrite 4. Bromide 5. Nitrate 6. Phosphate 7. Sulfate
1. Fluoride 2. Chloride 3. Nitrite 4. Bromide 5. Nitrate 6. Sulfate
1. Chloride 2. Nitrate 3. Phosphate 4. Sulfate 5. Oxalate
13033
84328451
Anions
Anions with Phthalic Acid Mobile Phase Oxalate in Urine
13040
1. Iodate 2. Bromate
Bakery Additives
8610 1. Fluoride 7. Selenite
ungstate Molybdate
Selenite, Selenate, Arsenate,
Tungstate and Molybdate
Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)
Mobile Phase: 0.85mM Sodium Bicarbonate/0.9mM Sodium Carbonate
Flow Rate: 1.2mL/min
Detector: Suppressed Conductivity
Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)
Mobile Phase: 0.85mM Sodium Bicarbonate/0.9mM Sodium Carbonate
Flow Rate: 1.2mL/min
Detector: Suppressed Conductivity
Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)
Mobile Phase: 4mM Phthalic Acid, pH4.2 with Lithium Hydroxide
Flow Rate: 1.0mL/min
Column Temp: 35°C
Detector: Conductivity
Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)
Mobile Phase: 0.85mM NaHCO3/ 0.9mM NaHCO3
Flow Rate: 1.0mL/min
Detector: Suppressed Conductivity
Column: Alltech® Allsep™ Anion, 150 x 4.6mm (Part No. 51209)
Mobile Phase: 0.85mM Sodium Bicarbonate/0.9mM Sodium Carbonate
Flow Rate: 1.2mL/min
Detector: Suppressed Conductivity
0
2
1
3
4 56
78
910
11 12
10 20 30 40 Min.
2. Chloride 3. Nitrite 4. Bromide 5. Nitrate 6. Phosphate
8. Sulfate 9. Arsenate 10. Selenate 11. T 12.
Column: Alltech® Anion/R, 10µm, 250 x 4.1mm (Part No. 269029)
Mobile Phase: 4mM Sodium Hydroxide/0.1mM Sodium Benzoate
Flow Rate: 1.5mL/min
1. Borate 2. Silicate 3. Formate 4. Sulfide 5. Chloride 6. Cyanide
5697
Weak Acid Anions1
23
4
5
6
0 9 18 Min.
Column: Alltech® Sarasep™ AN1™, 9µm, 250 x 4.6mm
(Part No. 38140)
Mobile Phase: 0.75mM Sodium Bicarbonate, 2.2mM Sodium Carbonate
Flow Rate: 1.0mL/min
Detector: Suppressed Conductivity
Fluoride Chloride Nitrite Bromide
Nitrate SelenitePhosphate
Sulfate Selenate
Column: Alltech® ICSep™ AN1™, 9µm, 250 x 4.6mm
(Part No. 38140)
Mobile Phase: 1.8mM Sodium Bicarbonate, 1.8mM Sodium Carbonate
Flow Rate: 1.0mL/min
Detector: Suppressed Conductivity
FluorideChloride
Bromide Nitrate Phosphate Sulfite
Sulfate
8611 6316
Selenite, Selenate, and Other Anions Separation of Sulfite and Sulfate
1. Fluoride 2. Bicarbonate 3. Chloride 4. Nitrite 5. Bromide 6. Nitrate 7. Phosphate 8. Sulfate
Column: Alltech® Anion/R, 10µm, 250 x 4.1mm (Part No. 269029)
Mobile Phase: 5mM p-Hydroxybenzoic Acid, pH8.5
with Lithium Hydroixde
Flow Rate: 1.5mL/min
Detector: Conductivity
6159Anion Standards
13038
Column: Alltech® OA-1000, 9µm, 300 x 6.5mm (Part No. 9046)
Mobile Phase: 1.0% Phosphoric Acid
Flow Rate: 0.8mL/min
Column Temp: 35°C
Detector: Conductivity (Non-Suppressed)
Haloacetic Acids Anions
1
2
3
4
5
6
7
8
0 9 18 Min. 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.
1
2
1. Chloro-difluoro Acetic Acid 2. Difluoro Acetic Acid
0 4
1
23
4 5
6
7
8
9
8 12 16 Min.
1. 2. 3. 4. 5. 6. 7. 8. 9.
1
2
34
5 6
7
8
9
0 2 4 6 8 10 12 14 16 Min.
1. Water 2. 3. 4. Nitrite 5. 6. 7. 8. 9.
0 4 8 12 16 20 24 Min.
1
2
3 4
1. Nitrite 2. EDTA 3. Bromide 4. Nitrate
Column: Anion/S, 10µm, 4.6 x 100mm (Part No. 269012)
Mobile Phase: 1.5mM Sulfuric Acid
Flow Rate: 1.0mL/min
Detector: UV at 210nm
EDTA and Other Anions
8284
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anions
ic applications
67 www.discoverysciences.com
cations
ic applications
0 5
1
2
3
4
5
10 15 20 Min.
8462
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 2.5mM Oxalic Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Sodium 2. Ammonium 3. Potassium 4. Magnesium 5. Calcium
Trace Level Ammonium in Brine
1
2
3
4
5
6
78
0 10 20 30 40 Min.
0 4
1
23
4 5 6
7
8 12 16 Min.
13013
6292
Column: Alltech® Universal Cation HR, 3µm, 53 x 7.0mm
(Part No. 23109)
Mobile Phase: 3mM Methanesulfonic Acid
Flow Rate: 2.5mL/min
Detector: Conductivity
1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Magnesium 6. Calcium
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 5mM Hydrochloric Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Rubidium 6. Magnesium 7. Calcium 8. Barium
Rapid Analysis of Monovalent
and Divalent Cations
Rubidium and Barium
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 3mM Methanesulfonic Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Morpholine 6. Magnesium 7. Calcium
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 5mM Hydrochloric Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Monoethanolamine 2. Diethanolamine 3. Triethanolamine 4. Magnesium 5. Calcium
8632
6315
Morpholine and Group I & II Cations
Ethanolamines
Cations
1
2
3
45
0 1 2 3 4 5 Min.
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.
1
23
4
5
6
7
8
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 Min.
1
2
3
4
5 67
89
0 2 4 6 8 10 12 14 16 18 20 Min.
1
23
4
5
6
78
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 5mM Hydrochloric Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Potassium 2. Magnesium 3. Calcium
Column: Hamilton® PRP™-X200, 10µm, 150 x 4.1mm
(Part No. 79441)
Mobile Phase: 4mM Nitric Acid in 30% Methanol
Flow Rate: 2.0mL/min
Detector: Conductivity
1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Cesium
6303 5204
Cations in Coffee Extract Alkali Metals
13035
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 2mM Tartaric Acid/1mM Oxalic Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Nickel 6. Zinc 7. Cobalt 8. Magnesium 9. Calcium
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 3mM Methanesulfonic Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Cesium 6. Magnesium 7. Calcium 8. Strontium
Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm
(Part No. 27106)
Mobile Phase: 3mM Tartaric Acid/2mM Oxalic Acid
Flow Rate: 1.0mL/min
Detector: Conductivity
1. Copper 2. Sodium 3. Ammonium 4. Potassium 5. Zinc 6. Lead 7. Magnesium 8. Calcium
10318 13042
Metal Ions
Cesium and Strontium
Transition Metals
Cations
68www.discoverysciences.com
catio
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ic applications
1
2
18 Min.0 4 8 12 14 162 6 10
0 2 4 6 8 10 12 14 16 18 20 Min.
1
2 3
4
Before SPE Clean-up
12
3
4
After SPE Clean-up
SPE Extraction of Salicylic and Acetylsalicylic Acids from Synthetic Urine
Column: Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm
(Part No. 87698)
Mobile Phase: 5mM Sodium Acetate, pH5.2:Methanol (80:20)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
1. Salicylic Acid 2. Acetylsalicylic Acid
10642
1. Oxazepam 2. Clonazepam 3. Temazepam 4. Diazepam
Benzodiazepines from Human Plasma
10606
10608
Analgesics
Sedatives
Procedure using GracePure™ C18-Aq, 500mg:
Sample Treatment – Spike 500µL plasma with 10µL of a 0.1mg/mL standard solution.
Combine with 100µL of 0.1M sodium carbonate buffer. Vortex and centrifuge for
10 minutes.
Conditioning – Rinse device 2mL water.
Sample Application – Apply entire sample.
Wash – Wash with 2mL water and then 50µL methanol.
Elution – Elute with 600µL methanol.
Column: Alltima HP C18, 5µm, 250 x 4.6mm (Part No. 87680)
Mobile Phase: A: Water B: Acetonitrile
Gradient: Time: 0 30
%B: 30 80
Flow Rate: 1mL/min
Detector: UV at 254nm
Procedure using GracePure™ C18-Max, 100mg:
Sample Treatment – Spike 2mL synthetic urine with 100ppm salicylic acid and 100ppm
acetylsalicylic acid.
Conditioning – Rinse device with 3mL methanol followed by 3mL DI water.
Sample Application – Apply 2mL spike urine.
Wash – Wash with 2mL 50mM phosphate buffer monobasic, pH 2.
Elution – Elute with 2mL methanol:water (50:50).
69 www.discoverysciences.com
spe applicationsp
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®
®
0 1 2 3 4 5 6 7 8 9 10 Min.
12
3
4
5
6
78
After SPE
Concentration
Before SPE
Concentration
0 1
1
2
2
3 Min.
0 1
1
2 3 Min.
spe applications
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nutra
ceutic
als
1. Acetazolamide 2. Hydrochlorothiazide 3. Hydroflumethiazide 4. Clopamide 5. Trichloromethiazide 6. Indapamide 7. Bendroflumethiazide 8. Spironolactone
Diuretics from Urine
10289
10290
Diuretics
Column: Alltima™ C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)
Mobile Phase: Methanol:0.2% Phosphoric Acid (95:5)
Flow Rate: 4.0mL/min
Detector: VIS at 585nm
1. Pseudohypericin 2. Hypericin
9107
Hypericins from St. John’s Wort
Column: Alltima™ C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)
Mobile Phase: Acetonitrile:20mM K2HPO4, pH7.3 (65:35)
Flow Rate: 3mL/min
Detector: UV 340nm
Inj. Vol.: 5µL
1. Apigenin-7-Glucoside
9133
Antioxidants from Chamomile
Procedure Using GracePure™ C18-Max, 1000mg:
Sample Treatment – Pulverize 300mg St. John’s Wort powder into five, 3mL aliquots
of methanol:water (80:20). Combine and filter extracts to produce 15mL filtrate. Dilute
to 30mL with water.
Conditioning – Rinse device with 5mL methanol followed by 5mL deionized water.
Sample Application – Apply 2mL filtrate to the top of the SPE device and draw through
the C18 packed bed.
Wash* – Wash device with 3mL of deionized water.
Elution – Elute with 2mL methanol.*Repeat load and wash steps consecutively until filtrate is consumed.
Procedure Using GracePure™ C18-Max, 1000mg:
Sample Treatment – Pulverize 1g commercial chamomile tea grounds into
6mL dioxane:methanol (50:50). Filter extract and dilute to 20mL with water.
Conditioning – Rinse device with 5mL methanol followed by 5mL water.
Sample Application* – Apply 1mL filtrate.
Wash* – Wash with 1mL water.
Elution – Elute first with 2mL methanol:water (50:50). Elute second, fraction
and use for this analysis, with 2mL methanol.*Before elution step, repeat load and wash steps until filtrate is consumed.
St. John’s Wort
Procedure using GracePure™ C18-Max, 1000mg:
Sample Treatment – Spike synthetic urine with 8 diuretics to a concentration of
1.25µg/mL each.
Conditioning – Rinse device 5mL methanol, followed by 5mL water.
Sample Application – Apply 15mL spiked urine sample at 1mL/min.
Wash – Wash with 5mL water.
Elution – Elute with 2mL methanol. Evaporate solvent and reconstitute in 250µL
HPLC mobile phase.
Chamomile
Column: Alltima™ C18, 3µm, 100 x 4.6mm (Part No. 81382)
Mobile Phase: A: 25mM Ammonium Acetate, 0.1%Trifluoroacetic Acid
B: Acetonitrile, 0.1%Trifluoroacetic Acid
Gradient: Time: 0 10
%B: 20 90
Flow Rate: 1.0mL/min
Detector: ELSD
0 1
1
2
2
3
3
4
4
5
5
0 1
1
2 3 4 5 Min.
0 1
1
2
3
2 Min.
Column: Alltima C18, 3µm, 53 x 7mm Rocket™ (Part No. 50605)
Mobile Phase: Acetonitrile:Isopropyl Alcohol:0.2% Acetic Acid (17:23:60)
Flow Rate: 3.5mL/min
Detector: UV 220nm
Inj. Vol.: 5µL
1. Methysticin 2. Yangonin 3. Kavain 4. Dihydrokavain 5. Dihydromethysticin
9125
Sedatives from Kava Kava
Column: Alltima C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)
Mobile Phase: Methanol:0.2% Acetic Acid (65:35)
Flow Rate: 4.5mL/min
Detector: UV 270nm
Inj. Vol.: 5µL
1. Z-ligustilide
9132
Vasodilators from Dong Quai
Column: Alltima C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)
Mobile Phase: Acetonitrile:10mM K2HPO4, pH2.6 (20:80)
Flow Rate: 3.5mL/min
Detector: UV at 330nm
1. Echinicoside 2. Chlorogenic Acid 3. Caffeic Acid
9110
Phenolic Acids from Echinacea
Procedure Using GracePure™ C18-Max, 1000mg:
Sample Treatment – Pulverize 300mg echinacea powder into five, 3mL aliquots of
methanol. Combine and filter extracts to produce 15mL filtrate. Dilute to 30mL with
water.
Conditioning – Rinse device with 5mL methanol followed by 5mL deionized water.
Sample Application – Apply 2mL filtrate to the top of the SPE device and draw through
the C18 packed bed.
Wash* – Wash device with 2mL of deionized water.
Elution – Elute with 5mL of methanol:water (50:50).*Repeat load and wash steps consecutively until filtrate is consumed.
Procedure using GracePure™ C18-Max, 1000mg:
Sample Treatment – Pulverize 300mg commercial kava kava root powder into four, 2mL
aliquots of methanol:water (80:20). Combine and filter extracts to produce 8mL filtrate.
Dilute to 10mL with water.
Conditioning – Rinse device with 5mL methanol followed by 5mL water.
Sample Application* – Apply 2mL filtrate.
Wash* – Wash with 2mL water.
Elution – Elute with 3mL methanol.*Before elution step, repeat load and wash steps until filtrate is consumed.
Echinacea
Kava Kava
Dong Quai
Procedure using GracePure™ C18-Max, 1000mg:
Sample Treatment – Pulverize 300mg commercial kava kava root powder into four, 2mL
aliquots of methanol:water (80:20). Combine and filter extracts to produce 8mL filtrate.
Dilute to 10mL with water.
Conditioning – Rinse device with 5mL methanol followed by 5mL water.
Sample Application* – Apply 2mL filtrate.
Wash* – Wash with 2mL water.
Elution – Elute with 3mL methanol.*Before elution step, repeat load and wash steps until filtrate is consumed.
71 www.discoverysciences.com
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®
®
0 10 20 30 35 40 45 50 605 15 25 55 65 Min.
Before SPE Clean-up
1
2
6
34
5
After SPE Clean-up
Column: Heliflex® AT™ AquaWax-DA, 30m x 0.25mm x 0.25µm
Capillary GC Column (Part No. 14537)
Temp: 200°C (5 min hold) to 230°C (4 min hold) at 5°C/min
Carrier Gas: Helium at 0.75mL/min (25cm/sec)
Detector: FID at 250°C
1. Sorbic Acid 2. Benzoic Acid
2576
Preservatives from Fruit Punch
Additives
Procedure using GracePure™ Anion-X, 500mg:
Sample Treatment – Use 8mL fruit punch and adjust pH to 10 using potassium
hydroxide.
Conditioning – Rinse device 10mL water.
Sample Application – Apply 8mL pH adjusted fruit punch sample.
Wash – Wash with 20mL water.
Elution – Elute with 1mL 1.0N hydrochloric Acid followed by 1mL methanol.
1. Carbendazim 2. Thiabendazole 3. Triadimenol 4. Triadimefon 5. Flusilazole 6. Diniconazole
Fungicides from Red Wine
10637
10638
Carbohydrates
Procedure using GracePure™ C18-Max, 500mg:
Sample Treatment – Add 0.167mg/mL each of carbendazim, thiabendazolem,
triadimenol, triadimefon, flusilazole, diniconazole into
1mL water. Combine with 1mL Beaujolais red wine.
Conditioning – Rinse device with 3mL methanol followed by 3mL water.
Sample Application – Apply 2mL red wine mixture.
Wash – No wash.
Elution – Elute with 3mL methanol.
Column: Alltima HP C18 Amide, 5µm, 250 x 4.6mm
(Part No. 87734)
Mobile Phase: A: Water B: Acetonitrile
Gradient: Time: 0 15 50 65
%B: 15 45 45 15
Flow Rate: 1mL/min
Detector: UV at 254nm
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Melamine
0 2 4 6 10 128 14 Min.
0
40
80
120
160
mAU
Melamine from Biscuit
SPE Column Type: ™ 5138770)
Sample Pretreatment: weight biscuit 2g
melamine standard solution
Mobile Phase:
Column: ®
Detector:
Flow Rate:
Column Temperature:
Injection:
SPE Column Type: ™ 5138770)
Sample Pretreatment: weight pork 2g
melamine standard solution
SPE recovery*=103%
HPLC Mobile Phase:
Column: ®
Detector:
Flow Rate:
Column Temperature:
Injection:
SPE Steps:
Column Conditioning: 2 column volumes methanol, then 2 column volumes water
Sample Application:
Column Washing: 3 column volumes water, then 2 column volumes methanol
Eluent:
Melamine standard
Spiked pork before SPE
Spiked pork after SPE
SPE Steps:
Column Conditioning: 2 column volumes methanol, then 2 column volumes water
Sample Application:
Column Washing: 3 column volumes water, then 2 column volumes methanol
Eluent:
Melamine from Pork
0 2 4 6 10 128 14 Min.
0
40
80
120
160
mAU
SPE recovery*=80%
Melamine standard
Spiked biscuit before SPE
Spiked biscuit after SPE
0 2 4 6 10 128 14 Min.
0
50
100
150
200
mAU
Melamine from HamSPE Column Type: ™ 5138770)
Sample Pretreatment: weight ham 2g
melamine standard solution
SPE recovery*=92%
Mobile Phase:
HPLC Column:
Detector:
Flow Rate:
Column Temperature:
Injection:
SPE Steps:
Column Conditioning: 2 column volumes methanol, then 2 column volumes water
Sample Application:
Column Washing: 3 column volumes water, then 2 column volumes methanol
Eluent:
Melamine standard
Spiked ham before SPE
Spiked ham after SPE
73 www.discoverysciences.com
spe applicationsfo
od
& b
eve
rag
e
®
1
2
3
0 1 2 3 4 5 6 7 8 9 Min.
1. Chloride 2. Ribonuclease A 3. Cytochrome C
10688
Prevail™ SPE Protein Desalting, Cytochrome C and Ribonuclease A Mix in 10mM Sodium Chloride
Protein Extraction of Ribonuclease and Myoglobin
1. Ribonuclease 2. Myoglobin
Proteins & Peptides/Reversed Phase
Procedure using Vydac® SPE:
A 3mL SPE cartridge was conditioned with 1mL of Acetonitrile followed by 1mL of
5% Acetonitrile/0.1% Trifluoroacetic Acid. Ribonuclease and myglobin (100mg each)
were then loaded in 30% Acetonitrile/0.1% Trifluoroacetic Acid . The cartridge was
washed with 1mL of 5% Acetonitrile/0.1% Trifluoroacetic Acid to remove weakly bound
compounds, then 1mL of 30% Acetonitrile/0.1% Trifluoroacetic Acid followed by 1mL of
60% Acetonitrile/0.1% Trifluoroacetic Acid. HPLC analysis of the 5% Acetonitrile wash
(A) revealed only a small amount of ribonuclease. Most of the ribonuclease eluted in the
30% Acetonitrile wash (B). The myoglobin eluted almost entirely in the 60% Acetonitrile
wash (C).
Procedure using Prevail™ C18, 500mg:
Sample Treatment – Cytochrome-C 1mg/mL, Ribonuclease-a 1mg/mL, Blank = water,
Salt = 10mM NaCl.
Conditioning – 1 tube volume of methanol (do not dry). 2 tube volumes of 0.025%
ammonium hydroxide. Dry tube via vacuum.
Sample Application – Pipette 1mL of sample (either proteins in water blank or proteins
in salt solution) onto top of sorbent bed in SPE tube. Run vacuum @ approximately
-10 kPa, loading sample in a tight band at top of sorbent column, continue to apply
vacuum (@ -10 kPa) until tube is dry. Larger (2mL +) volumes can be de-salted and
concentrated. Liquid collected will contain the NaCl
Elution – Pipette 500µL 0.4% TFA and 500µL Acetonitrile containing 0.4% TFA onto top
of sorbent bed in spe tube. Run vacuum @ approximately -10 kPa, eluting sample from
sorbent column, continue to apply vacuum until tube is dry.
Column: ProSphere™ 300 C4-300, 5µm, 150 x 4.6mm
(Part No. 33989)
Mobile Phase: A: 0.2% Trifluoroacetic Acid B: Acetonitrile
Gradient: Time: 0 15
%B: 25 70
Flow Rate: 1mL/min
Detector: ELSD
Column: Vydac® C4, 5µm, 50 x 4.6mm (Part No. 214TP5405)
Mobile Phase: A: 0.1% Trifluoroacetic Acid B: Acetonitrile
Gradient: Time: 0 10
%B: 15 70
spe applicationslife
scie
nce
s
74www.discoverysciences.com
Before SPE Clean-up
After SPE Clean-up
0 10 20 305 15 25 35 Min.
123
45
6
7 8
9101211
13
15
14
16
Before SPE Clean-up
After SPE Clean-up
0 10 20 305 15 25 35 Min.
12
3
4
5 6
78
109 1112
131415+16
After SPE Clean-up
0 5 10 15 20 25 30 35 40 Min.
Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)
Temp: 160°C (0 min hold) at 10°C/min, then to 275°C at 5°C/min
Carrier Gas: Helium at 0.8mL/min (26cm/sec)
Detector: ECD at 310°C
Chlorinated Pesticides in Vegetables2780
2789
Chlorinated Pesticides in Soil
2779
2788
1. Aroclor 1254
PCBs in Transformer Oil
Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)
Temp: 160°C (0 min hold) at 10°C/min, then to 275°C at 5°C/min
Carrier Gas: Helium at 0.8mL/min (26cm/sec)
Detector: ECD at 310°C
Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)
Temp: 150°C (2 min hold) to 300°C (8 min hold) at 5°C/min
Carrier Gas: Helium at 0.75mL/min (25cm/sec)
Detector: ECD at 310°C
2784
Pesticides
Procedure using GracePure™ Florisil®, 1000mg:
Sample Treatment – Dissolve 0.25grams of transformer oil spiked with Aroclor 1254 (at
a concentration of 50mg/kg) in 20mL n-hexane.
Conditioning – 2 x 5mL n-hexane, making sure the cartridge does not dry.
Sample Application – Attach 25mL reservoir (Part No. 210425) with a syringe adapter
(Part No. 210705) and run sample through conditioned Florisil® tube, aspirating all the
solution from the tube. Evaporate the volume down to 4mL and analyzed by GC/ECD.
Wash – No wash.
Elution – No elution, collect sample load.
Procedure using GracePure™ C18-Max, 500mg:
Sample Treatment – Homogenize 5 grams green bell pepper with 25mL methanol, spike
with 0.5µg each of 16 chlorinated pesticides. Filter with 75mL filter column (Part No.
210775). Dilute to 200mL with DI water and shake.
Conditioning – 2 x 5mL methanol, then 5mL DI water.
Sample Application – Attach 75mL reservoir (Part No. 210575) with a syringe adapter
(Part No. 210705) and apply entire sample.
Wash – No wash, apply vacuum for 5 min.
Elution – Elute chlorinated pesticides with 2 x 1mL n-hexane/CH2Cl2 (70:30),
evaporated this to dryness and reconstituted with 1mL n-hexane.
Procedure using GracePure™ C18-Max, 500mg:
Sample Treatment – Weighed out 5 grams dry soil, spike with 1µg each of 16
chlorinated pesticides, add added 25mL methanol and homogenize. Filter with 75mL
filter column (Part No. 210775). Dilute to 200mL with DI water and shake.
Conditioning – 2 x 5mL methanol, then 5mL DI water.
Sample Application – Attach 75mL reservoir (Part No. 210575) with a syringe adapter
(Part No. 210705) and apply entire sample.
Wash – No wash.
Elution – Elute chlorinated pesticides with 2 x 1mL n-hexane/CH2Cl2 (70:30), evaporate
to dryness and reconstitute with 1mL iso octane.
1. -BHC 2. -BHC 3. -BHC 4. -BHC
5. Heptachlor 6. Aldrin 7. Heptachlor Epoxide 8. Endosulfan I 9. p,p’-DDE
10. Dieldrin 11. Endrin 12. Endosulfan II 13. p,p’-DDD 14. Endrin Aldehyde 15. p,p’-DDT 16. Endosulfan Sulfate
1. -BHC 2. -BHC 3. -BHC 4. -BHC 5. Heptachlor
6. Aldrin 7. Heptachlor Epoxide 8. Endosulfan I 9. p,p’-DDE 10. Dieldrin
11. Endrin 12. Endosulfan II 13. p,p’-DDD 14. Endrin Aldehyde 15. p,p’-DDT 16. Endosulfan Sulfate
75 www.discoverysciences.com
spe applicationsenvironmental
Before SPE Clean-up
After SPE Clean-up
0 10 20 305 3515 25 40 Min.
0 9 18 Min.
12
3
45
6
7
89
10
11
12
13
14
15
16 17
18
0 5 10 15 20 25 Min.
Before SPE
Concentration
6
5
4
321
After SPE
Concentration
1. Aroclor 1242
PCBs in Salmon
2781
2787
Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)
Temp: 150°C (2 min hold) to 300°C (2 min hold) at 5°C/min
Carrier Gas: Helium at 0.75mL/min (25cm/sec)
Detector: ECD at 310°C
Pesticides
Procedure using GracePure™ C18-Fast, 1000mg:
Sample Treatment – Weigh out 2.5 grams salmon, spike with 2.5µg Aroclor 1242, add
25mL acetonitrile, homogenize for 30 seconds. Filter with 75mL filter tube (Part No.
210775). Rinse salmon again with 5mL acetonitrile. Bring up to 100mL with deionized
water.
Conditioning – Rinse with 6mL hexanes, 6mL acetone, 2 x 6mL methanol, 2 x 6mL
water.
Sample Application – Attach 75mL reservoir, apply entire sample.
Wash – 2 x 5mL water. Dry columns for 10 minutes at full vacuum.
Elution – Elute PCBs with 2 x 3mL 3% toluene/97% n-hexane. Evaporate
solvent and reconstitute in 1mL n-hexane.
Chlorinated Pesticides from Water
Column: AT™ Pesticide 20m x 0.53mm x 0.60µm Capillary GC
Column (Part No. 16846)
Temp: 210°C
Carrier Gas: Helium, 35cm/sec
Detector: ECD
Procedure using GracePure™ C18-Fast, 1000mg:
Conditioning – Rinse device with 5mL of methanol followed by 5mL deionized water.
Sample Application – Pass 100mL–500mL (containing 1% methanol) of water sample
through the device at 20mL/minute.
Wash – Wash device with 10mL of deionized water then 10mL of methanol:deionized
water (20:80). Remove excess by passing air through the device for two minutes.
Elution – Elute with 2mL of hexane:ethyl acetate (70:30). Pass extract through 2g–3g
sodium sulfate to remove residual water.
1668
Recovery (as % Standard) 1. -BHC (104%) 2. Lindane (109%) 3. -BHC (106%) 4. Heptachlor (92%) 5. -BHC (110%) 6. Aldrin (98%) 7. Heptachlor Epoxide (107%) 8. -Endosulfan (105%) 9. p,p'-DDE (79%) 10. Dieldrin (102%) 11. Endrin (104%) 12. p,p'-DDD (92%) 13. -Endosulfan (103%) 14. p,p'-DDT 15. Endrin Aldehyde (102%) 16. Endosulfan Sulfate (103%) 17. Methoxychlor (99%) 18. Endrin Ketone (104%)
Carbamate Pesticides from Water
Procedure using GracePure™ C18-Fast, 500mg:
Sample Treatment – Spike 500mL tap water with 125µL carbamate solution for final
concentration of 25ppb.
Conditioning – Rinse with 3mL acetonitrile:water (80:20) followed by 3mL water. Dry
with vacuum.
Sample Application – Apply 500µL sample.
Wash – 2 x 3mL water.
Elution – Elute with 4 x 1mL acetonitrile:water (80:20)
10599
10600
1. Aldicarb sulfoxide 2. Methomyl 3. 3-Hydroxycarbofuran 4. Propoxur 5. Carbaryl 6. Methiocarb
Column: Platinum™ EPS C18, 5µm, 250 x 4.6mm (Part No. 32246)
Mobile Phase: A: DI water B: Acetonitrile
Gradient: Time: 0 5 20 25 30
%B: 25 25 50 50 25
Flow Rate: 1mL/min
Detector: UV at 210nm
76www.discoverysciences.com
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spe applications
Before SPE Clean-up
After SPE Clean-up
0 10 20 30 35 405 15 25 45 50 Min.
1
23
4
56 7 8
910
11
1213
151416
Before SPE Clean-up
After SPE Clean-up
0 10 20 30 35 405 15 25 45 50 Min.
1
23
456 7
8
109 1211 13
1514
16
PAHs in Water2771
2772
PAHs in Toasted Oat Cereal
2775
2791
1. Naphthalene 2. Acenaphthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene
9. Benzo(a)anthracene 10. Chrysene 11. Benzo(b)fluoranthene 12. Benzo(k)fluoranthene 13. Benzo(a)pyrene 14. Indeno(1,2,3-cd)pyrene 15. Dibenzo(a,h)anthracene 16. Benzo(ghi)perylene
Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)
Temp: 80°C (1 min hold) to 160°C (0 min hold) at 25°C/min,
then to 300°C (0 min hold) at 3°C/min, then to 325°C
(3 min hold) at 25°C/min
Carrier Gas: Helium at 1mL/min (28cm/sec)
Detector: FID at 340°C
1. Naphthalene 2. Acenaphthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene
9. Benzo(a)anthracene 10. Chrysene 11. Benzo(b)fluoranthene 12. Benzo(k)fluoranthene 13. Benzo(a)pyrene 14. Indeno(1,2,3-cd)pyrene 15. Dibenzo(a,h)anthracene 16. Benzo(ghi)perylene
Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)
Temp: 80°C (1 min hold) to 160°C (0 min hold) at 25°C/min,
then to 300°C (0 min hold) at 3°C/min, then to 325°C
(3 min hold) at 25°C/min
Carrier Gas: Helium at 1mL/min (28cm/sec)
Detector: FID at 340°C
PAHs
Procedure using GracePure™ C18-Max:
Sample Treatment – 500mL of water and add 75mL of 2-propanol,
spiked with 20uL of PAH mixture at 2000ug/mL of each PAH, which corresponds to
40ug/500mL water, or 80ppb of each PAH.
Conditioning – 2 x 5mL of methanol, then 5mL of 85% water/
15% 2-propanol.
Sample Application – Attach 75mL reservoir (Part No. 210575) with a
syringe adapter (Part No. 210705) and apply entire sample. Dry
columns for 30 minutes at full vacuum.
Wash – No wash.
Elution – Elute PAHs with 3 x 1mL CH2Cl2 and adjusted the volume
to 1mL by evaporating under a gentle nitrogen stream.
Procedure using GracePure™ C18-Max:
Sample Treatment – 500mL of water and add 75mL of 2-propanol, spiked with 20uL of
PAH mixture at 2000ug/mL of each PAH, which corresponds to 40ug/500mL water, or
80ppb of each PAH.
Conditioning – 2 x 5mL of methanol, then 5mL of 85% water/15% 2-propanol.
Sample Application – Attach 75mL reservoir (Part No. 210575) with a syringe adapter
(Part No. 210705) and apply entire sample. Dry columns for 30 minutes at full vacuum.
Wash – 2 x 500µL of 85% water/15% isopropyl alcohol. Vacuum 10 min.
Elution – Elute PAHs with 3 x 1mL CH2Cl2 and adjusted the volume to 1mL by
evaporating under a gentle nitrogen stream.
77 www.discoverysciences.com
environmental
spe applications
0 5 10 15 20 25 30 35 Min.
Before SPE
Concentration
1
2
3
4
56
After SPE
Concentration
0 5 10 15 20 25 30 35 40 45 50 55 Min.
1
2
3 4 56 7
89 10
1112
13
14
1516
Before SPE
Concentration
After SPE
Concentration
PAHs in Cooking Oil2776
2792
1. Naphthalene 2. Acenaphthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene
9. Benzo(a)anthracene 10. Chrysene 11. Benzo(b)fluoranthene 12. Benzo(k)fluoranthene 13. Benzo(a)pyrene 14. Indeno(1,2,3-cd)pyrene 15. Dibenzo(a,h)anthracene 16. Benzo(ghi)perylene
Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)
Temp: 80°C (1 min hold) to 160°C (0 min hold) at 25°C/min,
then to 300°C (0 min hold) at 3°C/min, then to 325°C
(3 min hold) at 25°C/min
Carrier Gas: Helium at 1mL/min (28cm/sec)
Detector: FID at 340°C
PAHs
Nitroaromatics
10597
10598
Nitroaromatics and Naphthols from Soil
Column: Adsorbosphere™ UHS C18, 5µm, 150m x 4.6mm
(Part No. 288118)
Mobile Phase: Methanol:Water (50:50)
Flow Rate: 1.0mL/min
Detector: UV at 254nm
Procedure using GracePure™ C18-Max, 500mg:
Sample Treatment – Spike 100g soil with 7.5µg/g of each analyte. Combine soil with
1000mL deionized water, shake for 10 minutes, and filter.
Conditioning – Rinse device with 5mL methanol followed by 5mL water.
Sample Application – Aspirate 1000mL water sample through SPE at flow rate 1-
5mL/min.
Wash – No wash. Air dry for 15 seconds.
Elution – Elute with three 1mL aliquots of methanol:water (50:50). Air dry
for 15 seconds between each elution.
1. 2,4-Dinitrotoluene 2. 1-Naphthol 3. 2-Naphthol 4. 2-Nitrotoluene 5. 4-Nitrotoluene 6. 3-Nitrotoluene
Procedure using DVB Extract-Clean™, 500mg/4mL:
Sample Treatment – Weigh out 1 grams canola oil, spike with 20µg of each PAH, add
10mL n-hexane.
Conditioning – 10mL toluene, applied vacuum until toluene removed, then 5mL n-hexane.
Sample Application – Aspirate the entire canola oil/hexane/PAH solution through the tube.
Wash – 5mL n-hexane.
Elution – Elute PAHs with 4 X 2.5mL toluene and adjusted the sample volume to 0.5mL
(500µL) by evaporating under dry nitrogen stream and gentle heat.
78www.discoverysciences.com
enviro
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spe applications
0 2 4 6 8 10 12 14 16 18 20 Min.
Before SPE Clean-up
After SPE Clean-up
1.0
1.2
1.4
1.6
1.8
0.0
0.2
0.4
0.6
0.8
2.0
2.2
2.4
2.8
2.6
3.0
1.0
1.2
1.4
1.6
1.8
0.0
0.2
0.4
0.6
0.8
2.0
2.2
2.4
2.8
2.6
3.0
Resp
on
se -
Mil
liV
olt
s
1
2
34
5
1. Simazine
2. Atrazine
3. Propazine
4. Terbuthylazine
5. Cyanazine
GC Conditions
Injected 1.6uL of above sample onto AT™ 5ms column, 30m x 0.25mm i.d., 0.25μm film thickness (Part #15807) under the following conditions:
Oven: 80°C (0 minutes)-280°C (0 minutes) at 10°C/min
Injector: 250°C
FID: 310°C, 14.5psig helium
Linear Velocity: 29cm/sec
Flow: 1.1mL/minute
Split Ratio: 50:1
SVF: 55mL/minute (all per Agilent 6890)
SPE Conditions
Procedure using GracePure™ C18-Fast columns, 500mg/3mL, (Part #5138758)
Sample Treatment: Spiked 500mL of water
spiked with 10ug each triazine herbicide
(simazine, atrazine, propazine, terbuthylazine,
and cyanazine).
Conditioning: 2 x 2mL acetone, 2 x 2mL
methanol, 2 x 2mL water. Column must not
run dry!
Sample Application: Attached a 75mL
reservoir (Part #210575) to the SPE tube with
a syringe adapter (Part # 210705) and aspirate the sample through the tube.
Elution: Elute triazine herbicides with 3 x 1mL
acetone. Adjust sample volume to 1mL by evaporating off acetone with a warm nitrogen
stream.
Triazine Herbicides in Water
Triazine
79 www.discoverysciences.com
environmental
spe applications
GRACE® is a trademark, registered in the United States and /or other countries, of W. R. Grace & Co.-Conn.
GRACEPURE™ and GRACESMART™ are trademarks, registered in the United States and / or other countries, of
W. R. Grace & Co.-Conn. ALLTECH®, ALLTIMA®, DENALI®, EVEREST®,GENESIS®, HELIFLEX®,
VISIONHT® and VYDAC® are trademarks, registered in the United States and/or other countries,
of Alltech Associates, Inc. ADSORBOSPHERE™, ALLSEP™, ALLSPHERE™, AT™, EXPEDITE™,
EXTRACT-CLEAN™, NOVOSEP™, PLATINUM™, PREVAIL™, PROSPHERE™, PROZAP™ and ROCKET™
are trademarks, registered in the United States and/or other countries of Alltech Associates, Inc.
ASTECH™ is a trademark, registered in the United States and/or other countries, of ATEX Explosionschutz GmbH.
CHIROBIOTIC® is a trademark, registered in the United States and/or other countries, of Sigma-Aldrich
Biotechnology LP.
Conquistador® is a trademark, registered in the United States and/or other countries, of Heaven Hill Distilleries, Inc.
FLORISIL® is a trademark, registered in the United States and/or other countries, of U.S. Silica Company.
Hamilton® is a trademark, registered in the United States and/or other countries, of Hamilton Company.
ICSep™ is a trademark, registered in the United States and/or other countries, of Transgenomic, Inc.
Kellogg’s Frosted Flakes®, is a trademark, registered in the United States and/or other countries, of Kellogg North
America Company.
Pepsi One® is a trademark, registered in the United States and/or other countries, of PepsiCo, Inc.
ROBITUSSIN® is a trademark, registered in the United States and/or other countries, of Wyeth LLC.
SaraSep™ is a trademark, registered in the United States and/or other countries, OF Transgenomic, Inc.
SPLENDA® is a trademark, registered in the United States and/or other countries, of McNeil Nutritionals, Inc.
Taxol® is a trademark, registered in the United States and/or other countries, of Bristol-Myers Squibb Company.
ZirChrom® is a trademark, registered in the United States and/or other countries, of Zirchrom Separations, Inc.
This catalog is an independent publication and is not affiliated with, nor has it been authorized, sponsored,
or otherwise approved by any of the companies referenced above. This trademark list has been compiled using
available published information as of the publication date of this brochure and may not accurately reflect current
trademark ownership or status. ©Copyright 2015 Alltech Associates, Inc. All rights reserved.
grace.com
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