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Agilent New 6470 Triple Quadrupole LC/MS System
ROCK SOLID Performance For Confident Quantitation
Na Pi Parra, Ph.D
Product Manager for Triple Quadrupole LC/MS
6/23/2015
ASMS 2015 Breakfast Seminar
1
Overview of Topics
The New 6470 QQQ LC/MS System
• Technology innovations
• Quantitation performance
Key Applications
• Food safety
• Environmental
• Pharmaceutical
Summary
6/23/2015
ASMS 2015 Breakfast Seminar
2
Introducing the 6470 Triple Quadrupole LC/MS
• Rock Solid Performance
• Improved Sensitivity
• Lower IDL < 4.0 fg
• Smaller Footprint
• Upgradeable to 6495
6/23/2015
ASMS 2015 Breakfast Seminar
3
Higher Throughput
Higher Confidence
Higher Productivity
64606470
6/23/2015
ASMS 2015 Breakfast Seminar
4
6470 QQQ TechnologiesEnhanced Performance in a Smaller Space
Agilent Jet Stream Technology
• Thermal gradient focusing
• Efficient desolvation
• Creates an ion rich zone
• Up to 10x gains in sensitivity
• An Ion Detector with High Energy Conversion Dynode and Low Noise
• Improved ion detection3
• A Curved and Tapered Hexapole Collision Cell
• Effective ion collection2
• Enhanced Q1 Ion Optics
• Improved ion transmission1
Enhanced Q1 Ion Optics with Optimized PrefilterImproved transmission of precursor ions and additional system robustness
6/23/2015
ASMS 2015 Breakfast Seminar
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1. Reduced Octopole length1
1. New prefilter2
1
2
• New optimized MS 1 prefilter geometry for improved precursor ion transmission
– Improved peak area response and peak area %RSD, more sensitive and precise
• New optical lens elements for reduced the probability of contamination
– More reliable and robust performance
Curved and Tapered Hexapole Collision Cell Effective collection and transmission of product ions and smaller footprint
6/23/2015
ASMS 2015 Breakfast Seminar
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• Curved and Tapered Hexapole Assembly for efficient collection and transmission of
product ions
• Designed for consistent collision energies across all QQQ platforms
• A compact, smaller benchtop footprint
6460 QQQ LC/MS
- Linear Collision Cell
6470 QQQ LC/MS
- Curved Collision Cell
30% smaller
New Detector with High Energy Conversion DynodeMore efficient detection and quantitation of ions with low noise characteristics
6/23/2015
ASMS 2015 Breakfast Seminar
7
• Improved ion detection efficiency with High Energy Dynode (HED) voltage up to 20 kV
- Improved peak area response and peak area %RSD in positive & negative ion mode
- Improved sensitivity and precision for a wide mass range
• Low noise level at 20 kV
- Improved signal to noise
40 – 113% increase in
response to 20kV vs. 10kV
6470 QQQ – Rock Solid, Improved PerformanceRobust and Reliable, More Sensitive, More Compact, and Upgradeable
6/23/2015
ASMS 2015 Breakfast Seminar
8
Proven robustness and reliability in complex matrix
- Extensively tested in key applications
Improved sensitivity and lower LLOQs
– In S/N specifications and key applications
Improved precision and excellent accuracy
– In IDL specifications and key applications
Improved scan and MRM speed, and polarity switching
Proven 6 orders of linear dynamic range
Smaller (30%) footprint and upgradeable to the 6495
Per
form
ance
Robustness
Footprint
Unprecedented Robustness in Biological MatrixQuantitation of Drugs in Plasma using RapidFire 365 + 6470 QQQ
6/23/2015
ASMS 2015 Breakfast Seminar
9
• Stable peak area response over 10,000 injections of plasma samples and 5 days of
continuous operation
• Response %RSD = 3.7 – 8.6
Injection #1
Injection #10,000
Injection #1
Injection #10,000
Time (Sec)
Time (Sec)
Love et al.
WP #496
Improved Sensitivity and Precision – Lower IDLsReserpine (+) and Chloramphenicol (-), 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
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10 fg reserpine (+) on the 6470 10 fg chloramphenicol (-) on the 6470
IDL = 2.5 fg %RSD = 8.7
n = 10 injections
%RSD = 10.2
n = 10 injections
IDL = 2.9 fg
Models Reserpine (+) Chloramphenicol (-)
Measured at %RSD IDL Measured at %RSD IDL
6460 20 fg 9.6 5.4 fg 20 fg 11.9 6.7 fg
6470 10 fg 10.2 2.9 fg 10 fg 8.7 2.5 fg
6470 achieves 2-3x lower IDLs vs. the 64606470 IDLs are measured at a 2x lower level
Improved Sensitivity and Precision – Lower LLOQAcetaminophen LLOQ in Water, 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
11
• Improved peak area (4x) and area %RSD (2x) were observed on the 6470 compared to the 6460
• Lower LLOQ (4x) was observed on the 6470 compared to the 6460
• Excellent precision (%RSD) and accuracy were achieved at the LLOQ level of 6470
LLOQ = 2 ng/L on the 6460
6470
Peak Area = 5863
S/N = 34.6
Area %RSD = 4.3
6460
Peak Area = 1490
S/N = 11.1
Area %RSD = 9.5
n = 8 injections
LLOQ = 0.5 ng/L on the 6470
Blank
6470
Peak Area = 1656
S/N = 10.6
Area %RSD = 8.3
%Accuracy = 104.8
6460
Peak Area = 423
S/N = 4.0
Area %RSD = 19.0
% Accuracy = 77.0
n = 8 injections
Excellent Precision at the Lowest Levels – Lower IDLAcetaminophen IDL measured at the LLOQ level, 6470 vs. 6460
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ASMS 2015 Breakfast Seminar
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Models Amount measured - at LLOQ Replicates %RSD IDL
6460 2 ng/L n = 8 9.5 0.59 ng/L
6470 0.5 ng/L n = 8 8.3 0.12 ng/L
• Lower LLOQ (4x) and IDL (5x) were achieved on
the 6470 compared to the 6460
Inj # Peak Area S/N
1 1435 22.2
2 1419 10.6
3 1373 11.0
4 1282 10.6
5 1343 7.6
6 1431 15.5
7 1656 15.2
8 1553 10.6
%RSD 8.3 Ave 12.8
0.5 ng/L
0.5 ng/L
0.5 ng/L
0.5 ng/L
0.5 ng/L
0.5 ng/L
0.5 ng/L
0.5 ng/L
Acetaminophen
6470 IDL = 0.1 ng/L
6460 IDL = 0.5 ng/L
Blank
Attogram Sensitivity (+)500 attograms of Clenbuterol on-column
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ASMS 2015 Breakfast Seminar
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LLOQ of Clenbuterol on the 6470
500 attogram on-column
Blank
Area %RSD = 12.4
n = 3 injections
%Accuracy = 107.2
S/N = 5.9
Excellent precision (%RSD) and accuracy are observed at the LLOQ level of 6470
Attogram Sensitivity (-)800 attograms of Chloramphenicol on-column
6/23/2015
ASMS 2015 Breakfast Seminar
14
Area %RSD = 18.1
n = 3 injections
%Accuracy = 98.2
S/N = 18.0
Excellent precision (%RSD) and accuracy are observed at the LLOQ level of 6470
LLOQ of Chloramphenicol on the 6470
800 attogram on-column
Blank
5.7 Orders of Linear Dynamic Range Clenbuterol (+), 0.1 pg/mL – 50 ng/mL
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ASMS 2015 Breakfast Seminar
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• Excellent linearity, R2 = 0.9995
• Superb assay accuracy and precision are achieved at all
levels including the LLOQ level of 0.1 pg/mL
Levels%RSD
(n = 3)
%
Accuracy
0.1 pg/mL 12.4 107.2
0.5 pg/mL 9.4 100.2
1 pg/mL 4.4 81.0
5 pg/mL 1.7 81.1
50 pg/mL 0.36 83.0
500 pg/mL 2.0 88.2
5 ng/mL 0.60 105.6
50 ng/mL 0.20 99.6
Applications for 6470 QQQ LC/MS
6/23/2015
ASMS 2015 Breakfast Seminar
16
Food Safety
Analysis of multi-residue pesticides in food matrices with higher degrees of sample dilution
Environmental
Determination of PPCPs in surface water using direct injection
Pharmaceutical – High-Capacity Multisampler UHPLC
High-throughput analysis of drugs & metabolites in plasma with reliable 24/7 operation
Multi-Residue Pesticide Analysis in Food
6/23/2015
ASMS 2015 Breakfast Seminar
17
• Multi-residue pesticide analysis in food products – most demanding food safety applications
• Improved sensitivity and precision of the 6470 allows accurate quantitation of pesticides
<Maximum Residue Limits (MRLs) imposed by EU, with higher degrees of sample dilution
• Sample dilution reduces matrix effects, improves method robustness, allows more efficient
ionization and enables the use of solvent calibration with better accuracy
Injection volume = 2 µL
1:20 dilution of > 250 pesticides spiked into black tea at 10 ng/mL (ppb)
Sartain et al.
WP #059
Sensitivity Improvements for High Relevance Pesticides1:20 Dilution of 10 ng/mL of Pesticides in Black Tea, Peak Area 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
18
• Improved sensitivity (average peak area gain of 3.0x) are observed on the new 6470 vs. 6460
6470 6460
Oxamyl
2.8x
Methomyl
3.9x
Area Gain
Thiamethoxam
2.5x
6470 6460 Area Gain
Acetamiprid
3.4x
Thiacloprid
2.7x
Ethion
3.2x
Improved Precision and Lower IDLsIDLs of 30 Highly Relevant Pesticides, 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
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• Improved precision (peak area %RSD) are observed, particularly at the lowest levels
• The enhanced peak area response and improved precision (%RSD) leads to lower
LLOQs and IDLs compared to the 6460
• Lower IDLs (average improvement of 3.6x) was observed across all > 250 pesticides
Pesticide IDL (ppt) Fold-
Improvement
Pesticide IDL (ppt) Fold-
Improvement 6470 6460 6470 6460
Acephate 7.1 10.4 1.5x Fipronil 74.5 339.7 4.6x
Acetamiprid 1.8 32.9 18.0x Flufenoxuron 58.0 521.6 9.0x
Azinphos-methyl 165.1 278.9 1.7x Imazalil 30.1 203.2 6.7x
Buprofezin 6.2 36.0 5.8x Imidacloprid 57.6 191.1 3.3x
Carbendazim 3.7 13.5 3.6x Isocarbophos 14.6 129.5 8.9x
Chloroxuron 7.4 98.4 13.3x Metamitron 172.6 558.4 3.2x
Cycluron 8.4 71.1 8.4x Methidathion 71.9 340.1 4.7x
Desmedipham 11.2 32.4 2.9x Methomyl 4.1 21.5 5.2x
Diethofencarb 11.1 59.0 5.3x Monocrotophos 10.8 52.3 4.8x
Dimethoate 2.8 32.8 11.8x Oxamyl 0.9 5.8 6.2x
Dimethomorph 59.0 310.7 5.3x Pirimicarb 0.8 2.8 3.6x
Dimoxystrobin 6.3 23.6 3.8x Pyridaben 0.8 0.6 0.8x
Diniconazole 30.9 237.4 7.7x Tebuconazole 45.8 41.9 0.9x
Dioxacarb 103.7 139.1 1.3x Teflubenzuron 291.6 2012.1 6.9x
Diuron 22.6 37.4 1.7x Triazophos 10.1 65.9 6.6x
Average Improvement 3.6X
Femtogram Sensitivity for Pesticides in Black TeaLLOQs and IDLs / MDLs at Low-pg/mL level on the 6470
6/23/2015
ASMS 2015 Breakfast Seminar
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• LLOQs 100 pg/mL are required to achieve the MRL with 1:20 dilution of black tea
• Excellent peak area precision (%RSD) and accuracy are observed at the LLOQs in black tea
Pesticides
Oxamyl (+)
Pirimicarb (+)
Bentazone (-)
Flubendiamide (-)
Black Tea
LLOQ
(pg/mL)%Accuracy
%RSD
(n = 5)
MDL
(pg/mL)
10 108.9 9.5 3.6
10 103.7 10.4 3.9
10 98.2 19.2 7.2
20 105.4 17.8 13.3
Neat Solvent
LLOQ
(pg/mL)%Accuracy
%RSD
(n = 5)
IDL
(pg/mL)
2 114.0 12.6 0.94
2 117.2 10.3 0.77
5 81.8 16.3 3.1
10 86.0 12.4 4.7
Bentazone 10 pg/mL
S/N = 17.6
Oxamyl 10 pg/mL
S/N = 10.4
Flubendiamide 20 pg/mL
S/N = 195Pirimicarb 10 pg/mL
S/N = 12.7
LLOQs in Black Tea
Improved Sensitivity and Precision of the 6470 Allows for Higher Degree of Sample Dilution
6/23/2015
ASMS 2015 Breakfast Seminar
21
• Majority of the pesticides achieved an LLOQ of 5% of the MRL level in food matrices
• Pesticides can be confidently quantified in food matrices with up to 1:20 dilution
• Sample dilution leads to reduced matrix effects and improved method robustness
• Sample dilution allows for more efficient ionization and significantly better recoveries
Histograms of LLOQs for Evaluated Pesticides in Solvent and Food Matrices
≤ 100 ppt 100-200 ppt 200-500 ppt 500 ppt - 1 ppb 1-10 ppb >10 ppb
171
3226
10 9 11
163
3826
9 13 10
101
3446
27 3121
Tomato Orange Black Tea
Improved Recoveries at Higher Degree of Dilution
6/23/2015
ASMS 2015 Breakfast Seminar
22
0
10
20
30
40
50
60
70
80
90
100
<30% 30-50% 50-70% 70-120% >120%
Num
ber o
f Pes
ticid
es
Recoveries
undiluted
1 to 2
1 to 5
1 to 10
1 to 20
PassesSANCO
Requirements
• 116 pesticides showed strong ion suppression in undiluted black tea
• Most of pesticides achieved acceptable recoveries of 70-120% and less signal
suppression with 1:20 dilution, which are in full compliance with SANCO requirements
• Neat solvent calibration curve can be used for pesticide quantitation with better accuracies
Applications for 6470 QQQ LC/MS
6/23/2015
ASMS 2015 Breakfast Seminar
23
Food Safety
Analysis of multi-residue pesticides in food matrices with higher degrees of sample dilution
Environmental
Determination of PPCPs in surface water using direct injection
Pharmaceutical – High-Capacity Multisampler UHPLC
High-throughput analysis of drugs & metabolites in plasma with reliable 24/7 operation
Determination of PPCPs in Environmental Water
6/23/2015
ASMS 2015 Breakfast Seminar
24
• PPCPs in surface waters may have an adverse impact on wildlife and human even at very low
concentrations (ng/L)
• The improved sensitivity and precision of the 6470 allows accurate quantitation of PPCPs at
sub-ng/L level using direct injection without the need of time-consuming SPE enrichment
• Direct injection improves analysis efficiency, assay accuracy, and lab productivity
28 PPCPs spiked in water at 10 ng/L (ppt)
Injection volume = 40 µL
24X IN POS Mode Erythromycin (+)
Acetaminophen (+) Fluridone (+)
Atenolol (+) Gabapentin (+)
Atrazine (+) Lamotrigine (+)
Bupropion (+) Metoprolol (+)
Caffeine (+) Propranolol (+)
Carbamazepine (+) Sucralose (+)
Clarithromycin (+) Sulfamethoxazole (+)
Cotinine (+) Trimethoprim (+)
DEET (+) Venlafaxine (+)
Dextrorphan (+) 4x in NEG Mode
Diazinon (+) 2,4-D (-)
Diltiazem (+) Gemfibrozil (-)
Diphenhydramine (+) Triclopyr (-)
Diuron (+) Triclosan (-)
Li et al.
TP #305
Improved Sensitivity for PPCPs in Positive Mode10 ng/L of PPCPs in Water, Peak Area 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
25
Acetaminophen 2.6x
Atenolol 2.3x
Atrazine 3.3x
Bupropion 3.5x
Caffeine 2.4x
Carbamazepine 2.2x
Clarithromycin 4.5x
Cotinine 2.9x
DEET 2.6x
Dextrorphan 1.8x
Diazinon 2.6x
Diltiazem 2.1x
• Improved sensitivity (average peak area gain of 2.9x) are observed across 24 PPCPs in POS mode
6470
6460
Improved Sensitivity for PPCPs in Negative Mode10 ng/L of PPCPs in Water, Peak Area 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
26
2, 4-D 4.1x Triclopyr 3.8xGemfibrozil 2.3x Triclosan 2.7x
• Improved sensitivity (average peak area gain of 3.2x) are observed across 4 PPCPs in NEG mode
10 ppt on 6460
10 ppt on 6470
2.9x in POS compounds
3.2x for NEG compounds
Average improvement = 3.0x
Overlaid MRM chromatograms: 6470 vs. 6460
6470
6460
Improved Precision – Comparison of Area %RSD5 ng/L of PPCPs in Water, 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
27
• Improved precision (peak area %RSD) are observed, particularly at the lowest levels
• Average improvement of 2.3x was observed across 28 PPCPs analyzed
• The enhanced peak area response and improved precision (%RSD) leads to lower IDLs and
LLOQs compared to the 6460
Compounds%RSD (n=8) Improvement
Compounds%RSD (n=8)Improvement
Acetaminophen 0.6x Erythromycin 1.3x
Atenolol 1.6x Fluridone 6.0x
Atrazine 2.0x Gabapentin 1.7x
Bupropion 2.7x Lamotrigine 3.3x
Caffeine 3.6x Metoprolol 1.3x
Carbamazepine 4.9x Propranolol 1.8x
Clarithromycin 1.3x Sucralose 1.2x
Cotinine 3.0x Sulfamethoxazole 1.6x
DEET 2.2x Trimethoprim 2.1x
Dextrorphan 1.8x Venlafaxine 2.0x
Diazinon 2.0x 2,4-D 2.1x
Diltiazem 1.4x Gemfibrozil 3.6x
Diphenhydramine 4.0x Triclopyr 2.4x
Diuron 1.0x Triclosan 1.2x
Average Improvement 2.3x
Lower IDLs at Sub-ng/L Level IDLs of PPCPs in water, 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
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• Lower IDLs (average improvement of 3.6x) were observed across all 28 PPCPs (POS and NEG)
PPCPs6460 IDL
(ng/L)
6470 IDL
(ng/L)Improve
mentPPCPs
6460 IDL
(ng/L)
6470 IDL
(ng/L)Improve
ment
Acetaminophen 0.59 0.12 4.9x Erythromycin 0.96 0.14 6.8x
Atenolol 0.64 0.45 1.4x Fluridone 0.053 0.012 4.5x
Atrazine 0.44 0.28 1.6x Gabapentin 1.89 1.05 1.8x
Bupropion 0.17 0.044 3.9x Lamotrigine 2.15 0.94 2.3x
Caffeine 1.20 0.25 4.8x Metoprolol 1.02 0.29 3.5x
Carbamazepine 0.32 0.082 3.9x Propranolol 0.75 0.091 8.2x
Clarithromycin 3.58 1.14 3.1x Sucralose 19.5 3.56 5.5x
Cotinine 0.57 0.068 8.0x Sulfamethoxazole 0.87 0.41 2.1x
DEET 0.067 0.022 3.1x Trimethoprim 0.64 0.39 1.6x
Dextrorphan 1.01 0.15 6.7x Venlafaxine 0.22 0.076 2.8x
Diazinon 0.17 0.071 2.4x 2,4-D 34.4 8.69 4.0x
Diltiazem 0.13 0.030 4.2x Gemfibrozil 22.1 11.5 1.9x
Diphenhydramine 0.13 0.052 2.4x Triclopyr 38.0 16.3 2.3x
Diuron 0.56 0.28 2.0x Triclosan 14.6 5.32 2.7x
Average Improvement 3.6x
IDLs of 24 PPCPs in Positive Mode
IDLs of 4 PPCPs in Negative Mode
6470 IDL 6460 IDL
6470 IDL 6460 IDL
Lower LLOQs – Femtogram Sensitivity for PPCPsLLOQs of PPCPs in water, 6470 vs. 6460
6/23/2015
ASMS 2015 Breakfast Seminar
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• Lower LLOQs (average improvement of 2.7x) were achieved across all 28 PPCPs (POS and NEG)
• LLOQs for most of the PPCPs are much lower than the ng/L (ppt) level regulation limit
• Excellent peak area precision (%RSD) and accuracy are observed at the LLOQs
PPCPs 6470 LLOQ
(ng/L)
6460 LLOQ
(ng/L) PPCPs
6470 LLOQ
(ng/L)
6460 LLOQ
(ng/L)
Acetaminophen 0.5 2 Erythromycin 1.0 2
Atenolol 1.0 2 Fluridone < 0.1 0.5
Atrazine 0.5 2 Gabapentin 5.0 10
Bupropion 0.2 1 Lamotrigine 2.0 10
Caffeine 1.0 2 Metoprolol 1.0 2
Carbamazepine 0.5 2 Propranolol 1.0 2
Clarithromycin 5.0 10 Sucralose 20.0 50
Cotinine 0.5 2 Sulfamethoxazole 1.0 2
DEET 0.2 0.5 Trimethoprim 1.0 2
Dextrorphan 1.0 2 Venlafaxine 0.5 1
Diazinon 0.5 1 2,4-D 20.0 50
Diltiazem 0.2 0.5 Gemfibrozil 20.0 50
Diphenhydramine 0.2 0.5 Triclopyr 50.0 100
Diuron 1.0 2 Triclosan 20.0 50
Improvement 2 – 5 X
%RSD = 3.8 (n=8)%Accuracy = 102.3S/N = 66
%RSD = 8.3 (n=8)%Accuracy = 104.8S/N = 22
%RSD = 7.3 (n=8)%Accuracy = 99.0S/N = 35.1
%RSD = 8.9 (n=8)%Accuracy = 107.9S/N = 33.4
Acetaminophen 0.5 ng/L Fluridone 0.1 ng/L
Bupropion 0.2 ng/L Triclosan 20 ng/L
Applications for 6470 QQQ LC/MS
6/23/2015
ASMS 2015 Breakfast Seminar
30
Food Safety
Analysis of multi-residue pesticides in food matrices with higher degrees of sample dilution
Environmental
Determination of PPCPs in surface water using direct injection
Pharmaceutical – High-Capacity Multisampler UHPLC
High-throughput analysis of drugs & metabolites in plasma with reliable 24/7 operation
Analysis of Drugs and Metabolites in Human Plasma6470 QQQ with High-Capacity Multisampler UHPLC
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ASMS 2015 Breakfast Seminar
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• PK analysis of drugs and metabolites is challenged with the need to perform large-scale-analyses
with high sensitivity (pM) and sufficient dynamic range (pM nM) in biological matrices
• The highly sensitive 6470 enables fast and accurate quantitation of drugs and metabolites at
low-pM level over a wide linear dynamic range in human plasma
• 6470’s robustness together with the high-capacity multisampler allows for high-throughput drug
analysis with 24/7 unattended operation and extremely reliable results
-Hydroxymidazolam
Midazolam
Alprazolam
- Hydroxyalprazolam
-OH MidazolamMidazolam
Alprazolam -OH Alprazolam
Benzodiazepines and metabolites spiked in human plasma at 5 pg/mL
Yang et al.
TP #297
High Sensitivity for Benzodiazepines in Plasma LLOQs and MDLs at Low-pg/mL level on the 6470
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• Excellent peak area precision (%RSD) and accuracy are observed at the LLOQ level
-Hydroxymidazolam
LLOQ = 5 pg/mL
S/N = 13.6
Midazolam
LLOQ = 5 pg/mL
S/N = 35.2
-Hydroxyalprazolam
LLOQ = 5 pg/mL
S/N = 5.4
Alprazolam
LLOQ = 5 pg/mL
S/N = 9.2
CompoundLLOQ
(pg/mL)%Accuracy
%RSD
(n = 7)
MDL
(pg/mL)
-Hydroxymidazolam 5 100.7 6.9 1.1
Midazolam 5 114.6 5.3 0.84
-Hydroxyalprazolam 5 99.4 9.9 1.6
Alprazolam 5 96.1 7.5 1.2
Confident Quantitation over 5 Orders of Dynamic Range
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• Excellent linearity (R2 > 0.995), accuracy and precision are achieved at all levels including LLOQ
• Drugs and metabolites at different levels can be quantified simultaneously and in one experiment
Levels %RSD (n = 7) %Accuracy
5 pg/mL 5.32 114.6
25 pg/mL 2.76 96.6
50 pg/mL 2.53 102.8
250 pg/mL 1.22 98.6
500 pg/mL 0.73 98.7
2.5 ng/mL 0.51 97.6
5 ng/mL 0.34 99.1
25 ng/mL 0.56 96.5
50 ng/mL 0.79 96.4
250 ng/mL 0.75 97.3
500 ng/mL 0.47 101.9
Midazolam, 5 pg/mL – 500 ng/mL
5 Orders of Linear Dynamic Range
R2 = 0.9994
Concentration (ng/ml)0.005 0.01 0.05 0.1 0.5 1 5 10 50 100 500 1000
Rela
tive
Res p
ons e
s 2x10
5E-06
1E-05
5E-05
0.0001
0.0005
0.001
0.005
0.01
0.05
0.1
0.5
Levels %RSD (n = 7) %Accuracy
5 pg/mL 7.54 96.1
25 pg/mL 2.77 96.9
50 pg/mL 2.95 101.9
250 pg/mL 1.62 101.3
500 pg/mL 1.20 101.8
2.5 ng/mL 0.42 99.8
5 ng/mL 1.24 102.3
25 ng/mL 2.15 100.1
50 ng/mL 0.77 99.7
250 ng/mL 2.91 100.0
500 ng/mL 0.62 86.6
Alprazolam, 5 pg/mL – 500 ng/mL
5 Orders of Linear Dynamic Range
R2 = 0.9970
Concentration (ng/ml)0.005 0.01 0.05 0.1 0.5 1 5 10 50 100 500 1000
Rela
tive
Res p
ons e
s 2x10
5E-06
1E-05
5E-05
0.0001
0.0005
0.001
0.005
0.01
0.05
0.1
0.5
Proven System Robustness in Human Plasma
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• Stable peak area response was observed over 1,300 injections of plasma extract and 1 week
of continuous operation
• Average area ratio %RSD = 1.8 was observed with use of internal standard (ISTD)
• Average peak area %RSD = 6.6 was observed for benzodiazepines and metabolites
0
0.2
0.4
0.6
0.8
1
1.2
0 200 400 600 800 1000 1200 1400
Peak A
rea R
ati
o
Injection number
-Hydroxymidazolam
0
0.2
0.4
0.6
0.8
1
1.2
0 200 400 600 800 1000 1200 1400
Peak A
rea R
ati
o
Injection number
Midazolam
0
0.2
0.4
0.6
0.8
1
1.2
0 200 400 600 800 1000 1200 1400
Peak A
rea R
ati
o
Injection number
-Hydroxyalprazolam
0
0.2
0.4
0.6
0.8
1
1.2
0 200 400 600 800 1000 1200 1400P
eak A
rea R
ati
oInjection number
Alprazolam
Area Ratio %RSD=1.3
Peak area %RSD=5.7
Area Ratio %RSD=3.3
Peak area %RSD=7.0
Area Ratio %RSD=1.4
Peak area %RSD=7.2
Area Ratio %RSD=1.3
Peak area %RSD=6.3
6/23/2015
35
Summary: 6470 QQQ LC/MSRock Solid Performance for Confident Quantitation and Highest Lab Productivity
Additional Robustness
Improved Sensitivity
Improved Precision
Faster Speed
Smaller Footprint
Upgradeable to the 6495
Ion Optics
20KV Detector
Curved Collision Cell
Less Maintenance Cleaning
Reliable Consistent Results
over Longer Time (24/7)
Streamlined Analytical
Workflow
Reproducible
High Quality Data
Higher Throughput
at UHPLC Speed
More Instruments
In Your Laboratory
Protection of
Your Investment
ASMS 2015 Breakfast Seminar
6/23/2015
ASMS 2015 Breakfast Seminar
36
ASMS 2015, St Louis6470 Marketing Launch
Breakfast Seminar
Technical VIP
Technical Posters
Brochure
App Note
Hospitality Suite
Agilent Booth
6/23/2015
ASMS 2015 Breakfast Seminar
37