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Many domestic cattle receive various antibacterials in their feed for the preven-tion and control of disease caused by fungi and bacteria. Residues of antibacterialsare found in food made from the meat of these animals. Since many antibioticsare toxic, many countries regulate acceptable residue levels of compoundsallowable in agricultural and animal products. Many alkyl-C18 columns tail withbasic compounds and have a shorter life time at low pH. Purospher column

separated basic antibacterials with good resolution, peak shape,and efficiency.

Analysis of Residual SyntheticAntibacterials in Meat by HPLCApplication

Food

25

20

15

10

5

0

0

Absorbance

[mAU]

5 10 15 20

FZD

DFZ

25 30 35 Time (min)

25

20

15

10

5

0

0

Absorbance

[mAU]

5 10 15 20 25 30 35 Time (min)

1 SMr

2 PYM

3 TCP

4 SDD

5 FZD

6 SMMX

7 DFZ

8 SDMX

9 SQX

10 OXA1

34

2

at 224 nm

at 360 nm

56 7

8 9 10

Hiroki Kumagai, Adebayo Onigbinde

Highlights

Separation of 10 antibacterials inmeat at low pH

Excellent and rapid resolution of

antibacterials at low sample con-centration

Elution of antibacterials from thecolumn with good peak shape andnarrow peak width

Separation of low level amounts ofa wide range of pharmaceuticalcompounds with differing struc-tures in a single analysis byPurosphercolumn

Analyzed Compounds

Sulfamerazine (SMR)

Sulfadimidine (SDD)

Sulfamonomethoxine (SMMX)

Sulfadimethoxine (SDMX

Sulfaquinoxaline (SQX)

Pyrimethamine (PYM)

Thiamphenicol (TPC)

Furazolidone (FZD)

Difurazone (DFZ)

Oxolinic acid (OXA)

Sample: Extracts from bovine muscleSample preparation: According to theofficial procedure of the Japanesefood sanitation law.

Figure 1. Chromatogram of standard solution, 2 g/mL each analyte.

Instrument: Agilent 1100 Series HPLC; Column: 250 mm 4 mm id, RP-18 Purospher, 5 m, Part no. 79925PU-584;Mobile phase: A = 0.7 % Phosphoric acid, B = CH3CN; Gradient: 0.0 min 5% B; 10.0 min 65% B; 40.0 min 65% B; 45.0 min65% B; Post Time 7.0 min 5% B; Flow rate: 1.0 mL/min; Temperature: 40 C; Injection volume: 20 L; Diode array detector:A338/10 nm, reference wavelength off; B264/8 nm, reference wavelength off; C360/8 nm, reference wavelength off

Figure 1. Chromatogram of extract of bovine muscle.

4

3

2

1

0

-1

0

Absorbance

[mAU]

5 10 15 20

SDD

25 30 35 Time (min)

4

3

21

0

-1

0

Absorbance

[mAU]

5 10 15 20 25 30 35 Time (min)

at 224 nm

at 360 nm

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Agilent shall not be liable for errors contained herein or

for incidental or consequential damages in connection

with the furnishing, performance, or use of this

material.

Information, descriptions, and specifications in this

publication are subject to change without notice.

Agilent Technologies, Inc. 2002

Printed in the USA

June 18, 2002

5988-7135EN

www.agilent.com/chem

Hiroki Kumagai is the PHS Supportmanager based at Yokogawa AnalyticalSystems Inc., Tokyo, Japan.

Adebayo Onigbinde is an applicationchemist based at Agilent Technologies,Wilmington, Delaware, USA.

For More Information

For more information on our productsand services, visit our Web site atwww.agilent.com/chem.