Analysis of Residual Synthetic Antibacterials in Meat by HPLC Application

Food
Hiroki Kumagai, Adebayo Onigbinde

Many domestic cattle receive various antibacterials in their feed for the prevention and control of disease caused by fungi and bacteria. Residues of antibacterials are found in food made from the meat of these animals. Since many antibiotics are toxic, many countries regulate acceptable residue levels of compounds allowable in agricultural and animal products. Many alkyl-C18 columns tail with basic compounds and have a shorter life time at low pH. Purospher column separated basic antibacterials with good resolution, peak shape,and efficiency.
Absorbance [mAU] 25 20 15 10 5 0 0 Absorbance [mAU] 25 20 15 10 5 0 0 5

Highlights
Separation of 10 antibacterials in meat at low pH Excellent and rapid resolution of antibacterials at low sample concentration Elution of antibacterials from the column with good peak shape and narrow peak width Separation of low level amounts of a wide range of pharmaceutical compounds with differing structures in a single analysis by Purospher column

3 8 9 10
1 2 3 4 5 Time (min) SMr PYM TCP SDD FZD 6 7 8 9 10 SMMX DFZ SDMX SQX OXA

at 224 nm
1
10 15 20

4 5
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67

30

35

at 360 nm

DFZ FZD

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Time (min)

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

Absorbance [mAU] 4 3 2 1 0 -1 0 Absorbance [mAU] 4 3 2 1 0 -1 0 5 10

at 224 nm SDD

15

20

25

30

35

Time (min)

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 muscle Sample preparation: According to the official procedure of the Japanese food sanitation law.

at 360 nm

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35

Time (min)

Figure 1. Chromatogram of extract of bovine muscle.

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 min 65% 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

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Hiroki Kumagai is the PHS Support manager based at Yokogawa Analytical Systems Inc., Tokyo, Japan. Adebayo Onigbinde is an application chemist based at Agilent Technologies, Wilmington, Delaware, USA.

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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