Jing Yu Shen*, Chang-Joo Lee 1 and Jae-Han Shim

1. Determination of Enrofloxacin and Ciprofloxacin in Chicken Muscle using SFE and HPLC Jing Yu Shen*, Chang-Joo Lee1 and Jae-Han Shim Division of Applied Bioscince and Biotechnology, College of Agricultural Science and Life Science, Chonnam National University, Gwang Ju, 500-757, Korea. 1Division of Civil and Environmental Engineering, Kwangju University1, Gwang Ju, 500-703, Korea. Abstract Enrofloxacin (ENR) and ciprofloxacin (CIP) are fluoroquinolones antimicrobial agents used in animals and human. The use of fluoroquinolones has been steadily increasing to become the major antibacterial agents in livestock industry. Public concern over drug residues in our food supply and the potential for the emergence of drug resistant bacteria will generate a need for an integrated monitoring and detection system. Now the most common quantitative analysis method for fluoroquinolones is high performance liquid chromatography (HPLC) with fluorescence detector. For HPLC analysis, traditional solvent extraction is necessary and this process require multiple steps and consume large amounts of toxic organic solvents. In order to solve this problem, supercritical fluid extraction(SFE) was employed. The optimum SFE condition, limit of detection(LOD), limit of quantitation(LOQ) and recoveries were studied. Furthermore, the residues of enrofloxacin and ciprofloxacin were investigated in chicken tissue after orally administered dose of 10 mg/kg of body weight once daily for 5days. Objectives Table 1. SFE optimal condition of enrofloxacin and ciprofloxacin from fortified chicken muscle. Develop the method for antibacterial agent ENR and CIP in chicken uscle. Avoid use organic solvent extraction and separation by using SFE. Find the optimum SFE conditions (system pressure, oven temperatrue and modifier ratio) Investigated the residues of ENR and CIP after orally administration chicken muscle. Materials and Methods These chickens purchased at poultry farm(Chonnam ). ENR(Bayer, 99.2%), CIP(Cheil Jedang, 100%). CO2 (Shin Jung gas). Methanol (Merck). SFE(Supercritical fluid extractor. Jasco. Japan) HPLC (Shimadzu. Japan), Fluorescence Detector ( Wevelengths. Ex: 278nm, Em: 450nm). Column( Nova-pak C18, 3.9x300mm). Spike Sample A 2g amount of chicken muscle was transferred to a 20ml vial, 200㎕of 1ppm standard was added and freezing dried for 24 hour. Fig. 5. Muscle concentrations(mean±SD) versus time of nrofloxacing and ciprofloxacin after orally administration (10mg/kg of body weight once daily for 5day) in chicken. The optimum SFE condition were established in fortified chicken muscle. Recovery were 93.5-104.3 % for ENR and 41.9-65.3% for CIP, respectively, after oral administration. The mean residence time of ENR and CIP for chicken muscle were 120 and 48 hours, respectively. The maximal muscle concentration of ENR and CIP were 997.6 and 50.0 ㎍/kg body weight, respectively. Results and Discussion References 1. 심 재한, 우슬중 β-Ecdysone의 초임계추출법(SFE)과 용매추출법의 비교. 한국농화학회지. 2001. 2. 이 미현, SFE와 HPLC를 이용한 계란내 Fluoroquinolones 검출에 대한 연구. 전남대학교. 2002. 3. 서 계원, 계란중 Fluoroquinolone계 함성항균제의 잔류에 관한 연구. 전남대학교. 2000. 4. 손 성완, 동물성 식품내 퀴놀론계 항균물질 검출에 관한 연구. 서울대학교. 1999. 5. Pak-Sin Chu et al., Determination of total 14C residues of sarafloxacin in eggs of laying hens. J. Agric. Food Chem. 2000. 6.Ana Cabanes, DVM et al., Pharmacokinetics of enrofloxacin after intravenous and intramuscular injection in rabbites. Am. J. Vet. Res. 1999. 7. Jose J. Aramayona, PhD et al., Penetration of enrofloxacin and ciprofloxacin into breast milk, and pharmacokinetics of the drugs in lactating rabbits and neonatal offspring. Am. J. Vet. Res. 1996. 8. Nora Gorla et al., HPLC residues of enrofloxacin and ciprofloxacin in eggs of laying hens. Inte. J. Ant. Agen. 1997. 9. D. G. Kennedy et al., Use of liquid chromatography-mass spectrometry in the analysis of residues of antibiotics in meat and milk. J. chromatography A, 1998. 10. J.C. Yorke and P. Froc, Quantitation of nine quinolones in chicken tissues by high-performance liquide chromatography with fluorescence detection. J. Chromatography A, 2000. Fig. 1.Extraction profile of enrofloxacin and ciprofloxacin with various tempratrue by SFE Fig. 2. Extraction profile of enrofloxacin and ciprofloxacin with Various pressure by SFE. Fig. 3. Extraction profile of enrofloxacin and ciprofloxacin with various modifier ratio (MeOH, %) by SFE. Fig. 5. Recovery of enrofloxacin and ciprofloxacin from spiked chicken muscle by SFE. Corresponding address: http://chonnam.chonnam.ac.kr/~jhshim E-mail address: jhshim@chonnam.chonnam.ac.kr