猪排泄物中恩诺沙星和环丙沙星含量的HPLC检测方法

doi:10.16431/j.cnki.1671-7236.2016.04.031

摘要/Abstract

摘要： 为了对猪排泄物中恩诺沙星(enrofloxacin,ENR)和环丙沙星(ciprofloxacin,CIP)进行定量检测,试验建立了测定猪粪尿中ENR和CIP含量的高效液相－荧光检测方法。将猪粪经乙腈－氨水超声提取后,加入三氯乙酸酸化,然后分别将经磷酸酸化后的猪尿和提取后的猪粪溶液经固相萃取小柱富集净化,取净化液进行HPLC分析。HPLC流动相为乙腈(A):柠檬酸/乙酸铵缓冲液(B),梯度洗脱:0~25 min,A 10%~40%;25~30 min,A 40%至10%,荧光检测器的激发波长278 nm,发射波长465 nm。结果表明,ENR和CIP 在尿中的最低检测限(LOD)<0.01 mg/L,在粪中的LOD<0.021 mg/kg,在尿中的最低检测限(LOQ)<0.03 mg/L,在粪中LOQ<0.056 mg/kg,猪尿中的ENR和CIP在0.01~1.0 mg/mL范围内线性关系良好,R²分别为0.9994和0.9992;猪粪中的ENR和CIP在0.02~2.0 mg/mL范围内线性关系良好,R²分别为0.9986和0.9981。ENR在猪粪和猪尿中的回收率分别为79.4%和88.5%,CIP在猪粪和猪尿中的回收率分别为75.8%和89.9%。该方法样品处理简单,检测结果准确可靠,且灵敏度较高,是值得推广的检测方法。

关键词: 恩诺沙星; 环丙沙星; 猪; 排泄物; 高效液相色谱; 检测

Abstract: A HPLC-FLD method was developed for determination of enrofloxacin (ENR) and ciprofloxacin (CIP) levels in feces and urine of pig.The pig feces was ultrasonic extracted by acetonitrile-ammonia, then added trichloroacetic acid to make the extraction acidification.The pig urine was acidulated by phosphoric acid and the extraction of feces solution were enriched and purified by solid phase extraction small column, took purification liquid for HPLC analysis.Conditions of HPLC mobile phase was acetonitrile (A):citric acid/ammonium acetate buffer (B), the procedure of gradient elution was 0 to 25 min, A:10% to 40%;25 to 30 min, A:40% to 10%.The detector of fluorescence excitation wavelength was 278 nm, emission wavelength was 465 nm, chromatographic data were measured and recorded.The results showed that the LOD of ENR and CIP were lower than 0.01 mg/L in urine and 0.021 mg/kg in feces, the LOQ of ENR and CIP were lower than 0.03 mg/L in urine and 0.056 mg/kg in feces.ENR and CIP in the concentration of 0.01 to 1.0 mg/mL levels range had good linear relationship, R² were 0.9994 and 0.9992 in pig urine, respectively;ENR and CIP in the concentration of 0.02 to 2.0 mg/mL levels range had good linear relationship, R² were 0.9986 and 0.9981 in pig feces, respectively.The recovery ratio of ENR were 79.4% and 88.5%, and the recovery ratio of CIP were 75.8% and 89.9% in pig feces and urine.After get on validation, the method was easy in sample processing and testing, the results were accurate, reliable and high sensitivity, which was a worth promoting detection method.

Key words: enrofloxacin; ciprofloxacin; pigs; excreta; HPLC; detection

中图分类号:

S823.9⁺1

徐刚, 王周芹, 宋禾, 房春林, 饶勇. 猪排泄物中恩诺沙星和环丙沙星含量的HPLC检测方法[J]. , 2016, 43(4): 1058-1065.

XU Gang, WANG Zhou-qin, SONG He, FANG Chun-lin, RAO Yong. Method for Determination of Enrofloxacin and Ciprofloxacin in Pig Excreta by HPLC[J]. , 2016, 43(4): 1058-1065.

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