【作者】 袁玉花；

【导师】 刘力； 陈杖榴；

【作者基本信息】 西南农业大学 ， 临床兽医学， 2002， 硕士

【摘要】 氯霉素是第一个人工合成的广谱抗生素，因其效高价廉，在我国畜牧业中得到广泛应用，对畜禽疾病的控制和治疗起到了重要的作用。随着氯霉素的广泛应用及研究的深入，发现其有许多毒、副作用，因此，必须控制氯霉素在动物性食品中的残留，保障消费者健康与安全。为了了解鸡蛋中氯霉素残留情况和建立一种准确、可靠的检测鸡蛋中氯霉素残留的方法以监控鸡蛋中氯霉素残留，本文研究了鸡蛋中氯霉素残留的高效液相色谱检测方法及消除规律。 将蛋白、蛋黄制成匀浆，用乙腈、乙酸乙酯、正己烷萃取及净化后，用流动相或35％的乙腈溶液重新溶解定容。紫外检测波长为280nm，流动相为乙腈：水（20：80，v／v），流速为1ml／min，进样量为50μl，最低检测限为0.002μg/g，用Waters Nova-PakC₁₈（3.9×150mm）色谱柱分离检测。在上述条件下，氯霉素在鸡蛋（蛋白和蛋黄）中的最低检测限为0.002μg／g，标准曲线的线性范围为0.0021μg／g～20μg／g。氯霉素浓度为0.002μg/g、0.5μg/g、20μg/g时蛋白的批间变异系数分别为5.29％、3.43％、2.61％；批内变异系数分别为6.03％、3.52％、3.44％（第1批），6.32％、3.47％、1.14％（第2批），3.90％、3.73％、1.10％（第3批），5.58％、2.46％、3.03％（第4批）；蛋黄的批间变异系数分别为5.58％、3.37％、1.59％；批内变异系数分别为6.53％、3.83％、1.34％（第1批），5.07％、2.78％、1.15％（第2批），5.31％、2.44％、1.10％（第3批），6.14％、3.28％、1.25％（第4批）。将氯霉素以0.0021μg／g、0.5μg／g和20μg／g分别添加到空白蛋白和蛋黄中，测得蛋白中氯霉素的回收率±标准误分别为105.65％+2.41％、94.89％±0.95％、91.65％±1.13％，蛋黄中氯霉素的回收率±标准误分别为104.79％±2.70％、83.15％±1.11％、80.27％±0.41％。结果表明，本文所建立的高效液相色谱方法是一种灵敏、可靠的检测鸡蛋（蛋白和蛋黄）中氯霉素残留的分析方法。 选用20只处于产蛋高峰期的健康海兰蛋鸡，单笼饲养，多剂量内服氯霉素（100mg/kg b．w，连喂12d），于停药后1d、2d、3d、4d…逐只逐日收集鸡蛋，用本研究所建立的方法分别检测蛋白、蛋黄中的氯霉素，直至检测不到氯霉素为止。采用MCPKP药物动力学参数计算程序处理蛋白和蛋黄中药物浓度——时间数据。停药当天，蛋白中的药物含量为3.61μg/g，蛋黄中的药物含量为14.51μg/g；停药5d，蛋白中的药物含量为0.05μg/g，蛋黄中的药物含量为7.17μg/g；停药8d，蛋白中已检测不到氯霉素，蛋黄中的药物含量为0.80μg/g；停药11d，蛋黄中尚有0.041μg/g的氯霉素，而停药12d，蛋黄中就检测不到氯霉素的含量。 蛋鸡多剂量内服氯霉素，连续给药12d，停药后，其在蛋白和蛋黄中的药动学主要参数分别为：氯霉素在蛋白中的AUC为9.96 mg／L·d，消除半衰期为0.75d；氯霉素在蛋黄中的AUC为81.46mg／L·d，在蛋黄中的消除半衰期为1.64d。 试验结果表明，蛋鸡多剂量内服氯霉素后，氯霉素在鸡蛋中的药动学特征是：易于吸收，多分布于蛋黄；氯霉素在蛋黄中的残留水平高，且在蛋黄中的消除较蛋白缓慢。更多还原

【Abstract】 Chloramphenicol(CAP) was a high-effective broad-spectrum antibiotics used in animals. But it has been found that CAP residues in animal tissues and/or animal products had toxic effects to human body. Therefore, it was necessary to study an analytical method for the elimination of monitoring CAP residues in eggs.A high-performance-Hquid-chromatography(HPLC) method for measuring CAP residues in eggs was established. CAP residues were extracted and pured by acetonitrile, ethyl acetate, and n-hexane from the albumen and yolk of eggs, and then redissolved by mobile phase (acetonitrile: water,20: 80,v/v) and 35% acetonitrile solution respectively at Iml/min flow rate and 25 癈. Detection wave length of UV was set at 280nm. Under the above conditions, the detection limit of CAP in eggs was 0.002fig/g. The calibration curve was linear in the range of 0.002~20ng/g. At 0.002ng/g,0.5ng/g and 20ug/g concentration, the CV% between run was 5.29%, 3.43%, 2.61% for albumen and 5.58%, 3.37%, 1.59% for yolk respectively; the within run CV% was 6.03%, 3.52%, 3.44% (first batch), 6.32%, 3.47%, I.l4%(second batch), 3.90%, 3.73%, 1.10%(third batch), 5.58%, 2.46%, 3.03%(fourth batch) for albumen and 6.53%, 3.83%, 1.34% (first batch), 5.07%, 2.78%, 1.15%(second batch), 5.31%, 2.44%, 1.10%(third batch), 6.14%, 3.28%, 1.25%(fourth batch) for yolk respectively.The recoveries of CAP added to the albumen and yolk at levels of 0.002ug/g, 0.5ug/g and 20ug/g were 105.65 2.41%,94.89 0.95%,91.65 1.13% from albumen, and 104.79 2.70%,83.15 + 1.11 %,80.27 0.41% from yolk respectively. The results showed that this method was sensitive and reliable to measuring CAP residues in albumen and yolk of eggs.Depletion of CAP residues in eggs after oral administration was also studied. Twenty healthy Hailan laying birds were given CAP (lOOmg/kg b.w) consecutively for 12 days. After the end of administration, the CAP in albumen and yolk of eggs collected from these laying birds was detected everyday by the HPLC method above. The concentration-time data of CAP in albumen and yolk were analyzed with MCPKP computer program. On 1 day after the last-administration, the concentrations of CAP in albumen and yolk were 3.61ug/g and 14.51ug/g respectively. On 5 days of the last administration, the concentrations of CAP in albumen and yolk were 0.05|ig/g and 7.17ng/g respectively. On 8 days of the last administration, no CAP residues were detected in egg-albumen, but 0.80^g/g concentration of CAP residues in yolk was still detectable. On 12 days of the last administration, the CAP residues in egg yolk were below the detection limit. The results indicated that CAP residues in eggs were eliminated slowly, especially in yolk.The main pharmacokinetic parameters of CAP residues in eggs were as follow: AUC=9.96mg/l.day,Ti/2P=0.75d in albumen; AUC=81.46mg/l.day,T|/2 p=1.64d in yolk.The results suggested that the pharmacokinetic characteristics of CAP in eggs manifested a rapid absorption of CAP with higher concentration and slower elimination in yolk than that in egg-albumen.更多还原