高效液相色谱-串联质谱法研究克伦特罗在猪毛发中残留及蓄积代谢规律

摘要

Objective To develop a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for detecting the residues of clenbuterol in pig hair and investigate the residue and elimination regularity of clenbuterol.Methods Growing white pigs (the average weight of 50 kg) were fed with the diet that contained clenbuterol for 35 d, withdrawal 32 d. Three hair samples were collected at each time point (d 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 32, and d 35 after the first drug administration). The hairs were dissolved by sodium hydroxide, extracted by tert-butyl alcohol: ethyl acetate (3:7,v:v), after purified with MCX solid phase extraction cartridge, then the residual amount of clenbuterol in these samples was determined by HPLC-MS/MS. Three pigs were killed at each time point (d 0, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 26, 27, 30, and d 32 after the last drug administration), their hair samples were taken and the residues of clenbuterol were determined by LC-MS/MS.Results The residual amount of clenbuterol in the hair samples presented a rising trend while the drug accumulate in the pig (866.75 µg/kg at 35 d). The residual amount of clenbuterol rose at first, and then reduced slowly after withdrawal. The residual amount of clenbuterol was still to 87μg/kg after withdrawal 32 days.Conclusion This is a simple operation, with a good repeatability and high recovery rate, which is suitable to detect the residue of clenbuterol in animal’s hair. This study can help to offer the reference and basis for supervision of edible animal feeding process.%目的：建立一种检测动物毛发中克伦特罗含量的高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)并研究克伦特罗在毛发中的残留及蓄积代谢规律。方法选择成长期白猪(平均体质量50 kg),饲喂克伦特罗日粮,喂药周期为35 d,停药32 d。分别于喂药后的d 2、5、7、9、11、13、15、17、19、21、23、25、27、32、35,每个时间点随机采毛发样品3份, l.0 moL/L氢氧化钠溶液溶解,叔丁醇：乙酸乙酯(3：7,v：v)震荡萃取,过阳离子交换(mixed-mode cationic exchange, MCX)固相萃取柱净化后用LC-MS/MS法检测克伦特罗残留量。于休药后的d 0、3、5、7、9、11、13、15、17、19、21、23、25、26、27、30、32,每个时间点简单随机直抽法处死3只白猪,取毛发样品处理后用HPLC-MS/MS法检测克伦特罗残留量。结果克伦特罗在毛发中的残留浓度随用药增加呈上升趋势(喂药d 35为866.75µg/kg)；克伦特罗在毛发中的残留在休药后浓度先升高后慢慢降低,休药d 32仍能够检出量为87μg/kg。结论该方法操作简单、重复性好、回收率高,适于检测动物毛发中的克伦特罗含量,可为食用动物饲养过程的有效监管提供技术依据。