液相色谱-质谱联用法研究青蒿素在鸡体内的代谢规律

为探究青蒿素（ART）在鸡体内的代谢规律，本研究建立了液相色谱串联三重四级杆质谱检测鸡血浆中ART的方法：即以电喷雾电离源（ESI）作为离子源，以蒿甲醚（ARM）做内标（IS），以ART和ARM的[M+Na]⁺准离子峰做二级质谱扫描，选择m/z 305.2 → 151.3（ART）、m/z 321.0 → 163.1（ARM）两对离子以多监测反应方式（MRM）进行定量分析。结果表明，ART在10~1 000 ng/mL范围内线性关系良好（R²=0.9997），最低检测限为10 ng/mL，回收率在90%~105%之间，日间、日内精密度（RSD）均小于15%。选取30日龄蛋鸡，按照3个剂量（15、40、100 mg/kg）以及口服的次数（1、11次）分成6个组，每组6只，各组分别灌胃给药。给药1次的各组前10次先口服溶剂，第11次按剂量灌服ART，给药11次的各组按剂量换算给药。每组在最后1次给药结束后10、30 min及1、1.5、2、2.5、3、4、6、8 h分别采集300 μL血液加入抗凝管中待测。经检测发现，单一剂量（15、40或100 mg/kg）口服给药后ART在鸡血浆中的达峰时间分别为1、1.5、1.5 h；但上述剂量连续多次给药后检测发现药物代谢明显加快，并有剂量依赖性。综合试验结果，本试验建立的ART检测方法操作简单，灵敏，重现性好，可用于鸡血浆中ART的检测；ART在禽类体内存在明显的自身诱导代谢现象，且剂量越高，其代谢速度越快。

Study on the Metabolism of Artemisinin in Chickens by LC-MS/MS

In order to study the metabolism of artemisinin (ART) in chickens,the LC-MS/MS method was established for the determination of ART in chicken plasm.The electrospray ionization (ESI) source was used as the ion source,and artemether (ARM) was the internal standard(IS).The[M+Na]⁺ quasi ion peaks of ART and ARM were chosed for secondary mass spectrometry.Two pairs of ions,m/z 305.2→151.3 (ART) and m/z 321.0→163.1 (ARM),were selected for quantitative analysis by multi-monitoring reaction (MRM).It was concluded that there was a good linear relationship in the range of 10-1 000 ng/mL (R²=0.9997).In this method,the minimum quantitative limit was 10 ng/mL and the recovery rate was between 90% and 105%.Meanwhile,the intra-day and inter-day precision (RSD) were less than 15%.Thirty-day-old chickens were divided into six groups randomly according to the three doses (15,40 and 100 mg/kg) and the number of oral times (one or eleven).The one-time groups were administered ART at the 11th time,and the solvent was orally administered 10 times before.Eleven-time groups were fed ART for 11 times.After the last oral medication,300 μL blood of each chicken was collected in anticoagulant tube at 10 min,30 min,1 h,1.5 h,2 h,2.5 h,3 h,4 h,6 h and 8 h.The results showed that the concentration of ART in the blood of the chicken which were administered for one time (15,40 and 100 mg/kg) could reach the C_max at 1 or 1.5 h.And the metabolism of ART in chickens could be accelerated by orally administered for more times and has a dose-dependent manner.In conclusion,the method established in this study was simple,sensitive and reproducible for the determination of ART in chicken plasma.There was a self-induced metabolism of ART in chickens,meanwhile,higher the dose,faster the metabolism.