本文旨在建立生物样品中MDMB-4en-PINACA及其代谢标记物的高效液相色谱-三重四极杆串联质谱(HPLC-MS/MS)检测方法,探索MDMB-4en-PINACA及其水解代谢物(M1)和脱烷基代谢物(M2)在大鼠体内降解规律及动态分布规律.将5只SD大鼠分为5组,一组为空白对照组,另外四组灌胃给药MDMB-4en-PINACA后分别置于干净代谢笼中,分别收集1~15 d的尿液;将60只SD大鼠随机分为15组,一组作为空白对照组,其余14组通过灌胃给药MDMB-4en-PINACA,分别在不同时间点(15 min、30min、45 min、1 h、1.5 h、2h、3h、4h、5h、6h、7h、8h、10h、12h)处死,立即取血、尿及组织(心、肝、脾、肺、肾、脑、肌肉).用HPLC-MS/MS检测血液、尿液、各组织中MDMB-4en-PINACA及代谢标记物M1和M2的质量浓度.MDMB-4en-PINACA进入大鼠体内后,迅速分布代谢,各组织和血液中均在15 min内达最高浓度,MDMB-4en-PINACA在体内的分布特点为:脑>脾>血>肺>心脏>肝>肾>肌肉>尿;M1的分布特点为:血>脾>肺>脑>肝>肾>心脏>肌肉>尿;M2的分布特点为:血>肝>心脏>肺>脑>脾>肾>肌肉>尿.在0~12h内MDMB-4en-PINACA及代谢标记物在各组织中呈不同规律的降解.在对1~15 d尿液的监控中,MDMB-4en-PINACA及其代谢标记物仅在少量样本中检出.本文建立的方法操作简单、检出限低、回收率高、重现性好,适用于生物样品中MDMB-4en-PINACA及代谢标记物的检验.经灌胃MDMB-4en-PINACA进入大鼠体内后,监测MDMB-4en-PINACA及代谢标记物在大鼠体内的动态分布及降解规律为体内MDMB-4en-PINACA检验提供科学依据.
To establish a high performance liquid chromatography-triple quadrupole tandem mass spectrometry(HPLC-MS/MS)method for the detection of MDMB-4en-PINACA and its metabolites in biological samples,we studied the degradation and dynamic distribution of MDMB-4en-PINACA,its hydrolytic metabolites(M1),and its dealkylated metabolites(M2)in rats.Five SD rats were divided into five groups,one group was the blank control group,and the other four groups were placed in clean metabolic cages after intragastric administration of MDMB-4en-PINACA.Urine samples were collected from each group for duration of 1 to 15 days respectively.Sixty SD rats were randomly divided into 15 groups;one group was used as the blank control group,and the other 14 groups were administrated with MDMB-4en-PINACA by gavage.They were killed at different time points(15 min,30 min,45 min,1 h,1.5 h,2 h,3 h,4 h,5 h,6 h,7 h,8 h,10 h,and 12 h).Blood,urine,and visceral organs(heart,liver,spleen,lungs,kidneys,brain,and muscles)were collected immediately.The mass concentrations of MDMB-4en-PINACA and metabolites Ml and M2 in the tissues,blood,and urine were detected by HPLC-MS/MS.Following the administration of MDMB-4en-PINACA into the rat's body,it was rapidly distributed and metabolized.Highest concentration was reached in all the tissues and blood samples within 15 min.The distribution characteristics of MDMB-4en-PINACA in the body were as follows:brain>spleen>blood>lung>heart>liver>kidney>muscle>urine.The distribution characteristics of Ml were as follows:blood>spleen>lung>brain>liver>kidney>heart>muscle>urine.The distribution of M2 was as follows:blood>liver>heart>lung>brain>spleen>kidney>muscle>urine.Within 12 hours,MDMB-4en-PINACA and its metabolites were distributed and degraded in different tissues.The contents of MDMB-4en-PINACA and its metabolites were less in the urine monitoring for 1 to 15 days.The established method has the advantages of simple operation,low detection limit,high recovery,and good reproducibility.This method is suitable for the detection of MDMB-4en-PINACA and its metabolites in biological samples.Following the intragastric administration of MDMB-4en-PINACA to rats,its distribution,degradation,and metabolites were monitored to provide a scientific basis for the MDMB-4en-PINACA test in vivo.