摘要
目的建立雄性SD大鼠血浆中司美格鲁肽的定量方法,并应用于司美格鲁肽注射液在大鼠体内单次给药和多次给药的药代动力学研究。方法采用超高效液相色谱-三重四极杆质谱联用技术(UPLC-MS/MS)对大鼠血浆中司美格鲁肽含量进行测定。样品预处理:血浆样品100μL经甲醇-乙腈(3∶7)沉淀后取上清液使用Oasis MAXμElution 96孔板进行固相萃取(SPE);色谱条件:采用Waters CSHTMC18色谱柱(130?,2.1 mm×50 mm,1.7μm),5%乙腈水溶液(含0.1%甲酸)为A相,95%乙腈水溶液(含0.1%甲酸)为B相进行洗脱,流速为0.6 mL·min^(-1),进样体积为10μL;质谱条件:采用电喷雾离子源正离子(ESI+)模式,司美格鲁肽和内标利拉鲁肽的多反应监测(MRM)离子通道为m/z 1029.3→m/z 1238.2和m/z 938.6→m/z 1064.4。最后,在大鼠体内进行司美格鲁肽单次和多次给药的药代动力学研究。结果建立了一种快速且稳健的大鼠血浆中司美格鲁肽含量检测方法,检测时间2.5 min,定量范围为1.00~500 ng·mL^(-1)。方法的选择性、基质效应均符合接受标准。批内和批间准确度在-9.5%~4.1%,批内和批间精密度不超过13.4%。在实验条件下,样品均稳定。大鼠体内单次给药t1/2约7.6 h,多次给药蓄积指数为1.21。结论UPLC-MS/MS能准确定量雄性大鼠血浆中司美格鲁肽含量,结果表明司美格鲁肽在大鼠体内的代谢较快,0.05 mg·kg^(-1)·d^(-1)连续每天给药一次没有明显的药物蓄积。
Objective To establish a method for the quantification of semaglutide in male SD rats plasma,and apply it to the research of pharmacokinetics of semaglutide injection in rats after single and multiple doses.MethodssAn ultra-high performance liquid chromatography tandem mass spectroscopy(UPLC-MS/MS)assay was developed to quantify the semaglutide in rats plasma.Sample pre-treatment:Plasma samples 100μL were precipitated with methanol/acetonitrile(v/v=3/7),then supernatant was transferred to solid phase extraction(SPE)using Oasis MAX 96-wellμElution Plate.Chromatography conditions:Waters CSHTM C18 column(130 A,2.1 mm×50 mm,1.7μm)was selected to separate the component.5%acetonitrile and 95%acetonitrile both containing 0.1%formic acid were mobile phase A and B,respectively.The flow rate was O.6 mL·min^(-1),the injection volume was 10μL.Mass spectrometry conditions:ESI+mode,multiple reaction monitor(MRM)scanning mode.The ion transition of semaglutide and liraglutide(internal standard)were m/z 1029.3→m/z 1238.2 and m/z 938.6→m/z 1064.4,respectively.The method was applied to study the pharmacokinetics of semaglutide in male SD rats after single and multiple doses via subcutaneous injection.Results A fast and robust method was established for the quantification of semaglutide in rat plasma within the linear range of 1.00-500 ng·mL^(-1),while total running time was 2.5 min.The results of method validation for selectivity and matrix effect were within the acceptance criteria.The within-run and between-run accuracies were in the range of-9.5%-4.1%,and within-run and between-run precisions were not more than 13.4%.Semaglutide was demonstrated to be stable under tested conditions.The ti/2 of single administration in rats was about 7.6 h,and the accumulation index of multiple administration was 1.21.Conclusion A fast and robust UPLC-MS/MS method was developed and full validated,which was also useful for the quantification of semaglutide in male rat plasma.These results showed that the rate of metabolism in rats was f
作者
唐琦
劳淑华
王子哲
叶观莲
郭地利
陈倩倩
杨志伟
王燕清
徐朋
TANG Qi;LAO Shu-hua;WANG Zi-zhe;YE Guan-lian;GUO Di-li;CHEN Qian-qian;YANG Zhi-wei;WANG Yan-qing;XU Peng(Livzon Microsphere Technology Co.,Ltd.,Zhuhai 519090,Guangdong Province,China;Liuzon Pharmaceutica Gl roup Inc,Zhuhai 519090,Guangdong Province,China;Livzon New North River Pharmaceutical Co.,Ltd,Qingyuan 511500,Guangdong Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第9期1316-1320,共5页
The Chinese Journal of Clinical Pharmacology
基金
“重大新药创制”国家科技重大专项基金资助项目(2017ZX09201004-016)
广东省“珠江人才计划”引进第六批创新创业团队基金资助项目(2016ZT06Y229)。
