摘要
目的 建立可靠的全血采集及生物分析方法,用于甲磺司特(suplatast tosilate,ST)颗粒的生物等效性研究。方法 冰浴条件下,将全血采集到含有NaF/EDTA抗凝剂的采血管中,分离的血浆分成2份至采血管中,分别加入体积比为0.5%的甲酸。20μL血浆样品经蛋白沉淀处理后上样,以ST-D5为内标。采用正相硅胶分离体系,色谱柱为Agilent Polaris 3Si-A(3.0 mm×100 mm,3μm),流动相为10 mmol·L^(-1)乙酸铵水溶液和甲醇-乙腈溶液(1︰1),流速0.80 mL·min^(-1),以50%有机相进行等度洗脱。在电喷雾电离源正离子模式下检测,ST和ST-D5离子通道(m/z)分别为328.1→266.3,333.1→271.3。结果 定量范围为1~200 ng·mL^(-1),线性关系良好(r=0.999 8);ST与TS-D5提取回收率均在105.0%~117.1%;批内和批间准确度分别为95.67%~108.00%(精密度≤9.10%)和99.10%~105.00%(精密度≤7.66%)。ST在全血、血浆中的稳定性符合要求。结论 所建方法稳定、准确、重复性好,且所需样本体积少,满足ST颗粒生物等效性研究的需求。
OBJECTIVE To establish a reliable method for whole blood collection and bioanalysis method for the bioequivalence study of suplatast tosilate(ST) granules.METHODS Blood was collected into heparin tubes containing NaF/EDTA under ice bath conditions.After processing,the separated plasma was split into 2 aliquots into each heparin tube containing 0.5% formic acid.Plasma sample(20 μL) was pre-treated by protein precipitation,and ST-D5 was used as the internal standard.Chromatographic separation was achieved through a normal phase silica column[Agilent Polaris 3 Si-A column(3.0 mm×100 mm,3 μm)],the mobile phases was 10 mmol·L^(-1) ammonium acetate solution and methanol-acetonitrile(1︰1).The flow rate was 0.80 mL·min^(-1),the organic proportion was 50% with isocratic elution.Then with the mass spectrometer operated in positive electrospray ionization mode using multiple reaction monitoring,ST and ST-D5 were quantitative analyzed by detecting the ion transitions of m/z 328.1→266.3,333.1→271.3.RESULTS The method showed linearity over the concentration range of 1-200 ng·mL^(-1) was good(r=0.999 8).The extraction recoveries of ST and ST-D5 were between 105.0%-117.1%.Intra-and inter-day accuracy were in the ranges of 95.67%-108.00%(precision≤9.10%) and 99.10%-105.00%(precision≤7.66%),respectively.Stability of ST in whole blood and plasma were acceptable.CONCLUSION The established method is stable,accurate,and reproducible for ST with small sample volume,and meets the requirements of bioequivalence studies of ST granules.
作者
杨再香
谭何新
严紫薇
张宗平
肖玉梅
YANG Zaixiang;TAN Hexin;YAN Ziwei;ZHANG Zongping;XIAO Yumei(Chongqing Liujiang Pharma Tech Co.,Ltd.,Chongqing 401338,China;Zhi’en Biotechnology Co.,Ltd.,Chongqing 400039,China)
出处
《中国现代应用药学》
CAS
CSCD
北大核心
2022年第9期1197-1202,共6页
Chinese Journal of Modern Applied Pharmacy
作者简介
杨再香,女,硕士,E-mail:124394918@qq.com;通信作者:张宗平,男,博士,E-mail:zhangzongping@cqljpharma.com。
