HPLC-MS法测定经阴道给药后人血清克林霉素浓度

HPLC-MS determination of clindamycin in human serum after intravaginal administration of clindamycin phosphate vaginal gel

目的:建立健康志愿者克林霉素磷酸酯阴道凝胶阴道局部给药人血清中克林霉素浓度的HPLC-MS分析方法。方法:以莫沙比利为内标,血清样本经氢氧化钠溶液碱化、乙酸乙酯萃取后,以Hypersil ODS2柱(5μm,4.0mm×200mm)为固定相,以乙腈-醋酸铵溶液(5mmol·L^(-1))(55∶45,v/v)为流动相,流速0.8mL·min^(-1),柱温25℃,进样量10μL,采用SIM质谱检测方式测定克林霉素血浓度。结果:人血清克林霉素最低检测浓度为0.5ng·mL^(-1),在0.5～320ng·mL^(-1)范围内线性关系良好(r=0.9998),平均回收率大于88%,日内、日间RSD均小于9.0%。结论:本法专属、准确、灵敏,可用于克林霉素磷酸酯阴道凝胶阴道局部给药人血清克林霉素浓度的检测。

Objective： To establish an accurate and sensitive HPLC - MS method for the determination of clindamycin serum concentrations in healthy Chinese female volunteers after a single dose and multiple doses of 5.0 g clindamycin phosphate vaginal gel. nethods：Mosapride was used as internal standard. The human serum samples were alkalized with 0. 1 mol· L^ - 1 sodium hydroxide solution, extracted with acetic ether and analysed by HPLC - MS method. The seperation was carried on a Hypersil ODS2 column （5μm,4.0mm×200mm） at 25 ℃, and eluted with acetonitrile - 5 mmol ·L^- 1 ammonium acetate solution （55 ： 45 ） at a flow rate of 0. 8 mL ·min^- 1. The detection was performed with positive ion SIM mode of clindamycin （ [ M ＋ H ] ^＋ , m/z 425 ） and mosapride （ [ M ＋ H ] ^＋ , m/ z 422）. Results ：The detection limit of clindamycin in serum was 0. 5 ng · mL^-1 ,with a good linearity in the range of 0. 5 -320 ng· mL^-1（ r =0. 9998）. The average recovery was more than 88% ,and RSD for inter-day and intra -day were both less than 9.0%. The IS and clindamycin were in good separation. Conclusions：The method is proved to be sensitive, accurate, convenient and suitable for the pharmacokinetic study of clindamycin after intravaginal administration of clindamycin phosphate vaginal vaginal gel in healthy women.