液相色谱-质谱联用测定鼠血浆中的23-羟基白桦酸  

Determination of 23-Hydroxybetulinic Acid in Mouse Plasma by Liquid Chromatogram-Mass Spectrometry

作　　者：杨敏[1,2] 王广基[2] 王素军[2] 李晓天[2] 徐宇平[1] 曹国宪[1] 叶文才[3] 

机构地区：[1]江苏省原子医学研究所,卫生部核医学重点实验室 [2]中国药科大学药代中心,南京210009 [3]中国药科大学天然药化教研室

出　　处：《中国药学杂志》2006年第21期1660-1662,共3页Chinese Pharmaceutical Journal

基　　金：江苏省自然基金资助项目(BK2004025)

摘　　要：目的 建立测定小鼠血浆中230羟基白桦酸的液相色谱0质谱联用法（LC-MS）。方法 取血浆，加入内标物枸橼苦素．经二氯甲烷-水（6：1）提取后，40℃水浴中氮气吹干，残渣用甲醇溶解，进行LC-MS测定。色谱柱为Intersil C18（2.1mm×250mm．3．5μm）；体积分数为70％乙腈为流动相（其中三乙胺为体积分数0．05％）；流速0．2mL·min^-1；柱温30℃；电喷雾离子源（ESI），负离子检测（23-HBA，m／z 471；柠檬苦素，m／z 469）。结果 23-羟基白桦酸的线性范围为10～1000μg·L^-1。（r=0．9998），最低定量限为10μg·L^-1，方法平均回收率〉88％，日内、日间精密度（RSD）均小于8％。小鼠血浆样品-20℃放置至少3周内稳定。结论 本方法专属性强，灵敏度高，线性范围宽，操作简便，适用于药动学研究．OBJECTIVE To develop a sensitive and specific LC-MS method for the determination of 23-hydroxybetulinic acid in mouse plasma.METHODS 23-Hydroxybetulinic acid and the internal standard limonin were extracted by liquid-liquid extraction. The organic layer was blowed by N2 to dryness at 40 ℃ .The residue was reconstituted in methanol. The LC-MS method was performed on an Intersil Cs column （2.1 mm × 250 mm, 3.5μm）. The mobile phase consisted of 70% acetonitrile and 0.05% triethylamine. The flow rate was at 0.2 mL · min^-1.The temperature of column was 30℃ Negative ion electrospray ionization（ESI）was used to form deprotonated molecules at m/z 471of 23-hydroxybetulinic acid and 469 of the internal standard limonin. RESULTS The linear calibration curve was observed in the concentration range of 10 - 1 000μg·L^- 1 （ r = 0.999 8）. The limit of quantitation of 23-hydroxybetulinic acid was 10μg·L^- 1 . The relative recoveries were more than 88% .The inter and intra-day precisions（ RSD）were below 8.0%. 23-Hydroxybetulinic acid was stable in plasma samples at - 20℃ for at least 3 weeks. CONCLUSION The method is proved to be convenient, sensitive, rapid and suitable for pharmacokinetic study.

关 键 词：液相色谱-质谱 23-羟基白桦酸 枸橼苦素 血浆 

分 类 号：R917[医药卫生—药物分析学]