摘要
目的 建立测定大鼠血浆中汉黄芩素浓度的液相色谱 串联质谱法。方法 血浆样品经液 液萃取后 ,进行色谱分离 ,采用三重四极杆串联质谱检测 ,使用大气压化学电离源 (APCI) ,在正离子条件下选择反应监测 (SRM)方式进行扫描 ,用于定量分析的离子分别为m z 2 84 8→m z 2 6 9 5 (汉黄芩素 )和m z 2 5 4 7→m z 198 5 (葛根黄豆苷元 ,内标 )。结果 线性范围为 0 2 5～ 2 0ng·mL- 1 ,最低定量浓度为 0 2 5ng·mL- 1 。以质控样品 (QC)计算 ,在各浓度水平下 ,此法的批内精密度 (RSD)为 2 2 %～ 13 1% ,批间精密度为 5 9%～ 7 3% ;准确度 (RE)为 - 0 3%～ 1 3%。结论此法灵敏、快速、准确 。
AIM To develop a sensitive and rapid LC/MS/MS method for the determination of wogoni n, an active flavonoid shown to have an inhibitory effect on the proliferation o f a carcinoma cell line, in rat plasma after an oral administration. METHODS Wogonin and daidzein (internal standard) were extracted from plasma directly wit h n hexane diethyl ether (1∶4). After liquid liquid extraction, the anal ytes of interest were separated on a Diamonsil C 18 column. The mobile phas e was acetonitrile water formic acid (80∶20∶1) with a flow rate of 0 8 mL ·min -1 . A Finnigan TSQ (triple stage quadruple) tandem mass spectromete r equipped with an atmospheric pressure chemical ionization (APCI) source was us ed as detector and was operated in positive ion mode. Selected reaction monitori ng (SRM) was used and transitions selected for quantitation were: m/z 284 8 →269 5 for wogonin and m/z 254 7→198 5 for daidzein. The mass spectrome tric conditions were as follows: The temperatures of the vaporizer and heated ca pillary were 450℃ and 250℃, respectively. The corona discharge current was 4 00 μA. Nitrogen was used as the sheath and auxiliary gas, whose settings were 0 6 MPa and 3 mL·min -1 , respectively. Argon was used as the collision gas at a pressure of 1 4 Pa. The collision energy of 35 V was chosen for both wogonin and daidzein. RESULTS The calibration curve was linear over the concentration range of 0 25 to 20 0 ng·mL -1 . The limit of quantitation was 0 25 ng·mL -1 . With in day and between day precision expressed by relative standard deviation (RSD ) was 2 2% to 13 1% and 5 9% to 7 3%, respectively, and the accuracy express ed by RE was -0 3% to 1 3%. CONCLUSION This method proved to be specific, accurate and sensitive enough to be applied t o the pharmacokinetic studies of wogonin in rats after a single dose of 5 mg· kg -1 by oral administration.
出处
《药学学报》
CAS
CSCD
北大核心
2002年第5期362-366,共5页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目 (3 993 0 180 )
