Tasly R&D Institute, Tianjin Tasly Group Co., Ltd., Tianjin 300402, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Sep 1;904:51-8. doi: 10.1016/j.jchromb.2012.07.010. Epub 2012 Jul 20.
A rapid, sensitive and reliable liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous determination of five active flavonoids (wogonin, chrysin, oroxylin A, naringenin, hesperetin) and four major alkaloids (berberine, coptisine, jatrorrhizine, palmatine) from Tang-Min-Ling-Pill in rat plasma. Plasma samples (100 μL) were spiked with internal standards daidzein (for flavonoids) and tetrahydropalmatine (for alkaloids), acidified with HCl and extracted by liquid-liquid extraction with acetone. Chromatographic separation was performed on a Zorbax SB-C(18) column with the mobile phase of water (containing 0.1% of formic acid)-acetonitrile (30:70, v/v) at a flow rate of 0.3 mL/min in a run time of 7.0 min. Detection was performed by multiple reaction monitoring mode using electrospray ionization in the positive ion mode. All analytes showed good linearity over the investigated concentration range (r>0.9900). The validated lower limit of quantification was 1.01 ng/mL for wogonin and oroxylin A, 0.238 ng/mL for chrysin, 1.01 ng/mL for naringenin, 0.998 ng/mL for hesperetin, 0.0505 ng/mL for berberine, 0.0996 ng/mL for coptisine, 0.0501 ng/mL for jatrorrhizine, 0.0889 ng/mL for palmatine, respectively. Intra- and inter-day precision (RSD%) was less than 15% and accuracy (RE%) ranged from -7.5% to 4.5%. The validated method was successfully applied to investigate the pharmacokinetics of the major flavonoids and alkaloids of Tang-Min-Ling-Pill after oral administration to rats.
建立并验证了一种快速、灵敏、可靠的液相色谱-串联质谱（LC-MS/MS）法，用于同时测定大鼠血浆中 5 种活性黄酮类化合物（汉黄芩素、白杨素、盐酸小檗碱、柚皮苷、橙皮素）和 4 种主要生物碱（盐酸小檗碱、盐酸巴马汀、盐酸药根碱、盐酸黄连碱）。血浆样品（100 μL）用内标黄豆黄苷（用于黄酮类化合物）和延胡索乙素（用于生物碱）进行预处理，用 HCl 酸化后，用丙酮进行液-液萃取。色谱分离在 Zorbax SB-C18 柱上进行，以水（含 0.1%甲酸）-乙腈（30:70，v/v）为流动相，流速为 0.3 mL/min，运行时间为 7.0 min。采用电喷雾正离子模式进行多反应监测模式检测。所有分析物在研究浓度范围内均表现出良好的线性（r>0.9900）。汉黄芩素和盐酸小檗碱的定量下限为 1.01 ng/mL，白杨素的定量下限为 0.238 ng/mL，柚皮苷的定量下限为 1.01 ng/mL，橙皮素的定量下限为 0.998 ng/mL，盐酸小檗碱的定量下限为 0.0505 ng/mL，盐酸巴马汀的定量下限为 0.0996 ng/mL，盐酸药根碱的定量下限为 0.0501 ng/mL，盐酸黄连碱的定量下限为 0.0889 ng/mL。日内和日间精密度（RSD%）均小于 15%，准确度（RE%）在-7.5%至 4.5%之间。该方法已成功应用于大鼠口服给药后唐敏灵丸主要黄酮类和生物碱的药代动力学研究。
