Institute of Chinese Medical Sciences, University of Macau, Taipa, Macao SAR, P. R. China.
J Sep Sci. 2011 Oct;34(19):2576-85. doi: 10.1002/jssc.201100295. Epub 2011 Aug 29.
A high-performance liquid chromatography (HPLC) coupled with triple quadrupole mass spectrometry (MS/MS) method was developed for rapid determination of 13 isoflavones in Radix puerariae. A novel shell-type column, namely Kinetex core-shell C(18) column (50 mm×2.1 mm id, 2.6 μm), and gradient elution were used during the analysis. The chromatographic peaks of 13 investigated compounds were identified by comparing their retention time and MS data with the related reference compounds. Multiple-reaction monitoring (MRM) was employed for the quantitative analysis with negative ionization mode. All calibration curves showed good linearity (r(2)>0.9990) within test ranges. The LOD and LOQ were lower than 0.017 and 0.873 μg/mL on column, respectively. The intra- and inter-day precisions for 13 analytes were <1.17 and 2.17%, respectively, and the recoveries were 93.1-104.4%. The validated method was applied for quantitative analysis of 13 isoflavones in 7 species of Radix puerariae. The result demonstrated that HPLC-MS/MS system with Kinetex column could be a promising analytical tool for the determination of isoflavones in traditional Chinese medicines, which is helpful for comprehensive evaluation of quality of R. puerariae.
建立了一种高效液相色谱-串联三重四极杆质谱（HPLC-MS/MS）方法，用于快速测定葛根中的 13 种异黄酮。采用新型壳型柱 Kinetex 核壳 C18 柱（50mm×2.1mmid，2.6μm），并在分析过程中采用梯度洗脱。通过比较保留时间和 MS 数据与相关参考化合物，鉴定了 13 种被研究化合物的色谱峰。采用负离子模式下的多反应监测（MRM）进行定量分析。所有校准曲线在测试范围内均具有良好的线性（r2>0.9990）。在柱上，LOD 和 LOQ 分别低于 0.017 和 0.873μg/mL。13 种分析物的日内和日间精密度分别小于 1.17%和 2.17%，回收率为 93.1%-104.4%。该方法已用于 7 种葛根中 13 种异黄酮的定量分析。结果表明，采用 Kinetex 柱的 HPLC-MS/MS 系统可以成为一种很有前途的分析工具，用于测定中药中的异黄酮，这有助于对葛根质量进行全面评价。
