UFLC-MS/MS法同时测定龙胆中7个活性成分的含量

Simultaneous Determination of Seven Active Components in Gentianae Radix Et Rhizoma by UFLC-MS/MS

关键词:  龙胆  环烯醚萜苷  黄酮  超快速液相色谱-串联质谱法

Key words:  Gentianae Radix Et Rhizoma  iridoid glycosides  flavonoids  ultra-fast liquid chromatography-tandem mass spectrometry

摘要:

建立可同时测定龙胆中7个活性成分（马钱苷酸、獐牙菜苦苷、6"-O-β-D-葡萄糖基龙胆苦苷、龙胆苦苷、 獐牙菜苷、异荭草苷和异牡荆素）含量的超快速液相色谱-串联质谱方法。流动相采用0.1%甲酸水溶液(A)-乙腈(B)，色谱柱Shim-Pack XR-ODS(75 mm×3.0 mm, 2.2 μm)，梯度洗脱；采用三重四极杆串联质谱仪，电喷雾电离源，正离子模式监测。上述7个成分线性浓度范围分别为2.5-100 μg?mL-1（r = 0.9996）、1-40 μg?mL-1（r = 0.9995）、1-40 μg?mL-1（r = 0.9995）、5-200 μg?mL-1（r = 0.9996）、0.1-4 μg?mL-1（r = 0.9995）、0.05-2 μg?mL-1（r = 0.9998）和0.01-0.4 μg?mL-1（r = 0.9995）。6批龙胆样品中上述7个成分的含量分别为0.894%~1.072%、0.260%~0.285%、0.223%~0.268%、3.88%~4.05%、0.00726%~0.00898%、0.000840%~0.00195%和0.000779%~0.00104%。本方法灵敏、稳定，可用于龙胆药材的质量控制。

Abstract:

To establish an ultra-fast liquid chromatography with tandem mass spectrometry (UFLC-MS/MS) method for simultaneous determination of seven active components (loganic acid, swertiamarin, 6"-O-β-D-glucosylgentiopicroside, gentiopicroside, swertioside, isoorientin and isovitexin) in Gentianae Radix Et Rhizoma. Chromatographic separation was performed on Shim-Pack XR-ODS column (75 mm×3.0 mm, 2.2 μm). The mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B). The seven analytes were determined with gradient elution. Detection was performed on 3200 QTRAP mass spectrometry equipped with electrospray ionization source in positive ionization mode. Quantification was performed using multiple reactions monitoring (MRM) mode. The linear ranges of the seven analytes were 2.5-100 μg?mL-1 (r = 0.9996), 1-40 μg?mL-1(r = 0.9995), 1-40 μg?mL-1(r = 0.9995), 5-200 μg?mL-1(r = 0.9996), 0.1-4 μg?mL-1(r = 0.9995), 0.05-2 μg?mL-1(r = 0.9998) and 0.01-0.4 μg?mL-1(r = 0.9995), respectively. The content ranges of seven components in six batches were 0.894%~1.072%, 0.260%~0.285%, 0.223%~0.268%, 3.88%~4.05%, 0.00726%~0.00898%, 0.000840%~0.00195% and 0.000779%~0.00104%, respectively. The established method is sensitive and stable, and can provide a research method for the quality control of Gentianae Radix Et Rhizoma.