UPLC-MS/MS法测定银杏叶分散片中4种萜内酯的含量

Determination of Four Terpene Lactones in Ginkgo biloba leaves Dispersible Tablets by UPLC-MS/MS

目的建立一种适合同时测定银杏叶分散片提取物中4种萜内酯(白果内酯、银杏内酯A、银杏内酯B、银杏内酯C)含量的超高效液相色谱-串联质谱(UPLC-MS/MS)法。方法以芍药苷作为内标物,采用UPLC-MS/MS法,色谱柱为Eclipse Plus C18(50 mm×2.1 mm,1.8μm),流动相为甲醇-0.1%甲酸水溶液,梯度洗脱,流速为0.4 m L·min^(-1),柱温为40℃。质谱采用负离子模式,多反应监测扫描。结果白果内酯在8.34~66.72μg·m L^(-1)浓度范围内线性良好,r=0.999 8,平均回收率为92.9%(RSD=2.42%);银杏内酯A在4.64~37.12μg·m L^(-1)浓度范围内线性良好,r=0.999 4,平均回收率为93.2%(RSD=2.10%);银杏内酯B在3.375~27.00μg·m L^(-1)浓度范围内线性良好,r=0.999 4,平均回收率为92.0%(RSD=1.31%);银杏内酯C在4.74~37.92μg·m L^(-1)浓度范围内线性良好,r=0.999 8,平均回收率为94.0%(RSD=2.59%)。结论建立的UPLC-MS/MS方法可用于同时测定银杏叶提取物中4种萜内酯成分,简单快捷、灵敏度高且专属性好。

OBJECTIVE To establish an ultra-high performance liquid chromatography-tandem mass spectrometry（ UPLC-MS/MS）method for determination of four terpene lactones（ bilobalide,ginkgolide A,ginkgolide B,ginkgolide C） in Ginkgo biloba leaves dispersible tablets extract. METHODS Paeoniflorin was used as an internal standard（ IS）.Eclipese Plus C18 column（ 50 mm×2.1 mm,1.8 μm） was used with the mobile phase consisting of methanol-0.1% fomic acid water,and a gradient elution was used.The flow rate was 0.4 m L·min（-1） and the column temperature was set at 40 ℃. A triple quadrupole mass spectrometer operating in negative electrospray ionization mode with multiple reaction monitoring was used. RESULTS The linear range of bilobalide was 8.34-66. 72 μg·m L（-1）（ r = 0.999 8） with recovery of 92.9%（ RSD = 2.42%）; that of ginkgolide A was 4. 64-37. 12 μg·m L（-1）（ r = 0.999 4） with recovery of 93.2%（ RSD =2.10%）; that of ginkgolide B was 3.375-27.00 μg·m L（-1）（ r= 0.999 4） with recovery of 92.0%（ RSD= 1.31%）; that of ginkgolide C was 4.74-37.92 μg·m L（-1）（ r = 0.999 8） with recovery of 94.0%（ RSD = 2.59%）. CONCLUSION The established UPLC-MS/MS method is simple,sensitive and effective,which can be used for the determination of the four terpene lactones in Ginkgo biloba leaves tablets extract.