基于HPLC-MS/MS比较不同产地山楂叶中8个成分含量

目的：建立HPLC-MS/MS法同时测定山楂叶中表儿茶素、绿原酸、牡荆素、金丝桃苷、牡荆素鼠李糖苷、牡荆素葡萄糖苷、芦丁、山楂叶苷A 8个成分含量，比较它们在不同产地山楂叶中的含量。方法：采用Poroshell 120 SB-C₁₈色谱柱（2.1 mm×100 mm，2.7 μm），以0.1%甲酸水为流动相A，乙腈为流动相B，梯度洗脱，流速0.15 mL·min^-1，柱温30℃，检测波长320 nm；采用电喷雾离子源（ESI），负离子采集，多反应监测（MRM）模式，对牡荆素葡萄糖苷等8个成分进行指认和含量测定。结果：指认了表儿茶素、绿原酸、牡荆素、金丝桃苷、牡荆素鼠李糖苷、牡荆素葡萄糖苷、芦丁、山楂叶苷A 8个成分，并对其进行含量测定，线性范围分别为0.719~23.0、0.688~22.0、0.344~11.0、0.625~20.0、4.44~142、2.31~74.0、1.03~33.0、0.625~20.0 μg·mL^-1；R²分别为0.996 7、0.998 9、0.999 6、0.993 5、0.997 3、0.998 6、0.997 7、0.995 6。平均加样回收率（n=6）分别为101.7%（RSD=0.89%）、99.41%（RSD=2.6%）、96.20%（RSD=2.4%）、99.56%（RSD=2.7%）、101.3%（RSD=1.5%）、103.8%（RSD=1.8%）、99.82%（RSD=1.6%）、100.2%（RSD=1.5%）。8个成分在33批样品中含量差别较大，表儿茶素、牡荆素、芦丁和山楂叶苷A在有的样品中检测不到，绿原酸、金丝桃苷、牡荆素葡萄糖苷和牡荆素鼠李糖苷在样品中均能检测到。结论：该方法准确可靠，尤其增加了紫外吸收弱、不宜采用HPLC-UV检测的表儿茶素和山楂叶苷A的测定，为不同产地山楂叶质量控制提供依据。

Objective: To establish an HPLC-MS/MS method for simultaneous determination of eight components(epicatechin,chlororenic acid,vitexin,hyperoside,vitexin-rhamnoside,vitexin-glucoside,rutin and shanyenoside A) in hawthorn leaves and compare the contents in hawthorn leaves from different habitats.Methods: The HPLC-MS/MS analysis was conducted on a Poroshell 120 SB-C₁₈ column(2.1 mm×100 mm,2.7 μm) maintained at 30℃.0.1% formic acid(A) and acetonitrile(B) were used as the mobile phase in gradient elution mode.The flow rate was 0.15 mL·min^-1.Electrospray ionization source was used with negative ion scanning and multiple reaction monitoring(MRM) mode for quantification.Results: Eight components(epicatechin,chlororenic acid,vitexin,hyperoside,vitexin-rhamnoside,vitexin-glucoside,rutin and shanyenoside A) were identified and quantified in hawthorn leaves from different habitats.Meanwhile,the linear ranges of them were 0.719-23.0 μg·mL^-1(R²=0.996 7),0.688-22.0 μg·mL^-1 (R²=0.998 9),0.344-11.0 μg·mL^-1(R²=0.999 6),0.625-20.0 μg·mL^-1(R²=0.993 5),4.44-142 μg·mL^-1(R²=0.997 3), 2.31-74.0 μg·mL^-1(R²=0.998 6),1.03-33.0 μg·mL^-1(R²=0.997 7) and 0.625-20.0 μg·mL^-1(R²=0.995 6),respectively. The average recovery rates of them were 101.7%(RSD=0.89%),99.41%(RSD=2.6%),96.20%(RSD=2.4%),99.56%(RSD=2.7%),101.3%(RSD=1.5%),103.8%(RSD=1.8%),99.82%(RSD=1.6%) and 100.2%(RSD=1.5%),respectively.Contents of 8 components had significant differences in 33 batches.Epicatechin,vitexin,rutin and shanyenoside A were not detected in some samples.However,chlorogenic acid,hyperoside,vitexin-glucoside and vitexin-rhamnoside were detected in all samples.Conclusion: The developed HPLC-MS/MS method was accurate and reliable,and it especially proposed the determination method for epicatechin and shanyenoside A with poor UV absorption,thus providing reference for the quality control of hawthorn leaves from different habitats.