基于HPLC-MS/MS比较不同产地山楂叶中8个成分含量  

Comparison of eight components in hawthorn leaves from different habitats by HPLC-MS/MS

作　　者：潘海峰[1] 杨明宇 徐宝欣 李艳荣[1] 

机构地区：[1]承德医学院河北省中药研究与开发重点实验室,承德067000 [2]沧州医学高等专科学校,沧州061001

出　　处：《药物分析杂志》2017年第12期2173-2179,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：河北省高等学校科学技术研究项目(ZD2015097);河北省高等学校科学研究计划项目(冀教科[2015]7号)

摘　　要：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-C_(18)色谱柱(2.1 mm×100 mm,2.7μm),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 m L·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、0.625~20.0μg·m L^(-1);R^2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 hawthom leaves from different habitats. Meanwhile, the linear ranges of them were 0.719-23.0μg·mL-1 （ R2=0.996 7 ）, 0.688-22.0 μg·mL-1 （ R2=0.998 9 ）, 0.344-11.0μg·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 ）, 2.31-74.0μg·mL-1（ R2=0.998 6 ）, 1.03-33.0 μg·mL-1（R2=0.997 7 ）and 0.625-20.0μg·mL-1（ Rz=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. Condusion： 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 re

关 键 词：山楂叶 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 

分 类 号：R917[医药卫生—药物分析学]