高效液相色谱法同时测定四逆汤中6个指标性成分  

Simultaneous determination of six marker components in Sini decoction by high performance liquid chromatography

作　　者：刘敏[1] 张海[2] 蔡亚梅[3] 张国庆[2] 柴逸峰[3] 

机构地区：[1]南京军区联勤部药品仪器检验所,南京210002 [2]第二军医大学东方肝胆外科医院药材科,上海200438 [3]第二军医大学药学院药物分析学教研室,上海200433

出　　处：《第二军医大学学报》2013年第6期682-686,共5页Academic Journal of Second Military Medical University

摘　　要：目的采用高效液相色谱法同时测定四逆汤中苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰乌头次碱、异甘草素、甘草酸和6-姜酚6个成分的含量。方法采用Waters Terra C18色谱柱(3.0mm×100mm,3.5μm),检测波长:除异甘草素外的5种成分采用235nm检测,异甘草素采用370nm的检测波长;流动相为A:95%乙腈和5%水的混合溶液(0.1%甲酸+5mmol/L醋酸铵),B:0.1%甲酸水溶液(5mmol/L醋酸铵),梯度洗脱,A相随时间的变化:25%～35%(0～5min),35%～50%(5～15min),50%～85%(15～20min);流速0.5mL/min;柱温25℃;进样量5μL。四逆汤按照传统煎煮方法提取。结果 6个指标性成分苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰乌头次碱、异甘草素、甘草酸、6-姜酚分别在5.600～112.0、6.560～131.2、6.130～122.6、4.590～91.8、31.00～620.0、4.920～98.4μg/mL范围内线性良好(r>0.999 0),在15min内实现完全分离。方法学考察表明,日内及日间精密度RSD<5%,加样回收率(n=6)分别为101.07%(RSD=1.3%)、98.72%(RSD=1.1%)、101.57%(RSD=1.8%)、101.71%(RSD=3.6%)、102.12%(RSD=2.3%)、99.58%(RSD=3.8%)。结论该方法简便、准确、实用性强,可用于四逆汤中6个指标性成分的含量测定。Objective To simultaneously determine the contents of six marker components, including benzoylmesaconine, benzoylaconine, benzoylhyaconine, isoliquiritigenin, glycyrrhizic acid, and 6-gingerol, in Sini decoction by high performance liquid chromatography （HPLC）. Methods The HPLC condition was as follows： column： Waters Terra C18 （3.0 mm×100 mm,3.5μm）; the detective wavelength was set at 370 nm for isoliquiritigenin and 235 nm for the other five components. Mobile phase： A was 95 % acetonitrile＋ 5 %~ H20 （5 mmol/L ammonium acetate）, B was 0.1% formic acid aqueous solution （5 mmol/L ammonium acetate）, with gradient elution, the gradient of A phase：25-35% （0-5 min）,35%-50% （5-15 rain）, 50%-85% （15 20 min）; flow speed： 0.5 mL/min; temperature of column： 25~C; injection volume：5 μL. Sini decoction was obtained by using the traditional decoction preparation method. Results Benzoylmesaconine, benzoylaconine, benzoylhyaconine, isoliquiritigenin, glycyrrbizic acid and 6-gingerol were separated at baseline within 15 rain, showing good linearity （r〉0. 999 0） between （5. 600-112.0） μg/mL, （6. 560-131.2） μg/mL, （6. 130-122.6） μg/mL, （4. 590 91.8）μg/mL, （31.00-620.0）μg/mL, and （4. 920-98.4）μg/mL, respectively. The results of intra-day and inter-day precisions were at normal range （RSD〈5%）, with the recovery rates （n=6） being 101.07% （RSD=1.3%）, 98.72% （RSD=1. 1%）, 101.57% （RSD=1. 8%）, 101.71% （RSD=3.6%）, 102.12% （RSD=2.3%）, and 99.58% （RSD=3.8%）, respectively. Conclusion The present method is rapid, simple, and accurate and it can be used to determine the above 6 components in Sini decoction.

关 键 词：四逆汤 高压液相色谱法 中药理化鉴定 

分 类 号：R284.1[医药卫生—中药学]