| 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(3.0 mm×100 mm,3.5 μm),检测波长: 除异甘草素外的5种成分采用235 nm检测,异甘草素采用370 nm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+5 mmol/L醋酸铵),B: 0.1%甲酸水溶液(5 mmol/L醋酸铵),梯度洗脱,A相随时间的变化: 25%～35%(0～5 min),35%～50%(5～15 min),50%～85%(15～20 min)；流速0.5 mL/min；柱温25℃；进样量5 μL。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、4.920～98.4 μg/mL范围内线性良好(r>0.999 0),在15 min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。 |
| 关键词: 四逆汤 高压液相色谱法 中药理化鉴定 |
| DOI：10.3724/SP.J.1008.2013.00682 |
| 投稿时间：2013-01-06修订日期：2013-04-25 |
| 基金项目:上海市药物(中药)代谢产物研究重点实验室开放基金 |
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| Simultaneous determination of six marker components in Sini decoction by high performance liquid chromatography |
| LIU Min1,ZHANG Hai2,CAI Ya-mei3,ZHANG Guo-qing2,CHAI Yi-feng3* |
(1. The Institute of Quality Control of Medical Material and Equipment, The Jiont Logistic Department, PLA Nanjing Miliary Area Command, Nanjing 210002, Jiangsu, China
2. Department of Pharmacy, Eastern Hepatobiliary Hospital, Second Military Medical University, Shanghai 200438, China
3. Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
*Corresponding author.) |
| Abstract: |
| 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,3.5 μm); 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% H2O (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 min), 50%-85% (15-20 min); flow speed: 0.5 mL/min; temperature of column: 25℃; injection volume:5 μL. Sini decoction was obtained by using the traditional decoction preparation method. Results Benzoylmesaconine, benzoylaconine, benzoylhyaconine, isoliquiritigenin, glycyrrhizic acid and 6-gingerol were separated at baseline within 15 min, 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. |
| Key words: Sini decoction high-pressure liquid chromatography TCD physic chem identific |
