江苏细柱五加果实指纹图谱研究

Study on the HPLC-ELSD Fingerprints of Triterpenoid Saponins in Acanthopanax gracilistylus Fruits

目的:研究建立江苏细柱五加果实中三萜皂苷类成分HPLC-ELSD指纹图谱分析方法,为其质量控制和药材鉴别提供依据。方法:采用HPLC-ELSD色谱法,使用AlltimaTM-C18色谱柱(250mm×4.6mm,5μm),流动相为乙腈-甲醇-0.5%醋酸水,梯度洗脱,流速为1.0mL·min-1,柱温为35℃。ELSD检测器条件:漂移管温度106℃,载气为氮气,流速为2.9L·min-1,进样20μL。结果:在选定的色谱条件下确定14个共有峰,各批次药材均具有上述特征,但特征峰的相对含量分布差异导致色谱概貌存在一定差异。确定了细柱五加果实中2个三萜皂苷类成分,相对保留时间31min为chikusetsusaponinⅣa峰(10号),相对保留时间50min为3β-[(O-β-D-glucopyranu-ronosyl]oxy)-olean-12-ene-28-olcacid峰(14号),不同批次的10批细柱五加果实中HPLC-ELSD指纹图谱特征峰分离度好,谱峰匹配一致。结论:该方法准确、简单、稳定,其色谱指纹图谱可用于江苏细柱五加果实的鉴别和质量控制。

Objective： To establish HPLC-ELSD fingerprints of triterpenoid saponins for quality control and authentication of the crude drug of Acanthopanax graeilistylus fruits. Methods： HPLC-ELSD method was developed. For HPLC, the Alhima C18 column （250 mm× 4. 6 mm, 5 μm） was used. The mobile phase consisted of acetonitrile, methanol and 0. 5 % acetic acid aqueous solution with gradient elution, and the flow rate was 1.0 mL· min^- 1 The column temperature was maintained at 35 ℃ ; For ELSD, the temperature of drift tube was 106 ℃, and flow rate of the carrier gas （nitrogen） was set at 2. 9 L-min-1 The injection volume was 20 μL. Results： Under the present chromatographic conditions, 14 common peaks were selected as the characteristic peaks for fingerprints. All tested samples possessed the above characteristic peaks, but the relative peak areas of these peaks were different in various samples, leading to the diversities of their general chromatographic fingerprints. The specific peaks at relative retention time of 31 min （chikusetsusaponinlVa） and 50 min {3β-（ [ O-β-D-glueopyranuronosyl] oxy） -olean-12-ene-28-ole acid/ were confirmed to be oleanane type triterpenoid saponins. The HPLC-ELSD fingerprints for 10 different batches of samples showed high similarities with respect to the characteristic peaks. Conclusion： The method is accurate, simple and consistent, and can be used for the authentication and quality control of Acanthopanax graeilistylus fruits.