活血消痛酒质量标准及制备工艺研究

The Study on Quality Standard and Preparation Technology of Huoxue Xiaotong Liquor

关键词:  活血消痛酒  薄层色谱法  高效液相色谱法  中心组合设计  质量标准  工艺优化

Key words:  Huoxue Xiaotong liquor  Thin layer chromatography  High Performance Liquid Chromatography  Central composite design.  Quality standard  technology optimizatio

摘要:

目的：建立活血消痛酒质量标准并对其制备工艺进行优化。方法：采用薄层色谱法(TLC)对活血消痛酒的君药两面 针、三七和臣药血竭鉴别；采用高效液相色谱法(HPLC)测定三七苷R1、人参皂苷Rg1、人参皂苷Rb1 的含量。 色谱柱为Waters X Bridge Shield RPC18(4.6 ×250 mm, 5 μm)，流动相：水(A) - 乙腈(B)，梯度洗脱(0~ 15 min，20%~23%B;15~25 min，23%B；25~ 35 min，23%~40%B；35~36 min，40%~90%B)；流速为 1 mL/min；柱温 30 ℃；检测波长为203 nm；进样量为 10 μL；采用中心组 合设计(CCD)响应面法对活血消痛酒工艺优化。结果：两面针、三七、血竭薄层色谱特征斑点清晰，分离度良好，阴性对照无 干扰；三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1 线性范围内良好(r2>0.999), 阴性对照无干扰、精密度、稳定性、重复性、加样回收 率结果良好，10批活血消痛酒的三种皂苷含量范围为0.4225~0.5982 mg/mL；采用CCD法结合实际情况得出最优工艺：含醇量 为42%、物料比为 16.7 mg/mL、浸泡时间为 7d 。结论：该研究建立的方法操作简便、准确可靠，可用于活血消痛酒的质量标准 研究；本研究通过中心组合设计法，科学、全面准确地找到了活血消痛酒的最优工艺。

Abstract:

Objective: To establish quality standard and optimize the preparation technology of Huoxue Xiaotong liquor. Methods: Thin layer chromatography (TLC)was used to identify sovereign drug including Zanthoxylum nitidum(Roxb.) DC and Panax notoginseng and identify ministerdrug: Resina Draconis; High Performance Liquid Chromatography (HPLC) was used to determine the content of notoginsenoside R1 , ginsenoside Rg1 and ginsenoside Rb1 . Chromatographic column is Waters X Bridge Shield RPC18(4.6 ×250 mm, 5 μm) colum. Mobile phase consisting of water (A) -acetonitrile (B) and gradient elution (0~ 15 min，20%~23%B; 15~25 min，23%B；25~35 min，23%~40%B；35~36 min，40%~90%B). The flow rate was 1 mL/min, column temperature was 30 ℃ , detection wavelength was 203 nm, the injection volume was 10 μL; Central composite design (CCD) -response surface method was used to optimize the preparation technology of Huoxue Xiaotong liquor. Results: The characteristic spots of Zanthoxylum nitidum(Roxb.) DC、Resina Draconis and Panax notoginseng were clear and well separated, and there was no interference from the negative control. The notoginsenoside R1 , ginsenoside Rg1 and ginsenoside Rb1 had a good linear relationship within their respective concentration ranges(r2>0.999). There was no interference from the negative control, and the results of precision, stability, repeatability and recovery rate were good. The total content of three saponins in 10 batches of samples ranged from 0.4225~0.5982 mg/mL; The CCD method was used along with actual situation to obtain the optimal process as follows: alcohol content was 42%, material ratio was 16.7 mg/mL, soaking time was 7 days. Conclusion: The method established in this study is simple, accurate and reliable, and can be used for the study of quality standard of Huoxue Xiaotong liquor. At the same time, the optimal technology of Huoxue Xiaotong liquor was found scientifically, comprehensively and accurately by the method of central composite design.