[关键词]
[摘要]
目的 建立一测多评法(QAMS)测定藏茴香中6种咖啡酰奎宁酸类成分含量,并验证该方法在藏茴香质量评价中应用的可行性与适用性。方法 取藏茴香粉末(过三号筛)0.5 g,精密加入70%甲醇 20 mL ,超声处理(250 W、频率 53 kHz)30 min,制备供试品溶液;采用Phenomenex GeminiR C18(250 mm×4.6 mm,5 μm)色谱柱,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,检测波长为 330 nm,体积流量 1.0 mL·min-1,柱温 30 ℃,进行专属性、供试品提取条件、检测波长选择、色谱条件、线性关系、精密度、重复性、稳定性、加样回收率方法学考察,建立异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸 B、异绿原酸 C成分含量检测的 HPLC法;以异绿原酸 A为内参成分,分别计算新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C 5种成分的相对校正因子,分别采用3种不同色谱仪和3种色谱柱进行相对校正因子、相对保留时间耐用性考察,对藏茴香样品同时采用外标法与 QAMS 测定 6 种成分的质量分数,比较 2 种测定方法结果的差异。结果 建立的6种成分的HPLC检测方法的专属性、供试品提取条件、检测波长选择、色谱条件、线性关系、精密度、重复性、稳定性、加样回收率均符合要求;新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C的相对校正因子平均值分别是 1.362、1.257、1.335、1.470、1.134,3种不同色谱仪和 3种色谱柱对相对校正因子、相对保留时间均无明显影响;QAMS与外标法2种方法测定3批藏茴香样品中6种成分得到的结果之间无显著差异。结论 建立的QAMS简便、准确、可靠,可用于藏茴香中6种咖啡酰奎宁酸类成分——异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C的定量分析。
[Key word]
[Abstract]
Objective To establish a method for the content determination of six caffeoylquinic acids components in Carum carvi L. through quantitative analysis of multi-components with single marker (QAMS). To verify the feasibility and applicability of the method in its quality control. Methods Take 0.5 g of C. carvi powder (through the third sieve), accurately add 20 mL of 70% methanol, and conduct ultrasonic treatment (250 W, frequency 53 kHz) for 30 min to prepare the test solution. Phenomenex GeminiR C 18 column (250 mm × 4.6 mm, 5 μm) was used, acetonitrile-0.1% formic acid aqueous solution was used as mobile phase, gradient elution, flow rate was 1.0 mL·min-1, column temperature was 30 ℃, and detection wavelength was 330 nm. Conduct methodological studies on specificity, test article extraction conditions, detection wavelength selection, chromatographic conditions, linear relationship, precision, repeatability, stability, and sample addition recovery. With isochlorogenic acid A as the internal reference component, relative correction factors of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C was established. Three different chromatographs and three chromatographic columns were used to investigate the relative correction factor and relative retention time durability. The external standard method and QAMS were used to simultaneously determine the mass fractions of six components in C. carvi samples, and the differences between the two determination results were compared. Results The specificity, extraction conditions, detection wavelength selection, chromatographic conditions, linear relationship, precision, repeatability, stability, and sample recovery of the established HPLC detection methods for the six components meet the requirements. The relative positive factors of isochlorogenic acid A and neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C were 1.362, 1.257, 1.335, 1.470, and 1.134, respectively. Three different chromatographs and three chromatographic columns had no significant impact on the relative correction factor and relative retention time. There was no significant difference between the calculated values of six components in three batches of samples by the QAMS and the measured values of the external standard method. Conclusion The method established in the study is simple and stable, and can be used for the synchronous quality control of six components (isochlorogenic acid A, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C) in C. carvi.
[中图分类号]
R291.4;927.2
[基金项目]
新疆维吾尔自治区自然科学基金资助项目(2022D01E94);新疆维吾尔自治区天山英才培养计划资助项目(2022TSYCCX0023)