HPLC拆分并测定草豆蔻中(±)-山姜素和(±)-乔松素对映体含量

刘永静; 陈晓芬; 黄冬霆; 王晓颖

doi:10.13748/j.cnki.issn1007-7693.20240945

HPLC拆分并测定草豆蔻中(±)-山姜素和(±)-乔松素对映体含量

Enantioseparation and Content Determination of (±)-Alpinetin and (±)-Pinocembrin in Alpiniae Katsumadai Semen by HPLC

摘要

摘要:
目的建立草豆蔻药材中(±)-山姜素和(±)-乔松素同时手性和非手性分离分析方法，并将所建立的方法应用于草豆蔻药材和不同炒制时间的炮制品中2个成分的对映体的含量测定。
方法采用手性柱-HPLC对山姜素和乔松素进行对映体拆分研究，优化手性柱种类、流动相比例、流速和柱温的色谱条件，并在最佳分离条件下测定不同批次草豆蔻药材和不同炒制时间草豆蔻药材中2个成分对映体的含量。
结果确定了山姜素和乔松素对映体的手性和非手性分离条件为PHENOMENEX Lux i-Amylose-3(4.6 mm×250 mm，5 μm)色谱柱，流动相为乙腈-水(62∶38)，流速为0.4 mL·min⁻¹，柱温为25 ℃，检测波长为290 nm。在最佳分离条件下，山姜素2个对映体和乔松素2个对映体的分离度均>1.5，并且草豆蔻中其他成分并不干扰2个成分对映体的含量测定。所测定的3个批次的草豆蔻药材中(+)-乔松素的含量为1.66~3.06 mg·g⁻¹，而(−)-乔松素的含量为2.27~3.66 mg·g⁻¹，2个对映体的EF值为0.41~0.46；山姜素2个对映体的含量分别为3.00~5.46和3.29~5.63 mg·g⁻¹，2个对映体的EF值为0.48~0.49。而随着炒制时间的增加，山姜素2个对映体的含量均呈现先升高再下降的趋势，两者均是在炒制10 min时呈现含量最大值；乔松素对映体1在未炮制的草豆蒄中含量最大，对映体2则在是炒制10 min 时呈现含量最大值；但是炒制对于2个成分的对映体的EF影响不大。
结论所建立的草豆蔻药材中(±)-山姜素和(±)-乔松素同时手性和非手性分离分析方法准确高、重复性好，可用于基于对映体层面的草豆蔻药材的质量控制和质量评价。

Abstract:
OBJECTIVE To establish the method for the simultaneous chiral and achiral separation of (±)-alpinetin and (±)-pinocembrin in Alpiniae Katsumadai Seme, which was applied to the determination of two enantiomers of each analytes in different batches of Alpiniae Katsumadai Seme and different processing time.
METHODS The enantiomeric separation method was studied by chiral columns-HPLC, as well as the chromatographic conditions including type of chiral column, mobile phase ratio, flow rate and column temperature were optimized. The contents of (±)-alpinetin and (±)-pinocembrin in different batches of Alpiniae Katsumadai Seme and different processing time were determined under the optimal separation conditions.
RESULTS The chiral and achiral separation conditions of the enantiomers of (±)-alpinetin and (±)-pinocembrin were determined as follows: PHENOMENEX Lux i-Amylose-3(4.6 mm×250 mm, 5 μm) as chiral column, mobile phase of acetonitrile-water(62∶38) at a flow rate of 0.4 mL·min⁻¹. The detection wavelength was set at 290 nm at a column temperature of 25 ℃. Under the optimal separation conditions, the resolution of two alpinetin isomers and pinocembrin isomers >1.5, and other components of Alpiniae Katsumadai Seme did not interfere with the content determination of the two analytes. The content of (+)-pinocembrin in three batches of Alpiniae Katsumadai Seme was 1.66–3.06 mg·g⁻¹, whileas the content of (–)-pinocembrin was 2.27–3.66 mg·g⁻¹, with the EF values of the two enantiomers were 0.41–0.46. The contents of the two enantiomers of alpinetin were 3.00–5.46 and 3.29–5.63 mg·g⁻¹, respectively, as well as the EF values of the two enantiomers were 0.48–0.49. With the increase of processing time, the contents of the two enantiomers of alpinetin and pinocembrin increased and reached a maximum value at 10 min. However, a decreasing trend of the contents of alpinetin isomers and pinocembrin isomers was observed, as the treatment time continued to increase. It's worth mentioning that processing of Alpiniae Katsumadai Seme had little effect on EF values of the (±)-alpinetin and (±)-pinocembrin.
CONCLUSION The established method for simultaneous chiral and achiral separation and analysis of (±)-alpinetin and (±)-pinocembrin in Alpiniae Katsumadai Seme is accurate and repeatable, which can be used for quality control and evaluation of Alpiniae Katsumadai Seme based on the enantiomeric layer.

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