基于双标线性校正技术的白及多组分定性分析&#x0002A;

林昶; 刘苏情; 张静; 杨聪; 钱海兵; 杨长福; 赵君

doi:10.16155/j.0254-1793.2024-0317

药物分析杂志 >

2025 , Vol. 45 >Issue 3: 392 - 399

DOI: https://doi.org/10.16155/j.0254-1793.2024-0317

成分分析

基于双标线性校正技术的白及多组分定性分析^*

林昶 ,
刘苏情 ,
张静 ,
杨聪 ,
钱海兵 ,
杨长福 ,
赵君

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1.贵州中医药大学,贵阳 550000;
2.贵州同德药业股份有限公司,铜仁 554300

第一作者 Tel：15285966608;E-mail：375718640@qq.com

^** Tel：13984350701;E-mail：279753407@qq.com

收稿日期: 2024-05-14

网络出版日期: 2025-08-25

基金资助

*贵州省科技计划项目：黔产白及产地加工一体化加工技术研究与白及多糖智能化提取加工与应用（黔科合支撑[2022]重点023-02）; 贵州省高等学校中药（民族药）药性与效应研究重点实验室资助项目（黔教技〔2023〕18号）; 贵州中医药大学国家与省级科技创新人才团队培育项目（贵中医TD合字〔2023〕3号）

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Qualitative determination of multiple components in Bletillae Rhizoma by linear calibration using two reference substances^*

LIN Chang ,
LIU Su-qing ,
ZHANG Jing ,
YANG Cong ,
QIAN Hai-bing ,
YANG Chang-fu ,
ZHAO Jun

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1. Guizhou University of Traditional Chinese Medicine, Guiyang 550000, China;
2. Guizhou Tongde Pharmaceuticals Co., Ltd, Tongren 554300, China

Received date: 2024-05-14

Online published: 2025-08-25

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摘要

目的: 运用高效液相色谱（HPLC）法对白及中的6个成分进行分析,通过双标线性校正技术,对成分的保留时间进行预测,探讨其在色谱峰定性上的可行性和准确性,并将该方法与相对保留时间法进行比较。方法: 采用高效液相色谱法,以水为流动相A,乙腈为流动相B,梯度洗脱（0~11 min,12%B;11~58 min,12%B→68%B;58~65 min,68%B→12%B）,流速为0.9 mL · min^-1,进样量为5 μL,柱温为30 ℃,检测波长为280 nm。通过测定16根不同C₁₈色谱柱上6个成分的实际保留时间,计算出标准保留时间。以天麻素（峰1）和手参苷Ⅲ（峰4）作为双标成分,运用双标线性校正技术预测其他成分的保留时间,并在另外3个未知C₁₈色谱柱上进行验证。利用RRT法,以白及苷（峰5）为参照化合物,预测其他5个成分的保留时间。结果: 建立了对白及6个成分的定性分析方法,可以准确预测白及中各个成分的保留时间。6个成分的标准保留时间分别为天麻素4.279 min、手参苷Ⅲ 32.145 min、山慈菇素Ⅰ 27.220 min、2-O-葡萄糖基白及苷28.603 min、白及苷33.509 min、山药素Ⅲ 45.504 min。结论: 双标线性校正法可以提高对各组分保留时间的预测能力,从而提高分析结果的准确性和可靠性。

关键词： 白及; 双标线性校正技术; 相对保留时间法; 标准保留时间; 色谱峰定性

本文引用格式

林昶 , 刘苏情 , 张静 , 杨聪 , 钱海兵 , 杨长福 , 赵君 . 基于双标线性校正技术的白及多组分定性分析^*[J]. 药物分析杂志, 2025 , 45(3) : 392 -399 . DOI: 10.16155/j.0254-1793.2024-0317

Abstract

Objective: To analyze six components in Bletillae Rhizoma using high-performance liquid chromatography. Through the linear calibration using two reference substances method, the retention time of the components was predicted, and the feasibility and accuracy of this technique in the qualitative identification of chromatographic peaks were explored. This method was also compared with the relative retention time (RRT) method. Methods: High-performance liquid chromatography was employed using an Ultimate XB-C₁₈ column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of water (A) and acetonitrile (B), with a gradient elution as follows: 0-11 min,12% B; 11-58 min, 12% B to 68% B; 58-65 min, 68% B to 12% B. The flow rate was 0.9 mL · min^-1, the injection volume was 5 μL, the column temperature was maintained at 30 °C, and the detection wavelength was set at 280 nm. By measuring the actual retention time of the six components on 16 different C₁₈ chromatographic columns, the standard retention time was calculated. Gastrodin (Peak 1) and gymnadenin Ⅲ (Peak 4) were used as the double-standard compounds. The linear calibration using two reference substances method was used to predict the retention time of other components, and the prediction was verified on another three unknown C₁₈ chromatographic columns. With the relative retention time (RRT) method, using bletilloside (Peak 5) as the reference compound, the retention times of the other five components were predicted. Results: A qualitative analysis method for the six components in Bletillae Rhizoma was established, which could accurately predict the retention times of each component in Bletillae Rhizoma. The standard retention times of the six components as as follows: gastrodin 4.279 min, gymnadenin Ⅲ 32.145 min, pleione bulbocodioides factor Ⅰ 27.220 min, 2-O-glucosylbletilloside 28.603 min, bletilloside 33.509 min, and batatasin Ⅲ 45.504 min. Conclusion: The linear calibration using two reference substances method can improve the prediction ability of the retention time of each component in Bletillae Rhizoma, thereby enhancing the accuracy and reliability of the analysis results.

Key words： Bletillae Rhizoma; liner calibration with two reference substances; relative retention time; relative retention time; peak localization in chromatography

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