双标线性校正法用于板蓝根的多成分定性分析<sup>*</sup>

李高岩, 杨一荻, 袁鹤飞, 曲范娜, 李慧勇, 孙磊, 马双成, 笔雪艳

doi:10.16155/j.0254-1793.2024-0222

药物分析杂志 >

2024 , Vol. 44 >Issue 9: 1560 - 1566

DOI: https://doi.org/10.16155/j.0254-1793.2024-0222

标准研讨

双标线性校正法用于板蓝根的多成分定性分析^*

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1.黑龙江中医药大学,哈尔滨 150040;
2.黑龙江省药品检验研究院国家药品监督管理局中药质量研究与评价重点实验室黑龙江省中药质量研究与评价重点实验室,哈尔滨 150088;
3.哈尔滨商业大学,哈尔滨 150028;
4.中国食品药品检定研究院,北京 100050

第一作者 Tel:18032432063;E-mail:544642683@qq.com

^**Tel:13836141280;E-mail:65554629@qq.com

收稿日期: 2024-04-03

网络出版日期: 2024-10-17

基金资助

^*黑龙江省中医药科研项目(ZHYCYC2022-004)

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Linear calibration with two reference substances for multi-component qualitative analysis of Radix Isatidis^*

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1. Heilongjiang University of Traditional Chinese Medicine, Harbin 150040, China;
2. Key Labratory of Quality Research and Evaluation of Traditional Chinese Medicines, State Medical Products Administration, Heilongjiang Provincial Key Laboratory of Quality Research and Evaluation of Traditional Chinese Medicines, Heilongjiang Institute for Drug Control, Harbin 150088, China;
3. Harbin University of Commerce, Harbin 150028, China;
4. National Institutes for Food and Drug Control, Beijing 100050, China

Received date: 2024-04-03

Online published: 2024-10-17

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摘要

目的: 建立同时检测板蓝根中6个成分(尿苷、腺嘌呤、鸟苷、(R,S)-告依春、腺苷、直铁线莲宁B)的HPLC方法,考察双标线性校正(LCTRS)法对于板蓝根多成分定性分析的可行性。方法: 采用HPLC法,以甲醇为流动相A,水为流动相B,梯度洗脱(0~3 min,3%A; 3~18 min, 3%A→14%A; 18~25 min, 14%A→26%A; 25~34 min, 26%A; 34~40 min, 26%A→46%A; 40~60 min, 46%A→90%A),流速0.8 mL·min^-1,柱温30 ℃,检测波长为0~32 min,254 nm;32~60 min,230 nm,进样量10 μL;测定板蓝根中6个成分在16根不同品牌和型号的C₁₈色谱柱的实际保留时间,以鸟苷和直铁线莲宁B 2个成分作为双标化合物,采用LCTRS法定位各成分色谱峰,并用3根未知色谱柱进行方法验证。同时以鸟苷为参照物,采用RRT法预测其他5个成分的保留时间。比较这2种方法的预测准确性和色谱柱符合率。结果: LCTRS法能够较好地对6个指标性成分进行保留时间的预测和定性分析。与RRT法相比较,LCTRS法所预测结果准确性更高,色谱柱普适性更优。结论: LCTRS法用于同时测定板蓝根中多成分可行且准确,操作简便,耐用性好,具有推广价值。

关键词： 板蓝根; 双标线性校正法; 鸟苷; 直铁线莲宁B; 尿苷; 腺嘌呤; (R,S)-告依春; 腺苷

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李高岩, 杨一荻, 袁鹤飞, 曲范娜, 李慧勇, 孙磊, 马双成, 笔雪艳 . 双标线性校正法用于板蓝根的多成分定性分析^*[J]. 药物分析杂志, 2024 , 44(9) : 1560 -1566 . DOI: 10.16155/j.0254-1793.2024-0222

Abstract

Objective: To establish an HPLC method for the simultaneous determination of six components (uridine, adenine, adenosine (R,S)-goitrine, guanosine,clemastanin B) in Radix Isatidis, and to investigate the linear calibration with two reference substances (LCTRS) method for the qualitative analysis of multiple components in Radix Isatidis. Methods: HPLC method was used, with methanol as mobile phase A and water as mobile phase B. Gradient elution (0-3 min, 3%A; 3-18 min, 3%A→14%A; 18-25 min, 14%A→26%A; 25-34 min, 26%A; 34-40 min, 26%A→46%A; 40-60 min, 46%A→90%A) was performed at a flow rate of 0.8 mL·min^-1. The column temperature was 30 ℃, the detection wavelengths were 254 nm (0-32 min) and 230 nm (32-60 min). The injection volume was 10 μL. The actual retention time of 6 components in Radix Isatidis was determined on 13 C₁₈ chromatographic columns of different brands and models. Guanosine and clemastanin B were used as double reference compounds, and LCTRS method was used to locate the chromatographic peak of each component. Three unknown chromatographic columns were used for method validation. Using guanosine as a reference substance, the relative retention time method was used to predict the retention time of the other 5 components. The predictive accuracy and column coincidence of these two methods were compared. Results: The LCTRS method could effectively predict and qualitatively analyze the retention time of six indicator components. Compared with the relative retention time method, the LCTRS method had higher accuracy in predicting results and better column universality. Conclusion: The LCTRS method for simultaneous determination of multiple components in Radix Isatidis is feasible and accurate, with simple operation and good durability, and has promotional value.

Key words： Radix Isatidis; linear calibration with two reference substances; guanosine; clemastanin B; uridine; adenine; (R,S)-goitrine; adenosine

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