| 基于发酵过程的淡豆豉HPLC-ELSD特征图谱建立及指标性成分含量测定 |
| 投稿时间：2023-11-23 点此下载全文
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| 引用本文：付予劼,张琳,白莹,袁继巧,王满元.基于发酵过程的淡豆豉HPLC-ELSD特征图谱建立及指标性成分含量测定[J].中国现代中药,2024,26(11):1892-1899 |
| DOI：10.13313/j.issn.1673-4890.20231123007 |
| 摘要点击次数: 286 |
| 全文下载次数: 0 |
| 作者中文名 | 作者英文名 | 单位中文名 | 单位英文名 | E-Mail |
| 付予劼 |
FU Yu-jie |
首都医科大学 中医药学院，北京 100069 |
School of Traditional Chinese Medicine, Capital Medical University, Beijing100069, China |
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| 张琳 |
ZHANG Lin |
首都医科大学 中医药学院，北京 100069 |
School of Traditional Chinese Medicine, Capital Medical University, Beijing100069, China |
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| 白莹 |
BAI Ying |
首都医科大学 中医药学院，北京 100069 |
School of Traditional Chinese Medicine, Capital Medical University, Beijing100069, China |
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| 袁继巧 |
YUAN Ji-qiao |
首都医科大学 中医药学院，北京 100069 |
School of Traditional Chinese Medicine, Capital Medical University, Beijing100069, China |
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| 王满元* |
WANG Man-yuan |
首都医科大学 中医药学院，北京 100069 |
School of Traditional Chinese Medicine, Capital Medical University, Beijing100069, China |
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| 基金项目:国家自然科学基金项目（82274097） |
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| 中文摘要:目的 研究发酵过程中淡豆豉样品化学成分变化的特点，建立可用于淡豆豉质量评价的高效液相色谱-蒸发光散射检测法（HPLC-ELSD）特征图谱，并对淡豆豉中大豆异黄酮类和大豆皂苷类生物活性成分进行含量评价。方法 采用Agilent ZORBAX SB C18色谱柱（150 mm×4.6 mm，3.5 μm），流动相A为0.1%乙酸水溶液，流动相B为0.1%乙酸乙腈溶液，梯度洗脱，流速为1.0 mL·min–1，柱温为30 ℃，蒸发光散射检测器气体流量为1.6 L·min–1、漂移管温度为60 ℃、喷雾器温度为30 ℃。基于选定的过程样品，确定经发酵后含量升高的共有峰，进而建立淡豆豉特征图谱。以淡豆豉特征图谱为对照图谱，对15批市售淡豆豉样品的HPLC-ELSD色谱图进行相似度评价，并对其10个共有峰峰面积进行正交偏最小二乘法-判别分析评价。采用HPLC-二极管阵列检测器法（HPLC-DAD）对淡豆豉中6个大豆异黄酮类物质进行含量测定；采用HPLC-ELSD对淡豆豉中3个B族大豆皂苷进行含量测定。结果 基于10批30个发酵过程样品，发现淡豆豉中存在多个含量升高的色谱峰；其中10个共有峰峰面积升高较显著，进而建立了淡豆豉的特征图谱；通过与对照品比对，指认了其中6个共有峰。根据市售淡豆豉样品的HPLC-ELSD特征图谱相似度是否大于0.95，可将15批市售淡豆豉分为优质淡豆豉和基本合格淡豆豉。10个共有峰峰面积的正交偏最小二乘法-判别分析结果与相似度分析结果一致。15批市售品中大豆苷元和染料木素总质量分数为0.12%±0.06%，大豆皂苷Ba、大豆皂苷Bb和大豆皂苷Bc的总质量分数为0.59%±0.09%；优质淡豆豉中大豆苷元和染料木素总质量分数为0.15%±0.07%，大豆皂苷Ba、大豆皂苷Bb、大豆皂苷Bc总质量分数为0.63%±0.10%，大豆皂苷Bb质量分数为0.46%±0.08%；基本合格淡豆豉中大豆苷元和染料木素总质量分数为0.09%±0.04%，大豆皂苷Ba、大豆皂苷Bb、大豆皂苷Bc总质量分数为0.56%±0.07%，大豆皂苷Bb质量分数为0.41%±0.06%。结论 建立的HPLC-ELSD特征图谱精密度、重复性、稳定性良好，能够体现淡豆豉中大豆皂苷与异黄酮多种组分的含量特点，可用于提升淡豆豉的质量控制水平。指标性成分含量测定方面，建议在《中华人民共和国药典》2020年版标准基础上，进一步提高大豆苷元和染料木素总量的限量标准，且可增加大豆皂苷Bb含量测定项。 |
| 中文关键词:淡豆豉 质量标准提升 特征图谱 大豆异黄酮苷元 B族大豆皂苷 |
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| Establishment of HPLC-ELSD Specific Chromatogram of Sojae Semen Praeparatum Based on Fermentation Process and Content Determination of Index Components |
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| Abstract:Objective To investigate the changes in chemical components during the fermentation process of Sojae Semen Praeparatum (SSP), establish a specific chromatogram for quality evaluation using HPLC-ELSD, and evaluate the content of soy isoflavones and soyasaponins in SSP.Methods An Agilent ZORBAX SB C18 column (150 mm×4.6 mm, 3.5 μm) was used. The mobile phase A was 0.1% acetic acid-water, and the mobile phase B was 0.1% acetic acid-acetonitrile, with gradient elution at a flow rate of 1.0 mL·min–1. The column temperature was 30 ℃, and an evaporative light scattering detector (gas flow rate of 1.6 L·min–1, drift tube temperature of 60 ℃, sprayer temperature of 30 ℃) was employed. Based on the selected process samples, common peaks with elevated content were identified after fermentation, and a specific chromatogram for SSP was established. The HPLC-ELSD chromatograms of 15 commercial SSP samples were evaluated for similarity using the SSP specific chromatogram as a reference, and the peak areas of the 10 common peaks were assessed using orthogonal partial least squares-discriminant analysis. The content of 6 soybean isoflavones was determined by HPLC-DAD, and the content of soyasaponin Ba, Bb, and Bc was determined by HPLC-ELSD.Results Based on 30 samples from 10 batches during the fermentation process, multiple peaks with elevated levels were identified in SSP by HPLC-ELSD. A specific chromatogram for SSP was established based on 10 common peaks with significantly increased areas, and 6 common chromatographic peaks were designated after comparison with reference standards. The 15 commercial SSP samples were categorized into high-quality SSP and basically qualified SSP based on whether their similarity of HPLC-ELSD specific chromatogram was greater than 0.95. The results were consistent with the orthogonal partial least squares-discriminant analysis of the peak areas of the 10 common peaks. The total content of daidzein and genistein in the 15 batches of commercial samples was 0.12%±0.06%, and the total content of soyasaponin Ba, Bb, and Bc was 0.59%±0.09%. In high-quality SSP, the total content of daidzein and genistein was 0.15%±0.07%, the total content of soyasaponin Ba, Bb, and Bc was 0.63%±0.10%, and the content of soyasaponin Bb was 0.46%±0.08%. In basically qualified SSP, the total content of daidzein and genistein was 0.09%±0.04%, the total content of soyasaponin Ba, Bb, and Bc was 0.56%±0.07%, and the content of soyasaponin Bb was 0.41%±0.06%.Conclusion The established HPLC-ELSD specific chromatogram showed good precision, reproducibility, and stability, reflecting the content characteristics of various components in SSP, such as soyasaponins and soy isoflavones. This method can improve the quality control standards of SSP. It is recommended to further enhance the total limit standards of daidzein and genistein based on the current standards in Chinese Pharmacopoeia 2020 edition and to add the content determination item of soyasaponin Bb. |
| keywords:Sojae Semen Praeparatum quality standard enhancement specific chromatogram soy isoflavone aglycones group B soyasaponins |
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