保元颗粒分时段多波长融合指纹图谱及 7个指标性成分含量测定

Determination of the multi-wavelength fusion fingerprint and seven marker components in Bao Yuan granules at different time periods

摘要

目的 建立保元颗粒高效液相色谱法的分时段多波长融合指纹图谱，构建其主要功效成分的含量测定方法。方法 采用高效液相色谱法-二极管阵列检测法全波长扫描构建10批保元颗粒的指纹图谱，基于该图谱中所包含的主要色谱峰，测定保元颗粒中人参皂苷Rg ₁、人参皂苷Re、人参皂苷Rb ₁、毛蕊异黄酮苷、甘草苷、6-姜辣素、桂皮醛7个指标性成分的含量。结果 指纹图谱测定的色谱条件为色谱柱Spursil C ₁₈（250 mm×4.6 mm, 5 µm）；流动相乙腈-水，梯度洗脱；流速0.8 mL/min；柱温25°C；选择203、237、260、280、290 nm共5个波长获得融合指纹图谱。10批样品的融合指纹图谱包含29个共有峰，分别来自保元颗粒组方中的不同药材，其中7个共有峰来自人参，15个共有峰来自黄芪，11个共有峰来自甘草，1个共有峰来自肉桂，8个共有峰来自生姜。10批保元颗粒相似度为0.878~0.985。7个指标成分的标准曲线相关系数 R ²均大于0.990 0，线性良好。方法学考察发现，构建的指纹图谱和7个指标成分含量测定方法的精密度、重复性、稳定性相对标准偏差均小于5%，7个指标成分加样回收率为97.21%~103.10%。结论 构建的多波长融合指纹图谱能较全面地反映保元颗粒的整体信息，还可用于组方药味的鉴别，为保元颗粒质量标准研究提供了方法学基础。

Abstract

Objective To establish a multi-wavelength fusion fingerprint of Baoyuan Granules by high performance liquid chromatography at different time periods, and develop a method for determining the content of its main active ingredients. Methods The fingerprints of 10 batches of Baoyuan Granules were constructed through high performance liquid chromatography-diode array detector full-wavelength scanning. Based on the identification of the main chromatographic peaks in these fingerprints, the contents of seven indicator components in Baoyuan Granules, namely ginsenoside Rg ₁, ginsenoside Re, ginsenoside Rb ₁, calycosin-7-O-β-D-glucoside, liquiritin, 6-gingerol and cinnamaldehyde were determined. Results The chromatographic conditions for fingerprint determination were that the chromatographic column was Spursil C ₁₈ (250 mm×4.6 mm, 5 µm), the mobile phase was acetonitrile-water with gradient elution, the flow rate was 0.8 mL/min and the column temperature was 25 °C. A fusion fingerprint was obtained by selecting five wavelengths (203, 237, 260, 280, and 290 nm). The fusion fingerprints of the 10 batches of samples contained 29 common peaks, which originated from different medicinal materials in the formula of Baoyuan Granules. Specifically, 7 common peaks were from ginseng, 15 from astragalus membranaceus, 11 from licorice, 1 from cinnamon and 8 from ginger. The similarity of the 10 batches of Baoyuan Granules was between 0.878 and 0.985. The correlation coefficients R ² of the standard curves of the seven indicator components were all above 0.990 0, indicating a good linearity. The methodological investigation showed that the relative standard deviations of precision, repeatability, and stability of the constructed fingerprint and the content determination methods of the seven indicator components were all less than 5%. The spiked recoveries of the seven indicator components were between 97.21% and 103.10%. Conclusion The multi-wavelength fusion fingerprint constructed could not only comprehensively reflect the overall information of Baoyuan Granules, but also be used to identify the medicinal materials in the formula. This method provides a methodological basis for the research on the quality standard of Baoyuan Granules.

关键词

保元颗粒 / 高效液相色谱 / 指纹图谱 / 含量测定 / 多波长融合 / 组方药

Key words

Bao Yuan granules / high performance liquid chromatography / fingerprint / content determination / Multi-wavelength fusion / formula

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基金资助

甘肃省中医药科研课题(GZKZ-2024-30)

兰州市科技计划项目(2024-2-24)

兰州市科技计划项目(2024-8-1)