UPLC特征图谱结合黄酮类活性成分含量测定的沙棘配方颗粒质量研究

作者：蔡小兵1，2，3，江 斌1，2，3，谭 沛1，2，3

单位：1.华润三九现代中药制药有限公司，广东 惠州 516000； 2.中药配方颗粒安徽省重点实验室，安徽 淮北 235000； 3.华润三九医药股份有限公司，广东 深圳 518000

引用：引用：蔡小兵，江斌，谭沛.UPLC特征图谱结合黄酮类活性成分含量测定的沙棘配方颗粒质量研究[J].中医药导报,2025,31(1):61-66.

DOI：10.13862/j.cn43-1446/r.2025.01.011

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摘要：目的：建立沙棘配方颗粒质量评价方法，整体评价不同产地沙棘配方颗粒产品质量的均一性及差异化。方法：采用UPLC法，色谱柱为Shim-pack GIST C18-AQ HP（150.0 mm×2.1 mm，1.9 μm），流动相为乙腈（A）-0.05%磷酸水溶液（B），梯度洗脱，流速为0.3 mL/min，检测波长为260 nm，柱温为30 ℃，对不同产地沙棘配方颗粒进行相似度评价、聚类分析（CA）、主成分分析（PCA）及正交偏最小二乘判别分析（OPLS-DA），并采用相同UPLC色谱系统及方法同时测定多种黄酮类成分含量，对18批沙棘配方颗粒进行质量分析和评价。结果:沙棘配方颗粒特征图谱确定12个共有峰且全部指认，分别为峰1（5-羟基糠醛）、峰2（原儿茶酸）、峰3（槲皮素-3-O-槐二糖-7-O-鼠李糖苷）、峰4（异鼠李素-3-O-槐二糖-7-O-鼠李糖苷）、峰5（异鼠李素-3-O-葡萄糖-7-O-鼠李糖苷）、峰6（芦丁）、峰7（异槲皮苷）、峰8（水仙苷）、峰9（异鼠李素-3-O-葡萄糖苷）、峰10（田基黄苷）、峰11（槲皮素）、峰12（异鼠李素）。18批沙棘配方颗粒特征图谱与对照图谱相似度均大于0.90，聚类分析将全部样品分为2类，两个主成分的累积方差贡献率为92.433%，并筛选出3个影响质量的差异性成分；芦丁、异槲皮苷、水仙苷、田基黄苷、槲皮素、异鼠李素在各自质量浓度范围内线性关系良好；仪器精密度、方法重复性、溶液稳定性试验RSD值均＜3%。结论：不同产地沙棘样品存在一定的质量差异。本研究建立的特征图谱及黄酮类指标成分含量测定方法可客观、全面、准确地对沙棘配方颗粒进行质量评价。

关键词：沙棘配方颗粒；UPLC法；聚类分析；质量研究

Abstract：Objective: To establish a quality evaluation method for seabuckthorn formula granules, and to evaluate the uniformity and differentiation of seabuckthorn formula granules from different producing areas. Methods: UPLC chromatography was performed on Shim-pack GISTC18-AQ HP (150.0 mm×2.1 mm, 1.9 μm) column with mobile phase consisting of acetonitrile (A) -0.05% phosphoric acid aqueous solution (B) with gradient elution at the flow rate of 0.3 mL/min. The detection wavelength was 260 nm and column temperature was 30 ℃. The similarity evaluation, cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were carried out for seabuckthorn formula granules from different origin. The same UPLC chromatography system and method were used to determine the content of multiple flavonoids, and the quality of 18 batches of seabuckthorn formula granules was analyzed and evaluated. Results: A total of 12 common peaks were identified and all of them were identified by the characteristic map of seabuckthorn formula granules. They were peak 1 (5-hydroxyfurfural), peak 2 (protocatechuic acid), peak 3 (quercetin 3-O-sophorobiose-7-O-rhamnoside), peak 4 (isorhamnose-3-O-glucose-7-o-rhamnoside), peak 5 (isorhamnose-3-o-glucose-7-o-rhamnoside), peak 6 (rutin), peak 7 (isoquercetin), peak 8 (narcissoside), peak 9 (isorhamnein-3-O-glucoside), peak 10 (chrysanthin), peak 11 (quercetin) and peak 12 (isorhamnetin). The similarity between the characteristic maps of 18 batches of seabuckthorn formula granules and the control maps was greater than 0.90. All samples were divided into two categories by cluster analysis. The cumulative variance contribution rate of the two principal components was 92.433%, and three different components affecting the quality were screened out. The linear relationship of rutin, isoquercetin, narcissoside, chrysanthin, quercetin and isorhamnetin showed good linear relationships within the concentration range. RSD of instrument precision, method repeatability and solution stability tests were all lower than 3%. Conclusion: There are certain quality differences in seabuckthorn samples from different producing areas. The specific chromatogram and the method of flavonoid index component content determination established in this study can objectively, comprehensively and accurately evaluate the quality of seabuckthorn formula granules.

Key words：seabuckthorn formula granule; UPLC chromatography; cluster analysis; quality study

发布时间：2025-11-28

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