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目的:采用HPLC法建立风芍六君子汤水煎液的指纹图谱,并对其主要的指标性成分没食子酸、芍药苷和橙皮苷进行含量测定,为其成分结构分析和质量控制提供研究基础。方法:采用Diamonsil钻石C_(18)(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.2%磷酸水溶液为流动相,梯度洗脱(0~6 min,5% A;6~12 min,5% A→15% A;12~35 min,15% A→35% A;35~40 min,35% A→55% A;40~65 min,55% A→75% A;65~80 min,75% A→100% A;80~85 min,100% A),流速1.0 mL·min~(-1),柱温35℃,检测波长230 nm,采用《中药色谱指纹图谱相似度评价系统软件(2004A版)》,确定共有峰,计算相似度,并对其色谱峰进行了指认和归属。结果:建立了风芍六君子汤水煎液的指纹图谱,并对其方法学进行了验证,确定了指纹图谱中的19个共有峰,其相似度大于0.95,共有峰的相对保留时间及相对峰面积的RSD均小于3%,归属了其中的18个峰,并指认了没食子酸、对羟基苯甲醛等10个色谱峰的化学结构;没食子酸、芍药苷、橙皮苷的含量分别为3.819、9.053和6.558 mg·g~(-1)。结论:风芍六君子汤水煎液HPLC指纹图谱的精密度、稳定性和重复性良好,峰的指认归属准确,能够有效的控制其质量。
OBJECTIVE: To establish the fingerprint of Fengshuiliujunzitang decoction by HPLC and to determine the content of its main constituents, gallic acid, paeoniflorin and hesperidin, and to provide the basis for its structural analysis and quality control. METHODS: Diamonsil diamond column was used to elute gradient C018 (250 mm × 4.6 mm, 5 μm) column with acetonitrile-0.2% phosphoric acid as mobile phase (0-6 min, 5% A, 6-12 min, 5% A → 15% A; 12-35 min, 15% A → 35% A; 35-40 min, 35% A → 55% A; 40-65 min, 55% A → 75% A; 65-80 min, 75% A → 100% A; 80-85 min, 100% A) at a flow rate of 1.0 mL · min -1 .The column temperature was 35 ℃ and the detection wavelength was 230 nm. System Software (Version 2004A) "to identify common peaks, calculate the similarity, and identify and assign their peaks. Results: The fingerprint of Fengsha Liujunzi decoction was established and its methodology was validated. The 19 common peaks in the fingerprint were identified, the similarity was greater than 0.95, the relative retention time and the relative peak of the common peak The RSDs of the areas were all less than 3%, and 18 peaks were assigned to them. The chemical structures of 10 peaks such as gallic acid and p-hydroxybenzaldehyde were also identified. The contents of gallic acid, paeoniflorin and hesperidin were 3.819, 9.053 and 6.558 mg · g -1. Conclusion: The precision, stability and repeatability of HPLC fingerprints of Fengsha Liujunzi Decoction decoction are good, the identification of peak is accurate, and its quality can be effectively controlled.