根皮苷候选化学对照品的制备及分析研究

Preparation and analysis of candidate chemical reference substance of phlorizin

目的研究根皮苷候选化学对照品的制备方法及其质量标准,为含有根皮苷的药材及其相关产品的质量控制提供参考。方法多穗柯叶片粉碎,用70%的乙醇溶液(V/V)提取,提取液滤过,浓缩成稠膏,用水稀释,用石油醚(60~90℃)洗涤2次,再用乙酸乙酯提取2次,收集乙酸乙酯部分,采用硅胶柱色谱以二氯甲烷-甲醇系统作为流动相梯度洗脱,收集含有根皮苷的流份,合并,浓缩,再用反相制备高效液相色谱法,以甲醇-水(40:60,V:V)为流动相,以紫外光284 nm作为检测波长,进行分离纯化,所得的流份经甲醇重结晶得到高纯度根皮苷,通过紫外光谱、红外光谱、高分辨质谱、核磁共振氢谱、核磁共振碳谱表征鉴定其化学结构。采用高效液相色谱法和质量平衡法测定并计算其纯度。结果制备的根皮苷候选化学对照品经高效液相色谱纯度测定均值为98.59%,炽灼残渣测定均值为0.02%,以质量平衡法计算含量为98.48%。结论制备得到的高纯度根皮苷对照品符合中药化学对照品技术要求,可作为相关产品质量控制的候选对照品;建立的分析方法准确、可靠,可为对照品质量控制提供科学依据。

Objective To study the preparation method and quality standard of phloridzin,and to provide reference for the quality control of medicinal materials containing phloridzin and its related products.Method The leaves of Lithocarpus litseifolius were smashed extracted with 70%ethanol solution(V/V),filtered and concentrated into thick paste,diluted with water,washed twice with petroleum ether(60-90℃),and then extracted twice with ethyl acetate.The ethyl acetate fraction was collected and eluted by silica gel column chromatography with dichloromethane methanol system as mobile phase.Separation was performed by high performance liquid chromatography(HPLC),the mobile phase was methanol-whter(40:60,V:V)at 284 nm.High purity phloridzin was obtained by recrystallization with methanol.Identification of chemical structure by ultraviolet,infrared radiation,high resolution mass,Nuclear magnetic resonance hydrogen spectrum,nuclear magnetic resonance carbon spectrum.Results The average purity of phloridzin was 98.59%by HPLC,and the mean value of ash was 0.02%.The content of phlorizin was 98.48%by mass balance method.Conclusion The prepared high-purity phloridzin reference material meets the technical requirements of traditional Chinese medicine chemical reference material,and can be used as the candidate reference substance for quality control of related products.The established analytical method is accurate and reliable,which can provide scientific basis for the quality control of reference substance.