注射用灰树花倍他葡聚糖的高效液相色谱指纹图谱研究

目的采用高效液相色谱法建立注射用灰树花倍他葡聚糖的指纹图谱。方法采用Shodex OH pak SB-804HQ色谱柱,以0.2 mol/L磷酸二氢钠溶液为流动相,流速0.5 mL/min,柱温35℃,以示差折光检测器(RID)进行检测,建立反映药材、中间体和注射剂多糖活性成分的指纹图谱(1);采用Agilent Eclipse XDB-C18色谱柱,以乙腈为流动相A,水(含0.05%磷酸)为流动相B,进行梯度洗脱,流速1.0 mL/min,柱温30℃,以二极管阵列检测器(DAD)在波长254 nm处进行检测,建立反映药材其它成分指纹特性的指纹图谱(2)。结果10批灰树花发酵菌丝体药材、中间体和注射剂的相似度均在95%以上,多糖活性成分在药材、中间体和注射剂之间表现出良好的相关性,方法学考察结果也符合要求。结论建立的指纹图谱既体现了多糖活性成分的相关性,又体现了药材其它组分的指纹特征,可用于灰树花药材鉴定和注射剂的质量控制。

Fingerprint of Grifola frondosa beta-glucan for injection by HPLC

Purpose To establish HPLC method for fingerprint determination of Grifola frondosa beta-glucan for injection(GFGI).Methods Chromatographic conditions of fingerprint including Grifola frondosa medicinal material,intermediate and GFGI(1) were as follows:Shodex OH pak SB-804HQ column,0.2 mol/L NaH2PO4 as mobile phase,flow rate was 0.5 mL/min,temperature at 35℃ and the detector was refractive index detector(RID).Chromatographic conditions of medicinal materials fingerprint(2) were available as follows: Agilent Eclipse XDB-C18 column,acetonitrile and water including 0.05% phosphate acid as mobile phase,flow rate was 1.0 mL/min,gradient elution,temperature at 30℃,and the detector was Diode Array Detector(DAD) at 254 nm.Results Similarity of 10 batches of liquid-cultured Grifola frondosa medicinal materials,intermediates and preparations were over 95%.There is good relativity among the medicinal material,intermediate and preparation.This method had a good precision,repeatability and reproducibility.Conclusion The analytical methods of fingerprint reflected not only the relativity of active ingredient among the medicinal material,intermediate and preparation,but also the fingerprint characteristics of Grifola frondosa medicinal material.It can be used for identification of Grifola frondosa medicinal material and quality control of GFGI.