基于对照制剂的板蓝根颗粒指纹图谱和多成分含量测定研究及质量等级初评价

Research of chromatographic fingerprint and the determination of multiple constituents and the primary evaluation of the quality grade of Banlangen granules based on the reference drug

目的:以对照制剂为随行对照,分别建立指纹图谱和多成分指标含量测定方法,对55个厂家103批板蓝根颗粒质量进行分级评价。方法:采用超高效液相色谱法,建立板蓝根颗粒指纹图谱,同时测定7个成分(尿苷、腺嘌呤、鸟苷、(R,S)-告依春、腺苷、紫丁香酚苷、直铁线连宁B)含量。采用Waters ACQUITY UPLC HSS T_(3)(100 mm×2.1 mm,1.8μm)色谱柱,以甲醇-水为流动相,梯度洗脱,流速0.2 mL·min^(-1),进样量2μL,柱温30℃,采用多波长检测模式。与对照品比较,确定特征峰及其化合物归属。以对照制剂为参照,计算样品指纹图谱相似度及成分含量,并初步分级评价其质量。结果:建立了板蓝根颗粒特征指纹图谱,共指认10个特征峰,样品指纹图谱相似度在0.541~0.993范围,101批制剂相似度>0.75,达到二级限度,81批制剂相似度≥0.90,达到一级限度。7个成分在各自范围内线性关系良好,平均回收率(n=9)为97.0%~104.7%(RSD均<3%),精密度、稳定性、重复性良好(RSD均<3%)。以5 g·袋^(-1)计,7个成分总含量范围为0.189~10.347 mg·袋^(-1)。结果显示,75批次样品的含量达到二级限度,59批次样品的含量达到一级限度。结论:建立的方法简便、准确、快速,可用于板蓝根颗粒的质量控制和等级评价,也可为其他中成药质量等级评价研究提供参考。

Objective:To establish the determination methods for chromatographic fingerprint and multiple constituents,and then evaluate the quality grade of 103 batches of Banlangen granules that were from 55 manufacturers based on the reference drug.Methods:Ultra-high-performance liquid chromatography(UPLC)analysis was used to establish the fingerprint of Banlangen granules and quantitate the contents of 7 components-uridine,adenine,vernine,(R,S)-goitrin,adenosine,syringin and clemastanin B.The test was performed on the Waters ACQUITY UPLC HSS T_(3)(100 mm×2.1 mm,1.8μm)column with gradient elution using methanol-water at a flow rate of 0.2 mL·min^(-1).The injection volume was 2μL,and the column temperature was constant at 30℃.Then,the results were detected using adopting a multi-wavelength detection mode.Next,the characteristic peaks and their belonging were clarified by comparing them with the reference drug.The sample fingerprint similarity and determination of contents were finally calculated,and the quality grades of Banlangen granules in each item were evaluated by using the reference drug.Results:The fingerprint was established,and 10 main characteristic peaks were identified.The similarities of the sample's fingerprints ranged from 0.541 to 0.993,of which 101 batches were over 0.75 that were reached the second level,and 81 batches were over 0.90 that were reached the first level.The linear ranges of 7 components had good linear relationships within their respective ranges.The average recoveries(n=9)were 97.0%-104.7%with RSDs all below 3%.The precision,stability,and repeatability of methods were all good with RSDs all below 3%.Unified the sample specification to 5 g per bag,the content range of Banlangen granules samples was between 0.189-10.347 mg per bag.As a result,the content of 75 batches of samples reached the second level,and the content of 59 batches of samples reached the first level.Conclusion:The established methods are simple,accurate and rapid,which can be used for the quality control and grade evaluation of Banlangen granules,as well as the research paradigm of other traditional Chinese medicine preparations.