龙胆药材中龙胆苦苷、马钱酸含量测定与其外观性状的相关性及质量等级标准研究
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篇名: 龙胆药材中龙胆苦苷、马钱酸含量测定与其外观性状的相关性及质量等级标准研究
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摘要: 目的:建立同时测定龙胆药材中龙胆苦苷、马钱酸含量的方法,并考察其含量与外观性状的相关性及进行质量等级划分。方法:采用高效液相色谱法。色谱柱为Ascentis Express C18,流动相为0.1%磷酸水溶液-乙腈(梯度洗脱),流速为0.4 mL/min,柱温为30 ℃,检测波长为240 nm,进样量为1 μL。以药材长度、须根数及根直径为指标,考察龙胆药材的外观性状特征;采用SPSS 21.0软件分析药材中龙胆苦苷、马钱酸含量与其外观性状的相关性;采用SPSS 21.0软件进行k-均值聚类分析,并建立龙胆药材的质量等级划分标准。结果:龙胆苦苷、马钱酸检测质量浓度线性范围分别为0.5~3.0 mg/mL(r=0.999 9)、0.05~0.50 mg/mL(r=0.999 9);定量限分别为0.295、0.289 μg/mL,检测限分别为0.082、0.081 μg/mL;精密度、稳定性、重复性试验的 RSD均小于2%;加样回收率分别为97.56%~102.23%(RSD=1.56%,n=6)、97.58%~102.67%(RSD=1.86%,n=6)。相关性分析结果显示,药材中龙胆苦苷、马钱酸含量与其长度、须根数、根直径均呈正相关,且对含量的影响程度大小依次为须根数>长度>根直径。聚类分析结果显示,54批龙胆药材可分聚为2类,S4~S6、S13、S17~S23、S25、S28、S31~S34聚为一类;S1~S3、S7~S12、S14~S16、S24、S26、S27、S29、S30、S35~S54聚为一类。质量等级划分结果显示,54批龙胆药材可分为2个等级,其结果与聚类分析结果一致。结论:所建含量测定方法操作简便,稳定性较好,可用于同时测定龙胆药材中龙胆苦苷、马钱酸的含量;龙胆药材须根数越多、长度越长、根直径越粗,马钱酸和龙胆苦苷的含量越高,药材质量越好。
ABSTRACT: OBJECTIVE: To establish the method for the content determination of gentiopicrin and loganic acid in Gentiana scabra, and to investigate the correlation of their contents with appearance traits and quality gradation criterion. METHODS: HPLC method was adopted. The determination was performed on Ascentis Express C18 column with mobile phase consisted of 0.1% phosphoric acid-acetonitrile (gradient elution) at the flow rate of 0.4 mL/min. The column temperature was 30 ℃, and detection wavelength was set at 240 nm. The sample size was 1 μL. Taking the length, number and diameter of fibrous roots as indexes, the appearance and morphological characteristics of G. scabra were studied. The relationship of gentiopicrin and loganic acid content with the appearance property of medicinal material was analyzed by SPSS 21.0 software. k-mean clustering analysis was carried out by using SPSS 21.0 software, and gradation standard for G. scabra was established preliminarily. RESULTS: The linear range of gentiopicrin and loganic acid were 0.5-3.0 μg/mL (r=0.999 9) and 0.05-0.50 μg/mL (r=0.999 9). The limit of quantification of gentiopicrin and loganic acid were 0.295, 0.289 μg/mL; the detection limit were 0.082, 0.081 μg/mL; RSDs of precision, stability, repeatability tests were all lower than 2%; the recovery rates were 97.56%- 102.23% (RSD=1.56%, n=6) and 97.58%-102.67% (RSD=1.86%, n=6). Correlation results showed that there was a significant positive correlation of the length of G. scabra, the number of roots, root diameter, with the contents of gentiopicrin and loganic acid. The order of affecting content was the number of roots >length >root diameter. k-means clustering analysis showed that 54 batches of G. scabra was divided into two categories; S4-S6,S13,S17-S23,S25,S28,S31-S34 were clustered into a category; S1-S3, S7-S12, S14-S16, S24, S26,S27,S29, S30,S35-S54 were clustered into the other category. The results of gradation showed that 54 batches of G. scabra could be divided into two grades, and the results were consistent with the cluster analysis. CONCLUSIONS: Established method is simple and stable, and can be used for simultaneous determination of gentiopicrin and loganic acid in G. scabra. The more fibrous roots, the longer the length, the thicker the root, the higher the content of gentiopicroside and loganic acid, the better the quality of G. scabra.
期刊: 2019年第30卷第16期
作者: 刘战,侯晓琳,宿莹,吴晓燕,孙金,姜雨昕,翁丽丽
AUTHORS: LIU Zhan,HOU Xiaolin,SU Ying,WU Xiaoyan,SUN Jin,JIANG Yuxin,WENG Lili
关键字: 龙胆;龙胆苦苷;马钱酸;高效液相色谱法;外观性状;相关性;聚类分析;质量等级标准
KEYWORDS: Gentiana scabra; Gentiopicrin; Loganic acid; HPLC; Appearance trait; Correlation; Cluster analysis; Quality gradation criterion
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