HPLC-一测多评法同时测定大青叶中6种抗肿瘤活性成分的含量
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篇名: HPLC-一测多评法同时测定大青叶中6种抗肿瘤活性成分的含量
TITLE:
摘要: 目的:建立同时测定大青叶中6种抗肿瘤活性成分含量的方法。方法:采用高效液相色谱法。色谱柱为Waters XTERRA® MS C18,流动相为甲醇-0.1%甲酸水溶液(梯度洗脱),流速为1.0 mL/min,柱温为35 ℃,检测波长为254 nm,进样量为10 μL。以木犀草苷为内标,分别计算其与尿苷、腺苷、金丝桃苷、7-甲氧基香豆素、芹菜素的相对校正因子(RCF),通过RCF计算大青叶中上述5种抗肿瘤活性成分的含量,同时采用外标法测定这6种成分的含量,比较一测多评法和外标法的含量测定结果。结果:尿苷、腺苷、木犀草苷、金丝桃苷、7-甲氧基香豆素、芹菜素进样量检测线性范围分别为0.122 6~4.902、0.063 4~2.534、0.032 7~1.309、0.038 2~1.527、0.032 3~1.291、0.052 5~2.098 μg(r≥0.999 1);检测限分别为3.06、1.58、0.82、0.96、0.81、1.31 μg/mL,定量限分别为10.21、5.28、2.73、3.18、2.69、4.37 μg/mL;精密度、稳定性(24 h)、重复性试验的RSD<2%(n=6);加样回收率为96.45%~99.14%( RSD为0.8%~1.6%,n=6)。尿苷、腺苷、金丝桃苷、7-甲氧基香豆素、芹菜素的RCF分别为0.197、0.413、0.732、0.825、0.587。在不同试验条件下,RCF重现性良好。采用一测多评法和外标法的含量测定结果比较,差异无统计学意义(P>0.05)。结论:该方法操作简单快捷、结果准确可靠,可用于大青叶中尿苷、腺苷、木犀草苷、金丝桃苷、7-甲氧基香豆素和芹菜素6种抗肿瘤活性成分含量的同时测定。
ABSTRACT: OBJECTIVE: To establish a method for simultaneous determination of 6 antitumor active constituents in Folium isatidis. METHODS: HPLC method was adopted. The determination was performed on Waters XTERRA® MS C18 column with mobile phase consisted of methanol-0.1% formic acid (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 35 ℃, and detection wavelength was 254 nm. The injection volume was 10 μL. Using luteolin as internal standard, relative correction factors (RCF) of uridine, adenosine, hyperin, 7-methoxycoumarin and apigenin were calculated respectively. The contents of above 5 antitumor active components in F. isatidis were calculated by RCF. The contents of 6 components were determined by external standard method (ESM). The results of content determination by QAMS were compared with ESM. RESULTS: The linear range was 0.122 6-4.90 2 μg for uridine, 0.063 4-2.534 μg for adenosine, 0.032 7-1.309 μg for luteolin, 0.038 2-1.527 μg for hyperin, 0.032 3-1.291 μg for 7-methoxycoumarin, 0.052 5-2.098 μg for apigenin(r≥0.999 1). The limits of detection were 3.06, 1.58, 0.82, 0.96, 0.81, 1.31 μg/mL, respectively. The limits of quantification were 10.21, 5.28, 2.73, 3.18, 2.69, 4.37 μg/mL. RSDs of precision, stability (24 h) and reproducibility tests were less than 2% (n=6). The average recoveries were 96.45%-99.14% (RSDs were 0.8%-1.6%, n=6). RCFs of uridine, adenosine, hyperin, 7-methoxycoumarin and apigenin were 0.197, 0.413, 0.732, 0.825 and 0.587, respectively. RCFs repeatability was perfect under different experiment conditions. There was no significant difference between the quantitative results of the QAMS and ESM (P>0.05). CONCLUSIONS: QAMS method is simple, fast, accurate and feasible, and can be used for simultaneous determination of uridine, adenosine, luteolin, hyperin, 7-methoxycoumarin and apigenin in F. isatidis.
期刊: 2018年第29卷第19期
作者: 罗文艳,段和祥,刘绪平,陈希,章瑛,李志勇
AUTHORS: LUO Wenyan,DUAN Hexiang,LIU Xuping,CHEN Xi,ZHANG Ying,LI Zhiyong
关键字: 高效液相色谱法;一测多评法;大青叶;木犀草苷;抗肿瘤活性成分;相对校正因子
KEYWORDS: HPLC; QAMS; Folium isatidis; Luteolin; Antitumor active constituent; Relative correction factor
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