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目的:建立马海治瘫胶囊的指纹图谱,并测定其中主要成分的含量,为其制剂工艺的稳定性及质量控制提供科学依据。方法:采用Inertsil ODS-3色谱柱,以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱,检测波长为250 nm(0~23 min和31~120 min)、230 nm(23~31 min),柱温为30 ℃,流速为1.0 mL/min。应用“中药色谱指纹图谱相似度评价软件”(2012版)建立10批马海治瘫胶囊的高效液相色谱(HPLC)指纹图谱并进行相似度评价,分别以对照药材、缺味药材阴性样品和对照品对色谱峰进行归属和指认,并对指认出的主要成分进行定量分析。结果:10批样品的相似度均在0.99以上,共确定了20个共有峰,并指认了其中10个色谱峰。其中,1、13、14、15、16、17、18、19、20号色谱峰来源于大黄,3、4、6、7号色谱峰来源于制马钱子,8号色谱峰来源于当归,2、5、9、10、11、12号色谱峰未发现相应的药材来源;通过与对照品比对,指认了1、4、6、7、8、16、17、18、19、20号色谱峰分别为没食子酸、马钱苷酸、士的宁、马钱子碱、阿魏酸、芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚。在对指认出的5个主要成分(马钱苷酸、士的宁、马钱子碱、大黄素和大黄酚)的含量测定中,方法学考察均符合相关标准,10批样品中马钱苷酸、士的宁、马钱子碱、大黄素和大黄酚含量分别为2.477 1~2.785 9、1.746 1~1.946 0、1.374 6~1.505 8、1.573 2~1.824 1和0.232 1~0.261 7 mg/g。结论:建立的方法可靠、准确、稳定、简便,可作为马海治瘫胶囊的制剂工艺与质量监控依据。
OBJECTIVE: To establish the fingerprint of Mahai zhitan capsule, to determine the contents of main components, and to provide scientific basis for the stability and quality control of the preparation technology. METHODS: The determination was performed on Inertsil ODS-3 column with acetonitrile-0.1% phosphoric acid as mobile phase (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 250 nm (0-23 min and 31-120 min) and 230 nm (23-31 min). The column temperature was set at 30 ℃. HPLC fingerprint for 10 batches of Mahai zhitan capsule was established by using “similarity evaluation software for chromatographic fingerprint of traditional chinese medicine” (2012 edition) and the similarity was evaluated. The chromatographic peaks were assigned and identified with reference substance, negative samples without ingredient and substance control respectively, and the identified main components were quantitatively analyzed. RESULTS: The similarity of 10 batches of sample was more than 0.99; 20 common peaks were found, and 10 common peaks were identified. Among them, No. 1,13,14,15,16,17,18,19,20 chromatographic peaks originated from Rheum palmatum; No. 3,4,6,7 chromatographic peaks originated from processed Strychnos nuxvomica; No. 8 chromatographic peaks originated from Angelica sinensis; the corresponding source of medicinal materials was not found in No. 2,5,9,10,11,12 chromatographic peaks. By comparing the reference substances, No. 1,4,6,7,8,16,17,18,19 and 20 chromatographic peaks were identified as gallic acid, loganin acid, strychnine, brucine, ferulic acid, aloe-emodin, rhein, emodin, chrysophanol and emodin methyl ether, respectively. In the determination of identified five main components (loganin, strychnine, brucine, emodin and chrysophanol), the methodological investigation met the relevant standards. In 10 batches of samples, the contents of loganin, strychnine, brucine, emodin and chrysophanol were 2.477 1-2.785 9, 1.746 1-1.946 0, 1.374 6-1.505 8, 1.573 2-1.824 1 and 0.232 1-0.261 7 mg/g, respectively. CONCLUSIONS: The established method is reliable, accurate, stable and simple, which could provide reference for the preparation technology and quality control of Mahai zhitan capsule.
马海治瘫胶囊指纹图谱含量测定高效液相色谱法
Mahai zhitan capsuleFingerprintContent determinationHPLC
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