文章基本信息

标题：酸性ホスファターゼによるリボフラビンへの変換に基づく前処理法を利用したリン酸リボフラビンナトリウムのHPLC定量
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作者：大曽根圭子 ; 上野清一 ; 石崎睦雄等
期刊名称：Journal of Health Science
印刷版ISSN：1344-9702
电子版ISSN：1347-5207
出版年度：1995
卷号：41
期号：5
页码：358-363
DOI：10.1248/jhs1956.41.358
出版社：The Pharmaceutical Society of Japan
摘要：A simple and rapid method for high-performance liquid chromatographic determination of riboflavin sodium phosphate (FMN) based on its conversion to riboflavin (FR) with acid phosphatase was developed. A mixture for the conversion of FMN to FR contained, in a final volume of 4.0 ml, 1.0 ml of 0.8 M sodium acetate buffer (pH 4.8), 20.0 units of acid phosphatase (Type I : from wheat germ, Sigma Chemical Co.) and less than 0.5 mg of FMN. After incubation for 30 min at 37°C in an amber test tube with a stopper, the reaction mixture was filtered through a 0.45 μm membrane filter. The filtrate was injected to reversed-phase high performance liquid chromatography equipped with an octadecylsilyl column (L-column ODS, 4.6 mm i.d.×250 mm) and detected for absorbance at 270 nm. FR in the acidic filtrate was stable for at least 24 h. The recoveries of FMN in drugs and soft drinks by this method were 97.7-100.8% with the coefficients of variation of 0.4-2.8%. Impurity FR was detected in all samples under the same conditions without enzymatic hydrolysis. This method could be applied to the separative determination of FR and FMN.
关键词：riboflavin sodium phosphate;riboflavin;acid phosphatase;enzymatic hydrolysis;HPLC;quantitative analysis