α-Ribazole, a fluorescent marker for the liquid chromatographic determination of vitamin B12 in foodstuffs

被引:38
作者
Pakin, C
Bergaentzlé, M
Aoudé-Werner, D
Hasselmann, C
机构
[1] Fac Pharm, Lab Chim Analyt & Sci Aliment, UMR 7512, F-67401 Illkirch Graffenstaden, France
[2] Aerial, F-67412 Illkirch Graffenstaden, France
关键词
food analysis; vitamin B-12; alpha-ribazole; liquid chromatography;
D O I
10.1016/j.chroma.2005.05.066
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A method to determine the contents of free vitamin B-12 in various foods by reversed phase liquid chromatography-fluorimetry is reported. It includes a purification of the samples by passage through an immunoaffinity column and a pre-column conversion of vitamin B-12 into the fluorescent alpha-ribazole (successive treatments of the extract with 2.5 M sodium hydroxide (at 100 degrees C for 15 min) and alkaline phosphatase (7.5 U) at 37 degrees C and pH 8 for 16 h). An enzymatic hydrolysis prior to the purification step (pepsin at 37 degrees C and pH 4 for 3 h) made it possible to release the vitamin B-12 bound to proteins and thus to obtain the total vitamin B-12 contents of these foodstuffs. The method proposed for the determination of free and bound vitamin B-12 gives a good recovery rate (95-100%) and a satisfactory repeatability (R.S.D.r between 1.0 and 5.4%). Owing to its low quantification limit (3 ng g(-1)) and the good resolution of the alpha-ribazole peak, it could most probably be applied to the determination of this vitamin in any foodstuff. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:182 / 189
页数:8
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