Summary - biotin

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EN 15607:2009 Foodstuffs - Determination of d-biotin by HPLC:


This European Standard specifies a method for the determination of the mass fraction of d-biotin by high performance liquid chromatography (HPLC). The method has been validated in an inter-laboratory test on fortified and non-fortified samples such as cereal breakfast powder, infant milk powder, lyophilized green peas with ham, lyophilized chicken soup and on nutritive orange juice, at levels from 16 µg/100 g to 200 µg/100 g.

NOTE 1 d-biocytin can also be estimated by this method. But none of the samples used for the validation step contained d-biocytin. Nonetheless the recovery rate is more than 90 % for d-biotin and d-biocytin.

NOTE 2 The method underestimates the real biotin content when used for samples containing egg.


D-biotin is extracted from food after enzymatic treatment and quantified by HPLC with post-column binding reaction.

The complexion of d-biotin with avidin appears to be very specific. On that account, this protein, covalently bound to a fluorescent marker, fluorescein 5-isothiocyanate, can be used as a reagent for a post-column derivatisation of d-biotin.

Precision data