Easy to use microbiological microtiter plate test for the quantification of total biotin (enriched and natural biotin) in food, feed and pharmaceutical products.
Biotin is extracted from the sample and the extract is diluted. The diluted extract and the biotin assay-medium are pipetted into the wells of a microtiter plate which is coated with Lactobacillus plantarum. The growth of Lactobacillus plantarum is dependent on the supply of biotin. Following the addition of biotin as a standard or as a compound of the sample, the bacteria grow until the vitamin is consumed. The incubation is done in the dark at 37 °C (98.6 °F) for 44 – 48 h. The intensity of metabolism or growth in relation to the extracted biotin is measured as turbidity and compared to a standard curve. The measurement is done using an microtiter plate spectrophotometer at 610 – 630 nm (alternatively at 540 – 550 nm).
|Test format||Microtiter plate with 96 wells (12 strips with 8 removable wells each)|
|Incubation time||44 - 48 h in the dark at 37 °C (98.6 °F)|