Vitamin B1 (Thiamin)
Easy to use microbiological microtiter plate test for the quantification of total vitamin B1 (enriched and natural vitamin B1) in food, feed and pharmaceutical products.
Vitamin B1 is extracted from the sample and the extract is diluted. The diluted extract and the vitamin B1-assay-medium are pipetted into the wells of a microtiter plate which is coated with Lactobacillus fermentum. The growth of Lactobacillus fermentum is dependent on the supply of vitamin B1. Following the addition of vitamin B1 as a standard or as a compound of the sample, the bacteria grows 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 vitamin B1 is measured as turbidity and compared to a standard curve. The measurement is done using a 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)|
|Standard Range||0.012 – 0.060 mg / 100 g (ml)|
|Sample Preparation||For the determination of free vitamin B1 in nutrient-enriched samples, a hot acid extraction is usually sufficient. However, enriched drinks can be used directly after dilution. Due to the hot water extraction, natural vitamin B1 is only measured in part. For the determination of the total vitamin B1content, the sample has to be hydrolyzed with enzyme.|
|Incubation Time||44-48 h in the dark at 37 °C (98.6 °F)|