Vitamin B5 (Pantothenic Acid)
Easy to use microbiological microtiter plate test for the quantification of total pantothenic acid (enriched and natural pantothenic acid) in food, feed and pharmaceutical products.
Pantothenic acid is extracted from the sample and the extract is diluted. The diluted extract and the pantothenic acid 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 pantothenic acid. Following the addition of pantothenic acid 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 20 – 24 h. The intensity of metabolism or growth in relation to the extracted pantothenic acid 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)|
|Standard Range||0.04 – 0.24 mg / 100 g (ml)|
|Sample Preparation||For the determination of added pantothenic acid in nutrient-enriched samples, a hot acid extraction is usually sufficient. However, enriched liquid samples can be used directly after sterile filtration and dilution. Due to the hot water extraction, natural pantothenic acid is only measured in part. For the determination of the total pantothenic acid content Native and added), the sample has to be treated with enzyme.|
|Incubation Time||20 – 24 h in the dark at 37 °C (98.6 °F)|
Ca – pantothenate – standard for the use of spiking food, feed and pharmaceutical products.
|Test Format||3 x Ca – pantothenate spiking standard (solid)|