B-590.52.530 [2020-10] Pantothenic Acid – Microtiter Plate Test

Numerous beverages, such as multivitamin juice and sports drinks, are enriched with vitamins. These beverages may not be listed on a label as "enriched" or "fortified" with a particular vitamin supplement unless the vitamin is present in "significant amounts", which are defined in Annex XIII to Regulation (EU) No 1169/2011. For beverages, this amount is set at a minimum of 7.5 % of the NRV (nutrient reference values) per 100 ml or 15 % of the NRV per portion for individual portions.

Quantities recommended for the daily dietary intake of vitamins:

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Application/Purpose

Analysis for determining the amount of added pantothenic acid by means of the microbiological microtiter plate test

Scope of Application

This analysis is suitable for NAB, juice and other foods.

Principle

The VitaFast® microbiological assay for pantothenic acid is carried out on microtiter plates and is suitable for the determination of total amount of pantothenic acid in foods and beverages.

The beverage is sterile-filtered and diluted with sterile water for the determination of added pantothenic acid.

The sample must be digested through enzymatic hydrolysis in order to measure the total content of pantothenic acid (naturally occurring and added). 

The diluted sample and the pantothenic acid assay medium are added to the wells of the microtiter plate, which are coated with Lactobacillus plantarum. Lactobacillus plantarum requires pantothenic acid to grow. After an addition of pantothenic acid, the microorganism exhibits growth until the vitamin is completely utilized. The plates are incubated at 37 °C for 20–24 h.

The growth of Lactobacillus plantarum is dependent upon the concentration of pantothenic acid and is determined through measuring the turbidity. These results are compared with those obtained from a series of standard concentrations. The measurement is performed with a microtiter plate photometer at 610–630 nm (alternatively at 540–550 nm).

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