Vitamin B12 (Cyanocobalamin)
Novel immunoaffinity columns with built-in reservoir for detection of vitamin B12 in a wide range of commodities, developed in conjunction with Nestle Research Centre, Switzerland.
The procedure is based on monoclonal antibody technology, which makes the test highly specific, sensitive, rapid and simple to perform. Improved clean-up and concentration of toxins from complex food matrices results in reduced chromatography interference and lower detection limits.
|Art. No.||P88 / P88B|
|Test Format||10 large reservoir format immunoaffinity columns (P88).
50 large reservoir format immunoaffinity columns (P88B).
|Sample Preparation||The columns contain a gel suspension of monoclonal antibody specific to the vitamin of interest. The immunoaffinity columns are designed with an integrated reservoir and a tightly fitting top and bottom cap. The top frit is absent from the top of the antibody gel to enable more thorough mixing of the sample extract and the antibody within the column. Following extraction of the vitamin the sample extract is filtered and the filtrate is added to the reservoir. The column is then placed on a rotary shaker where the sample mixes directly with with the gel allowing binding to take place between the antibody and the vitamin. The column is then washed to remove any unbound material and the vitamin is released following elution with solvent. The eluate is collected, evaporated and reconstituted prior to analysis by HPLC or LC-MS/MS. The extra contact between the extract and the antibody results in a more uniform binding of the vitamin with the antibody and leads to better repeatability and reproducibility.|
|Detection Limit||LOD: 5 ng/ml.|
|Shelf life||18 months at 2 – 8 °C (or 12 months at 21 – 25 °C).|
|Sensitivity||Low % CV, excellent recoveries and detection limit.|
|Approvals||Columns have been validated and approved by Nestlé. Received AOAC Official First Action Method 2011.08.|
|Rapid||Clean up 2 hours prior to detection by HPLC or LC-MS/MS.|
|Robust||Can be stored at room temperature.|
|Reliable||Uses a monoclonal antibody to selectively isolate and concentrate vitamin B12.|
|Adaptable||Suitable for analysis of fortified or total vitamin B12. Methods are available for a wide range of commodities.|
|Extraction solvents||Distilled water or sodium acetate buffer (pH 4).|
RIDASCREEN®FAST Vitamin B12
RIDASCREEN®FAST Vitamin B12 is a competitive enzyme immunoassay for the quantitative analysis of vitamin B12 in fortified food, feed and vitamin products.
Vitamin B12 is one of the essential coenzymes in the vertebrate metabolism. The most important functions of vitamin B12 are blood cell synthesis, fatty acid metabolism and the availability of folic acid. Folic acid itself plays a considerable role in the biosynthesis of nucleotides as well as in the turnover of amino acids. A deficiency of vitamin B12 may cause serious disorders such as pernicious anemia or neurological symptoms.
|Test Format||Microtiter plate with 48 wells (6 strips with 8 removable wells each)|
|Standard Range||0 ppb (zero standard), 5 ppb, 10 ppb, 30 ppb, 100 ppb, 300 ppb|
|Sample Preparation||milk: centrifugation and dilution
juices: dilution, filtration or centrifugation
other matrices: homogenization, extraction, filtration or centrifugation and dilution
|Incubation Time||25 min|
|Detection Limit||raw milk: approx. 0.5 ppb
pasteurized milk: approx. 2.5 ppb
milk mix drinks: approx. 2.5 ppb
milk powder: approx. 10 ppb
grain, flour, cereals: approx. 25 ppb
vitamin powder, -premixes, -tablets, -capsules and -candies: approx. 25 ppb
vitamin juice: approx. 2.5 ppb
|Cross Reactivity||Vitamin B12: 100 %
Vitamin B12a (Hydroxycobalamin): approx. 80 %
VitaFast® Vitamin B12 (Cyanocobalamin) (AOAC-RI)
Easy to use microbiological microtiter plate test for the quantification of total vitamin B12 (enriched and natural vitamin B12) in food, feed and pharmaceutical products.
Vitamin B12 is extracted from the sample and the extract is diluted. The diluted extract and the vitamin B12 assay-medium are pipetted into the wells of a microtiter plate which is coated with Lactobacillus delbrueckii subsp. lactis (leichmannii). The growth of Lactobacillus delbrueckii is dependent on the supply of vitamin B12. Following the addition of vitamin B12 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 vitamin B12 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||0.03 – 0.18 µg / 100 g (ml)|
|Sample Preparation||For the determination of added vitamin B12 in nutrient enriched solid samples, a hot water extraction is usually sufficient. Liquid samples can be used directly after sterile filtration. For the determination of the total vitamin B12 (native and added) content, the sample has to be treated with NACN/KCN and enzyme.|
|Incubation Time||44 – 48 h in the dark at 37 °C (98.6 °F)|
VitaFast® Vitamin B12 (Cyanocobalamin) Dotierstandard/Spiking Standard
Vitamin B12 standard for the use of spiking food, feed and pharmaceutical products.
|Test Format||3 x vitamin B12 (cyanocobalamin) spiking standard (solid)|