1.0PRINCIPLE

Samples are prepared by mixing with an extraction solution(or just centrifuging), blending and filtering.The extract is then applied to the Vitamin B12 immunoaffinity column bound with specific antibodies to Vitamin B12．At this stage, the Vitamin B12 bind to the antibody on the column. The column is then washed with water to remove the impurities. By passing methanol through the column, the Vitamin B12 is removed from the antibody. This methanol solution can then be injected into HPLC or LC-MS/MS system.

2.0 INTENDED USE

A simple and efficient extraction and purification procedure for Vitamin B12 was developed by means of the immunoaffinity column (IAC - SEP ® VB12) as a cleanup tool. This method do not use toxic solvents such as chloroform and methylene chloride. Vitamin B12 content in Soft Drinks, Milk, Infant Formula, Food and Energy Bars are cleaned up by IAC and determined by HPLC or LC-MS/MS. It is a fast,simple, safe and highly accurate method for quantitatively measuring Vitamin B12.

The method can be used for the determination of vitamin B12 ,including Cyanocobalamin, the form used for fortification, and naturally occurring forms (Mecobalamin, Adenosine cobalamin and Hydroxocobalamin), which are predominantly found in meat and dairy products. To estimate the concentration of vitamin B12 in infant formulas and adult nutritionals containing not only added cyanocobalamin, but also natural forms, the vitamin B12 is freed from complexing proteins by heating the samples for a short time in the presence of cyanide. The use of an immunoaffinity extraction step increases selectivity and sensitivity by purifying and concentrating the food extracts. Vitamin B12 can then be determined by LC with UV detection at 361 nm or LC-MS/MS

3.0 APPLICATIONS

Milk ,Powdered Milk,Infant rice flour, Soft Drinks, Infant Formula,

Food and Energy Bars. etc.

4.0 ORDERING INFORMATION

Cat. No. Description

IAC 315 25 IAC/box

5.0 Specifications