Document Detail

Characterisation of vitamin B12 immunoaffinity columns and method development for determination of vitamin B12 in a range of foods, juices and pharmaceutical products using immunoaffinity clean-up and high performance liquid chromatography with UV detection.
MedLine Citation:
PMID:  19680900     Owner:  NLM     Status:  In-Process    
New rapid and simpler procedures, using immunoaffinity columns, have been developed for the determination of vitamin B12 in a range of samples including three different US National Institute of Standard and Technology (NIST) Reference Materials, infant formula, powdered energy drinks and bars, wheat breakfast cereal, carbonated soft drinks, fruit juices and vitamin B12 tablets. The procedures involved extraction of vitamin B12 using water or sodium acetate buffer and enzyme digestion (using pepsin or alpha-amylase, or both) if necessary. The extract was clarified and passed through "EASI-EXTRACT Vitamin B12", an immunoaffinity column containing monoclonal antibody with high affinity and specificity to vitamin B12. Subsequently, the vitamin B12 immunoaffinity column was washed with 10 ml water and the vitamin B12 was released from the column with 3 ml methanol. Following evaporation, the samples were reconstituted in mobile phase and analysed by HPLC-UV at 361 nm on an ACE 3AQ analytical column using a gradient elution consisting of 0.025% trifluoroacetic acid in water and acetonitrile. Analysis of three types of NIST Standard Reference Materials in triplicate demonstrated the results of the immunoaffinity column method were comparable to microbiological assay results. Method repeatability was determined for all samples analysed and ranged between 0.8 and 10%, demonstrating the method was repeatable with complex matrices (NIST 2383) containing low levels of vitamin B12 (0.44 microg per 100 g), as well as simpler matrices, such as vitamin tablets containing high levels (2000 microg per 0.849 g) of vitamin B12.
E C Marley; E Mackay; G Young
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment     Volume:  26     ISSN:  1944-0057     ISO Abbreviation:  Food Addit Contam Part A Chem Anal Control Expo Risk Assess     Publication Date:  2009 Mar 
Date Detail:
Created Date:  2009-08-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101485040     Medline TA:  Food Addit Contam Part A Chem Anal Control Expo Risk Assess     Country:  England    
Other Details:
Languages:  eng     Pagination:  282-8     Citation Subset:  IM    
Research Department, R-Biopharm Rhone (RBR) Limited, Glasgow G200SP, UK.
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