The measuring range is linear of 25ng to 1250ng Vitamin B12 pro injection (R2=0.9999). The limit of detection is 3ng of vitamin B12 per injection (three times of signal/noise ratio). If the given dilution steps are followed, the vitamin B12 contents of 0.1 to 5μg/g lie within the linear working range of the method. If the contents of the samples used are higher than the cited range, extracts should be diluted in a suitable manner. The lower limit of detection is 10ng/g of vitamin B12 in the sample. Range: 0.01 - 5μg. Sensitivity: 0.01 μg. ? Many methods of Vitamin B12 determination are based on HPLC-UV (High Performance Liquid Chromatography-ultraviolet) detection show low selectivity if problematic matrices are applied. This method of content determination of Vitamin B12 combines the high selectivity of immunoaffinity columns with its potential to concentrate, elute, and purify with the HPLC column.