Isocratic liquid chromatography technique for the analysis of cyanocobalamin from beef liver and heart muscle extracts

Reverse-phase high-performance liquid chromatography (RP-HPLC) is among the most widely recommended techniques for analyzing water-soluble vitamins such as vitamins C and B-complexes. The research study was conducted to detect and quantify cyanocobalamin (vitamin B12) from the beef liver and heart muscle extracts using a validated isocratic RP-HPLC procedure. The analytical column was Luna® Phenomenex 5 µm C18 (2) 100 A LC-column (150 × 4.6 mm). The mobile phase consisted of water/ethanol in a ratio of 60:40 (v/v). An enzymatic digestion with 1% potassium cyanide was used for samples (beef liver and heart muscle) extraction. The validated method showed to be linear, R = 0.9977; fast, with a retention time of less than 6.00 min; precise, %RSD of 1.16 and 1.74%; and sensitive, with limits of detection and quantification of 0.00033 and 0.00100 µg/mL. The detected values of cyanocobalamin from the beef liver (BLV) and heart muscle (HRM) extracts were 52.04 ± 0.13 and 42.04 ± 0.29 µg/mL, respectively. BLV extract indicated a higher level of cyanocobalamin. Hence, the validated isocratic RP-HPLC technique can be recommended to analyze cyanocobalamin and related compounds in research laboratories such as diagnostics, foods, and pharmaceuticals.