Isolation and quantitative analysis of B1, B2, B6 AND B12 vitamins using high-performance thin-layer chromatography

Abstract

A new, simple and accurate high-performance thin-layer chromatography (HPTLC) method has been developed for the separation and simultaneous analysis of B1, B2, B6 and B12 vitamins. Standard and sample solutions of vitamins were applied onto precoated aluminum-backed silica gel G 60 F254 HPTLC plate. The plates were developed with about thirty different solvent systems, and the ethanol -chloroform -acetonitrile -toluene -ammonia -water (7 : 4 : 4.5 : 0.5 : 1 : 1, v/v) mixture was chosen as the optimum mobile phase. UV detection was performed densitometrically at 254 nm. The retention factors of B1, B2, B6 and B12 vitamins were 0.36, 0.60, 0.85 and 0.46, respectively. The linearity range for indicated compounds was 10 -2500 ng per spot and the correlation coefficients were R 2 = 0.97 -0.98. The limits of quantification (LOQ) were 141.72, 42.41, 100.31 and 11.50 ng and the limits of detection (LOD) were 42.52, 12.72, 30.09 and 3.45 ng for B1, B2, B6 and B12 vitamins, respectively. Analyses of standard solutions showed good precision and repeatability.

A Review of the Extraction and Determination Methods of Thirteen Essential Vitamins to the Human Body: An Update from 2010

“…Besides the methods mentioned above, several effective methods have also been developed, e.g., immunoassays [ 99 ], high performance thin-layer chromatography [ 81 ] and supercritical fluid chromatography [ 98 ]. The use of these methods is justified when it is necessary to carry out routine quality control of relatively simple sample compositions.…”

“…Recently, TLC has been improved to incorporate HPTLC grade stationary phases, automated sample application devices, a controlled development environment, automated development, forced-flow techniques, computer-controlled densitometry, quantitation and fully validated procedures. HPTLC is becoming a routine analytical technique because of its advantages of low operating cost, high sample throughput, simplicity, speed, need for minimum sample clean up, high reproducibility, accuracy, reliability and robustness [ 81 ]. Panahi et al [ 81 ] isolated and quantified vitamins B 1 , B 2 , B 6 and B 12 using HPTLC.…”

“…HPTLC is becoming a routine analytical technique because of its advantages of low operating cost, high sample throughput, simplicity, speed, need for minimum sample clean up, high reproducibility, accuracy, reliability and robustness [ 81 ]. Panahi et al [ 81 ] isolated and quantified vitamins B 1 , B 2 , B 6 and B 12 using HPTLC. The precoated aluminum-backed silica gel G60 F254 HPTLC plate was developed with nearly 30 different solvent systems.…”