A novel micellar thin layer chromatographic method for simultaneous separation and on-plate identification of B-group vitamins along with ascorbic and folic acids has been developed. A hybrid mobile phase constituting a mixture of 4% aqueous Sodium Dodecyl Sulphate (SDS) + acetonitrile (1:2, v/v) was identified as the most favorable for the resolution of multicomponent mixture of vitamins on silica high performance Thin Layer Chromatography (TLC) plates (silica gel 60F₂₅₄ catalog no. 1.05554, Merck, Germany). The resolved spots were identified by visualization under ultraviolet radiation (l = 254) in a closed UV cabinet. The effects of type of sample solvent, concentration of surfactant (SDS), volume ratio of acetonitrile in the mobile phase, and the presence of essential amino acids in the sample have been examined. This method is simple, precise, sensitive and useful for the analysis of vitamins B₁, B₂, B₆, B₁₂, ascorbic and folic acids present in marketed pharmaceutical formulation.

Thin Layer Chromatography (TLC) represents one of the most important analytical techniques for identification and simultaneous separation of organic compounds (Wagner and Bladt, 1995; Sherma and Fried, 2003; Cimpoiu and Hosu, 2007; and Mohammad and Laeeq, 2007). Vitamins, the amphipathic molecules, are the precursors of various metabolic pathways in every living organism. These organic molecules essentially play important functions as coenzymes and antioxidants. These occur in almost all dietary products. Their deficiency may lead to numerous disorders. To overcome their loss in pathological conditions, they can be used in the form of various therapeutic multivitamin dosages, which are composed of B-group vitamins, folic acid and ascorbic acids, biotin, calcium pantothenate fortified with minerals and antioxidants. For the analysis of six hydrophilic vitamins, this new micellar TLC procedure is designed.

According to literature, various methods like High Performance Liquid Chromatography (HPLC) using diode array detector in commercial multivitamin preparation (Chu K-On and Tin, 1998); HPLC (Huang X et al., 1999), Reverse Phase-HPLC (RP-HPCL) (Bhushan and Meenakshi, 2002), TLC using fiber-optic fluorometric quantitation (Navas et al., 1993) and quantitative determination by fluorescence quenching (Srinivas et al., 1992; and Perisic et al., 1995), High-Performance Thin Layer Chromatography (HPTLC) (Kartsova and Koroleva, 2007) and RP-TLC (Baranowska and Kadziolka, 1996) have been used in the analysis of vitamins. Most of thin layer chromatographic procedures developed for their analysis involve the use of organic or mixed aqueous organic mobile phases (Watanabe and Miyamoto, 2006). Since the first report (Armstrong and Terrill, 1979) on the use of aqueous solution surfactant as mobile phase, several workers (Dorsey et al., 1983) have utilized it in the analysis of closely related organic as well as inorganic substances by ion-pair and micellar TLC (Kord and Khaledi, 1992; Mohammad and Jabeen, 2003; and Mohammad et al., 2004).

Hydrophilic Therapeutic Vitamins, Micellar Thin Layer Chromatography, Micellar TLC, Anionic surfactant, Vitamin identification, Thin Layer Chromatography, TCL, Sodium Dodecyl Sulphate SDS, High Performance Liquid Chromatography, HPLC, High-Performance Thin Layer Chromatography, HPTCL.