RP-HPLC determination of vitamins, folic acid and B12 in multivitamin tablets

Abstract

A simple and sensitive reversed-phase, ion-pair HPLC method was developed and validated for the simultaneous determination of B-group vitamins, thiamine chloride hydrochloride (B1), nicotinamide (B3), pyridoxine hydrochloride (B6) and folic acid in Pentovit® coated tablets. The cyanocobalamine (B12) was determined separately, because of its low concentration in the investigated multivitamin preparation. RP-HPLC analysis was per- formed with a LKB 2150 HPLC system, equipped with a UV/VIS Waters M 484 detector. The procedures for the determination of B1, B2, B6 and folic acid were carried out on a Supelcosil ABZ+ (15 cm 4.6 mm; 5 m) column with methanol-5mM heptanesulphonic acid sodium salt 0.1 % triethylamine TEA (25:75 V/V); pH 2.8 as the mobile phase. For the determination of B12 a Suplex pKb-100 (15 cm 4.6 mm; 5 m) column and methanol–water (22:78 V/V) as the mobile phase were used. The column effluents were monitored at 290 nm for B1, B3, B6 and folic acid, and at 550 nm for B12. The obtained results and statistical parameters for all the investigated vitamins of the B-group in Pentovit® coated tablets were satisfactory and ranged from 90.4 % to 108.5 % (RSD. from 0.5 % to 4.1 %). The parameters for the validation of the methods are given.

Za istovremeno određivanje vitamina B-kompleksa, tiamin-hlorida-hidrohlorida (B1), nikotinamida (B3), piridoksin-hidrohlorida (B6) i folne kiseline u Pentovit® tabletama primenjena je i validirana jednostavna i osetqiva metoda reverzno-fazne jon-par tečne hromatografije pod visokim pritiskom. Zbog niske koncentracije u ispitivanom preparatu cijanokobalamin (B12) je određen pod drugim uslovima. Određivanje vitamina B1, B3, B6 i folne kiseline izvedeno je na LKB 2150 HPLC sistemu, sa UV/VIS Waters M 484 detektorom, korišćewem Supelcosil ABZ+ kolone (15 cm x 4,6 mm; 5 _m) sa mobilnom fazom metanol – 5 mM natrijum-heptansulfonat/0,1 % trietilamin (25:75 V/V) pri pH 2,8. Za određivanje vitamina B12 korišćena je kolona Suplex pKb-100 (15 cm x 4,6 mm; 5 nm) i mobilna faza metanol–voda (22:78 V/V). Vitamini B1, B3, B6 i folna kiselina detektovani su na 290 nm, a vitamin B12 na 550nm. Dobijene vrednosti sadržaja i statistički parametri za sve vitamine B-kompleksa u Pentovit® tabletama su zadovoljavajući i kreću se u opsegu od 90,4%do 108,5% (RSD od 0,5%do 4,1 %). Prikazani su i parametri za validaciju metode.