PERBANDINGAN ANALISIS KROMATOGRAFI CAIR KINERJA TINGGI ELUASI GRADIEN DENGAN ISOKRATIK PADA PENENTUAN VITAMIN B1, B2 DAN B6 DALAM SEDIAAN SIRUP MULTIVITAMIN SECARA SIMULTAN

The aim of this research is to compare the simultaneous analytical results of B1 (Tiamin), B2 (Riboflavin), and B6 (Piridoxin) in multivitamin syrup between gradient and isocratic methods using high performance liquid chromatography (HPLC). The separation was performed on 15 cm length of C18 column and a mixture of methanol, water and glacial acetate acid with ratio of 10:90:1 was used as solvent. The eluent was a mixture of methanol and 5 mM sodium hexane sulfonate in 0.5% glacial acetate acid and gradually mixed using 2 different pumps. The optimal analytical conditions for gradient method were found to be 2,0 mL/min of flow rate and 280 nm of wavelength. On this gradient method, the vitamins were completely separated with retention times of 3.441 minutes, 4.985 minutes, and 7.393 minutes for piridoxin, riboflavin and tiamin respectively. Meanwhile, isocratic method (mixture of 5 mM sodium hexane sulfonate with metanol of 70 : 30) on the same conditions can only separate riboflavin and piridoxin. The RSD values for riboflavin and piridoxin were 1.377% and 1.376% respectively on isocratic methods and 0.693% and 0.825% respectively on gradient method. The linearity of both methods have fulfilled the requirement of R2 values that are higher than 0.999 for all vitamins.