Simultaneous Determination of B₁, B₂, B₃, B₆, B_9, and B₁₂ Vitamins in Premix and Fortified Flour Using HPLC/DAD: Effect of Detection Wavelength

Table 3

System suitability test results for the HPLC method for determination of B vitamins (9 injections of aqueous standard solution containing 5 mg/L).

Vitamin	Wavelength (nm)	RT (min)^a	% RSD of RT	PA	% RSD of PA	Capacity factor (K)	Selectivity (α)	Resolution (RS)	Tailing factor (TF)	TP
B1	265	4.37	1.40	2.65	0.79	0.73	1.32	3.41	1.23	12020
	280	4.32	1.45	1.13	1.55	0.72	1.33	3.45	1.23	11896
B3	265	4.96	1.31	2.49	0.63	0.97	1.54	9.96	1.41	10815
	280	4.92	1.52	0.24	2.30	0.95	1.57	10.72	1.29	10940
B6	265	6.29	0.39	0.67	2.43	1.50	1.46	21.65	1.85	117192
	280	6.29	0.39	2.16	1.37	1.50	1.46	22.15	1.21	130915
B9	265	8.02	0.24	1.42	1.38	2.18	1.05	3.08	1.15	137022
	280	8.02	0.24	2.1	0.92	2.18	1.05	3.04	1.15	143606
	361	8.02	0.23	0.46	2.44	2.18	1.05	3.06	1.16	142637
B12	265	8.30	0.18	0.94	0.74	2.29	1.16	8.05	1.11	103467
	280	8.30	0.18	1.06	0.89	2.29	1.16	7.94	1.12	99293
	361	8.30	0.21	1.91	1.45	2.29	1.16	8.03	1.11	107767
B2	265	9.23	0.37	5.47	1.33	2.66	—	—	1.12	78402
	280	9.23	0.37	3.65	2.35	2.66	—	—	1.12	80602
	361	9.23	1.37	1.68	2.54	2.65	—	—	1.11	80869

^a The retention time of unretained peak is 2.52 min. RT: retention time, RSD: relative standard deviation, PA: peak area, and TP: theoretical plates.