The Scientific World Journal

Research Article

A Novel HPLC Method for the Concurrent Analysis and Quantitation of Seven Water-Soluble Vitamins in Biological Fluids (Plasma and Urine): A Validation Study and Application

Table 2

System suitability test results for the optimised HPLC method for determination of water-soluble vitamins in biological matrices (column temperature = 30°C; flow rate = 0.2 mL min⁻¹). Mean peak areas of 5 injections of aqueous standard solution containing 20 ng mL⁻¹ of each water-soluble vitamin.


Vitamin	Retention time (min)^a (%CV)^b	Capacity factor (K′)	Selectivity (α)	Resolution ()	Tailing factor	plate count	Mean peak areas (mAU)^c (%CV)^b

C	3.5 (0.9)	1.72	1.20	2.88	0.79	3111	365 (0.01)
B₁	4.3 (0.7)	2.21	1.15	2.65	0.77	4943	170 (0.03)
B₃	5.3 (1.1)	2.79	1.17	3.13	0.80	6056	93 (0.02)
B₆ (PLP)	6.1 (0.99)	3.86	1.83	1.89	0.56	4255	879 (0.001)
B₅	6.8 (0.29)	3.36	1.15	2.54	0.85	7050	60 (0.05)
B₉	13 (0.07)	8.29	1.03	1.60	0.46	6735	205 (0.01)
B₁₂	13.4 (0.6)	8.57	1.03	1.27	0.42	2329	740 (0.007)
B₂	13.8 (0.2)	8.86	1.07	1.30	0.46	1096	2140 (0.001)

^aThe retention time of unretained peak is 1.4 min.
^b%Coefficient of variation.
^cmilli Arbitrary Unit.