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 3

Method optimisation results. Results from the sample preparation method optimisation for vitamins spiked in artificial plasma samples processed using each of the three methods mentioned above. Results are reported as mean of peak area ± RSD ( 𝑛 = 3 ). Numbers in parenthesis indicate wavelength used for quantitation.

Vitamins (nm) Peak area [mAU] % RSD

200 μL pure acetonitrile + solid phase extraction
 C (230) 150.92 2.6
 B1 (270) 62.60 1.7
 B2 (265)366.1610.3
 B5 (266)27.140.4
 B6 (257)110.282.4
 B9 (280)69.132.0
 B12 (230)179.135.2
L/L extraction_aqueous phase
 C (230)197.914.7
 B1 (270)64.102.9
 B2 (265)216.316.0
 B5 (266)14.830.2
 B6 (257)145.079.4
 B9 (280)112.930.9
 B12 (230)132.362.6
600 μL ethanol : methanol (95 : 5) + solid-phase extraction
 C (230)169.142.8
 B1 (270)87.752.3
 B2 (265)302.291.9
 B5 (266)45.520.7
 B6 (257)60.971.0
 B9 (280)285.335.4
 B12 (230)111.741.0