Determination of Vitamin A,Vitamin D and Vitamin E in formulas for special medical purposes by online purification-two-dimensional liquid chromatography
Objective To determine vitamin A(VA),vitamin D(VD)and vitamin E(VE)in formulas for special medical purposes,the online purification-two-dimensional liquid chromatography technique with dual-valve triple-pump center-cutting was established.Methods After the saponification,the samples were introduced into a two-dimensional liquid chromatography system.A PLRP-S column,which was resistant to strong alkaline solutions,was used as an online solid phase extraction column for the clean-up of the sample solution.Subsequently,SPE pumps were reversed to elute the target to the one-dimensional chromatographic system.A perfluorophenyl column was used as the 1D column for the separation of VA and the four VE species.Meanwhile,a C18 column was used as a capture column for VD and a polycyclic aromatic hydrocarbon column was used as a two-dimensional chromatographic column for the separation of VD2 and VD3.Results VA,VD and VE exhibited good linearity within different concentration ranges.The linear ranges were 0.010 0-1.000,0.001 00-0.100,0.200-20.0 μg/mL,and the correlation coefficients were all as high as 0.999 9.The limits of detection(LODs)were 4.8,0.8 and 48 μg/100 g,and the limits of quantification(LOQs)were 12,2 and 120μg/100 g.The average recoveries of VA,VD and VE were good with the ranges of 85.9%-1 14.0%,and the relative standard deviations(RSDs)ranged from 0.6%to 9.2%for the different spiked levels of low,medium and high.Conclusion The method is easy to operate,with high sensitivity and good accuracy,and it is suitable for the routine rapid determination of VA,VD and VE in formulas for special medical purposes.
Online purificationtwo-dimensional liquid chromatographyvitamin Avitamin Dvitamin Eformulas for special medical purposes