Determination of vitamin D3 in vitamin D drops by high resolution sampling two-dimensional chromatography
Objective:To establish an innovative analytical method based on high resolution sampling two-di-mensional chromatography(HiRes 2D-LC)for determination of the content of vitamin D3 in vitamin D drops.Methods:Two-dimensional liquid chromatography was used.Thermo HYPERSIL Gold Silica(100 mm ×2.1 mm,1.9 µm)column was used in the first dimension with n-hexane-n-amyl alcohol(996:4)as mobile phase.The flow rate was 0.2 mL·min-1.The samples were injected and tested at the wavelength of 265 nm.The column temperature was 40 ℃.In the second dimension liquid chromatography,ShimPack Velox Hilic(50 mm × 2.1 mm,2.7 μm)was used as the column with n-hexane-n-pentanol-isopropanol(98∶1∶1)as the mobile phase.The flow rate was 0.5 mL·min-1.The samples were injected and tested at the wavelength of 265 nm.The column temperature was 40 ℃.A six-position 14-way valve and was equipped with 2 multi-center cutting valves was equipped to make multiple consecutive cuts of the pre-vitamin D3 peak and vitamin D3 peak.Results:The calibration curves showed a good linearity at the range of 1.018 4-5.092 mg·mL-1(r≥0.999 8).The precision test showed the RSDs of the peak area of pre-vitamin D3 and vitamin D3 were 0.95％and 0.40％,respectively.The repeatability test showed the RSD of vitamin D3 content was 0.41％.The average recovery rate(n=9)was 101.4％.The test solution was stable at 4 ℃ and 10 ℃ for 12 h,and the RSDs were 0.58％and 0.66％,respectively.The contents of vitamin D3 in the samples of vitamin D drops measured by this method were 100.4％,101.6％,100.9％,101.6％,102.7％and 101.6％,which was basically consistent with the results measured by the fourth method in General Chapter 0722 of ChP 2020 Vol Ⅳ.Conclusion:This method has a good specificity and high sensitivity to accurately determine the content of vitamin D3 in vitamin D drops.
high resolution samplingtwo-dimensional chromatographynormal plase chromatographyvitamin D dropspre-vitamin D3vitamin D3normal-phase chromatographypeak purity confirmation
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