疎水性相互作用あるいは親水性相互作用を利用する コアシェル型カラムHPLC法による 水溶性ビタミン類の分離分析疎水性相互作用あるいは親水性相互作用を利用する コアシェル型カラムHPLC法による 水溶性ビタミン類の分離分析AN00242368 Separation of Water Soluble Vitamins by HPLC with Core-shell Type Columns Utilizing Hydrophobic Interaction and/or Hydrophilic Interaction
This paper describes HPLC separation of water soluble vitamins such as ascorbic acid （vitamin C, VC） etc. employing the core-shell type column utilizing hydrophobic interaction and/or hydrophilic interaction. As for 12 water soluble vitamins, riboflavin （vitamin B2, VB2）,riboflavin sodium phosphate （vitamin B2 phosphate）, cyanocobalamin （vitamin B12）,hydroxocobalamin and mecobalamin were successfully separated by the reversed-phase （RP）HPLC with an ODS column employing 15 – 20 % acetonitrile（ ACN） or methanol（ MeOH） as an organic modifier. Other water soluble vitamins eluted at almost t0 （column hold up time）by the RP-HPLC with an ODS column using no addition of ACN or MeOH to the mobile phase.These seven water soluble vitamins （VC, isoascorbic acid, ascorbic acid 2-glucoside, nicotinic acid, nicotinamide, vitamin B1 and vitamin B6） were successfully separated by hydrophilic interaction chromatography（ HILIC） with an amide type column employing around 70% ACN mobile phase. The elution order in the HILIC mode was reversed compared with that in the RP-HPLC mode. Further each sample was identified more selectively by the UV spectra obtained by the photo diode array detector. Assay of pharmaceutical preparations and soft drink containing VC was successful by the HILIC mode HPLC employing an internal substance method within a short analysis time.