基于酰胺基柱的HPLC-RID同时测定异麦芽酮糖中5种糖的含量及其影响因素OACSTPCD

Simultaneously determination of the contents of fructose,glucose,sucrose,isomaltulose and trehalulose in isomaltulose products by High Performance Liquid Chromatography-Refractive Index Detector(HPLC-RID)and their influencing factors was studied.The method used an amide-based column,the accuracy of the method was confirmed by precision test and t-test,the effect of chromatographic conditions and pretreatment conditions on the separation was determined by univariate trials and spectral comparisons.The results showed that the increased proportion of acetonitrile in the mobile phase enhanced the retention of sugars on the amide-based column,during the separation process,triethylamine could significantly eliminate the split or bifurcation peaks produced by fructose,glucose and trehalulose,and affect the position of negative peaks in the spectrum;in the sample medium,acetonitrile mainly affected the polarity and size of the solvent peak,and triethylamine mainly affected the polarity and size of the negative peak.The volume fractions of acetonitrile and triethylamine in the mobile phase were 75％and 0.05％,respectively,column temperature was 45℃,flow rate was 1.0 mL/min,and injection volume was 10 μL,five sugars reached the baseline separation in 10 minutes and the column was safe for pH tolerance,there was no significant difference between the actual sample measurement results and COA(p=0.05),and the precision was less than 1.3％,which met the requirements of relevant testing standards.