A reaction between a reducing sugar and 1-phenyl-3-methyl-5-pyrazolone (PMP) can produce a reducing sugar-PMP derivative with a high UV absorbance at 248 nm, which can be detected by high performance liquid chromatography (HPLC) coupled with UV detector or diode array detector (DAD). Response surface methodology (RSM) was applied to maximize the yields of the PMP derivatives of glucose, glucuronic acid, galacuronic acid and glucose-6-phasphate, which were incubated in water bath within a temperature range from 60°C to 90°C, and time range from 90 to 180 minutes. As a consequence, the optimal condition of glucose-PMP reaction was obtained at 71°C for 133 minutes, while the optimal conditions of glucuronic acid and galacuronic acid were obtained at 74°C for 151 minutes, and 76°C for 144 minutes, respectively. Additionally, the glucose-6-phasphate-PMP derivative had its optimal condition at 73°C for154 minutes. This study demonstrated that the PMP derivatives had significantly increased sensitivity and accuracy suitable for the HPLC analysis. In addition, the derivatization of different reducing sugars is subjected to their delicate functional groups, but sugar stereoisomers might have no remarkable effect on the yield and the optimal condition of the sugar-PMP reaction.

Optimization of reducing sugar-PMP reactions by response surface methodology

255th National Meeting (2018)

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