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《Modern Agricultural Sciences and Technology》 2010-07
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High Performance Liquid Chromatographic Analysis of 5-(3-Indolylmethyl) hydantoin and Its Products Converted by Engineered Bacteria

QIAN Kai XU Lin* YAN Ming (College of Biotechnology and Pharmaceutical Engineering,Nanjing University of Technology,Nanjing Jiangsu 210009)  
The HPLC method was used to determine the concentrations of substrate 5-(3-Indolylmethyl)hydantoin,intermediate product N-carbamyl-D-Tryptophan and product D-Tryptophan in the reaction solution. The sample was determined using a Purospher STAR RP-C18e column (4.6 mm×250.0 mm,5 μm particle size)at the temperature of 30 ℃ with the mobile phase of acetonitrile-20mmoL/L Sodium acetate(25∶75 by volume) which adjusted to pH 3.0 with phosphoric acid at a flow rate of 1.0 mL/min. The detection wavelength was set at 220 nm.Under these conditions,the components in the reaction system were separated completely within 10 min. A good linear relationship was obtained by employing external standard method for quantitative analysis. The linear calibration ranges for the three target analytes were 0.10~1.00 mg/mL(r=0.999 1),0.20~2.00 mg/mL(r=0.998 9)and 0.20~2.00 mg/mL(r=0.998 9),respectively. The recoveries of the analytes from spiking experiments were 99.3%~102.0% and 98.0%~ 99.3%,respectively,with the relative standard deviation being 0.51%~0.87% and 0.43%~0.94%,respectively. The results showed that the method was rapid,convenient and accurate.
【Fund】: 新一代生物催化和生物转化的科学基础(2009CB724700)
【CateGory Index】: Q814.9
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【Citations】
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【Co-citations】
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