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反相高效液相色谱法分离测定生孢乳杆菌发酵液中的乳酸

Determination of Lactic Acid in Fermentation Broth of Sporolactobacillus inulinus by Reversed-phase High Performance Liquid Chromatography (RP-HPLC)

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【作者】 付卫明白冬梅班睿赵学明李鑫钢徐世民

【Author】 Fu Weiming, Bai Dongmei, Ban Rui, Zhao Xueming, Li Xingang, Xu Shimin (Department of Biochemical Engineering, School of Chemical Engineering & Technology; National Engineering Research Center for Distillation Technology, Tianjin University, Tianjin, 300072)

【机构】 天津大学化工学院生物工程系天津大学精馏技术国家工程研究中心

【摘要】 采用反相高效液相色谱法测定了生孢乳杆菌发酵液中的乳酸。在Eclipse XDB-C8, (4.6 mm i.d.× 150 mm,5 μm)色谱柱上,以 0.005 mol/L硫酸(pH值2.5)溶液作流动相,流速为1 mL/min,以示差折光检测器检测信号,峰面积(Y)与乳酸的质量浓度(x,g/L)的线性相关方程为Y=292.08x+4.696 7,线性相关系数r为0.999 9,线性范围为0.75-9.0g/L,检测限为0.2μg(3倍信噪比)。结果表明,实验方法简便、快速、可靠,精密度为0.22%(m=5),回收率99%以上。

【Abstract】 A method for determining lactic acid in fermentation broth of Sporolactobacillus inulinus by RP -HPLC was described. The operating conditions were Eclipse XDB - C8 column (4.6 mm i. d. × 150 mm, 5 μm) at room temperature, 0. 005 mol/L sulfic acid solution (pH 2. 5) as mobile phase with flow rate of 1.0 mL/min and the detector is refractive index detector, the retention time of lactic acid is 2.243min. The linear correlation equation was Y = 292. 08x + 4. 696 7 ( r = 0. 999 9), the linear range is 0. 75 -9. 0g/L, and the detection limit is 0. 2 μg (signal to noise ratio is 3). This method is simple, quick, accurate and reproducible, the result is not affected by other components of fermentation broth. The relative standard deviation (RSD, n=5) was 0.22% , and the average recovery was above 99% .

【基金】 国家自然科学基金(No.20036010);博士点基金(No.2000005622)资助项目。
  • 【会议录名称】 第三届全国传质与分离工程学术会议论文集
  • 【会议名称】第三届全国传质与分离工程学术会议
  • 【会议时间】2002-11
  • 【会议地点】中国天津
  • 【分类号】TQ920.1
  • 【主办单位】中国化工学会
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