菱形藻小分子有机酸的提取及HPLC测定

Extraction of organic acid with low molecular weight and determination by HPLC in Nitzschia sp.

采用Agilent ODS C18(φ5μm,4.6mm×250mm)柱,以0.5%(NH4)2HPO4-H3PO4为流动相(pH=2.4),检测波长为214nm,流速为1.0mL/min,在12min内能对草酸、酒石酸、甲酸、丙酮酸、苹果酸、维生素C、乙酸、马来酸、柠檬酸、琥珀酸和富马酸等11种小分子有机酸进行有效分离,各组分线性关系良好,相关系数均在0.9991以上。采用2%偏磷酸做提取剂,超声波辅助提取,可有效提取菱形藻(Nitzschiasp.)胞内小分子有机酸,细胞中检测到除琥珀酸外的10种小分子有机酸;培养基中检测到除柠檬酸和琥珀酸外的9种小分子有机酸。

The conditions of simultaneous determinations of oxalic,tartaric,formic, pyruvie, malic, ascorbic,acetic, maleie ,citric ,sueeinic ,fumaric acid by reversed-phase HPLC in 12 min were studied ,using Agilent ODS C18 （φ5 μm,4.6 mm×250 mm）-column,with 0.5 % （NHa）2HPO4-H3P04 （pH=2.4）as mobile phase, by UV detector at 214 nm,with flow rate of 1.0 mL/min.The linearity of each component was excellent and all the correlative coefficients exceeded 0.9991.Using 2 % metaphosphorie acid as extractant could extract the molecular weight organic acids of Nitzschia sp. effectively, with ultrasonic-assisted extraction.Ten kinds of organic acids except of succinic acid were detected in cell,and nine kinds organic acids except of citric and succinic acid were detected in culture medium.