RP-HPLC同时检测 Zymomonas mobilis 代谢组分方法的研究

优化了高效液相色谱(HPLC)同时检测Zymomonas mobilis发酵液中葡萄糖、乙醇和多种代谢产物的条件和方法,分析了影响因素、回收率和重现性.试验结果表明,选择10 mmol/L pH值为2.5 KH2PO4-H3PO4溶液作为流动相、SunFireTM C18,5μm色谱柱(4.6 mm×150 mm)、二极管阵列检测器,可同时检测出葡萄糖酸、丙酮酸、乳酸、乙酸;选用2 mmol/L硫酸溶液作为流动相、IC-PakTM lon-Exclusion 50 A7μm色谱柱(7.8 mm×300 mm)、示差折光检测器,可同时检测葡萄糖、山梨醇、琥珀酸、乙酸、乙醇.Z.mobilis发酵液的回收率和重现性的结果表明,该方法测定葡萄糖、乙醇和其它产物的相对标准偏差范围为0.005%～0.018%,平均回收率为96.86%～101.99%.

SIMULTANEOUS DETERMINATION OF METABOLIZED COMPOSITION IN ZYMOMONAS MOBILIS FERMENTED LIQUID BY RP-HPLC

A new method was described for simultaneous determination of Z.mobilis metabolites by reversed phase-high performance liquid chromatography(RP-HPLC).The samples were analyzed on a C18 column(Waters SunFireTM C18,Φ 416 mm×150 mm,5 μm),and the mobile phase was 10 mmol/L pH2.5 KH2PO4 solution with a flow rate of 0.8 mL/min.Under this optimized condition gluconate,pyruvate,lactic acid and acetic acid were completely separated and determined with a photodiode array detector(PAD).The IC-PakTM on-Exclusion column(50 A,7 μm,7.8 mm×300 mm) were used and the mobile phase was 10 mmol/L sulfuric acid solution with flow rate of 0.6 mL/min.Glucose,sorbitol,succinic acid,acetic acid and ethanol were simultaneously determined by refractive index detector(RID).The standard deviation values and the average recovery rates were in the range of 0.005%-0.018%,96.86%-101.99%,respectively.