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建立了对苯二甲酸、对甲基苯甲醛、对甲基苯甲醇、对甲基苯甲酸的高效液相色谱分析方法。采用HypersilSAX阴离子交换柱,流动相为2.5mol/LNH4H2PO4(含10%乙腈),pH4.32,流速0.8mL/min,柱温30℃,紫外检测波长为254nm。在此色谱条件下,各组分在7min内得到很好地分离,回收率符合测定要求。运用本方法测定了睾丸酮丛毛单胞菌和嗜麦芽窄食单胞菌生物催化生成对苯二甲酸不同发酵时间发酵液中主要代谢物含量。同时,采用GC-MS方法检测了有机酸、氨基酸、糖及长链脂肪酸等胞内代谢物,结合HPLC和GC-MS检测结果,分析了嗜麦芽窄食单胞菌和睾丸酮丛毛单胞菌协同作用催化对二甲苯生成对苯二甲酸的代谢途径。
A high performance liquid chromatographic method for the determination of p-toluic acid, p-methylbenzaldehyde, p-methylbenzyl alcohol and p-toluic acid was established. HypersilSAX anion exchange column was used. The mobile phase consisted of 2.5mol / L NH4H2PO4 (containing 10% acetonitrile), pH4.32, flow rate 0.8mL / min, column temperature 30 ℃ and UV detection wavelength 254nm. Under the chromatographic conditions, the components were well separated in 7min, the recovery rate in line with the determination requirements. The main metabolites in the fermentation broth of terephthalic acid at different fermentation time were determined by this method. At the same time, the intracellular metabolites such as organic acids, amino acids, sugars and long-chain fatty acids were detected by GC-MS. The results of HPLC and GC-MS analysis showed that Stenotrophomonas maltophilia and Comamonas testosteroni Metabolic pathway catalyzed by para-xylene to produce terephthalic acid.