RP-HPLC法测定发酵液中麦考酚酸的含量

RP-HPLC determination of mycophenolic acid in fermentation broth

目的:建立发酵液中麦考酚酸含量的反相高效液相色谱分析方法。方法:发酵液样品用甲醇提取后进样。采用 Sino-Chrom ODS-BP(5μm,200 mm×4.6 mm)色谱柱,以甲醇-10 mmol·L^(-1)醋酸铵(67:33)为流动相,流速1.0 mL·min^(-1),检测波长220 nm,艾瑞昔布为内标,测定了63株菌发酵液样品中麦考酚酸的含量。结果:发酵液中麦考酚酸含量在0.30～6.00mg·mL^(-1)范围内线性关系良好(r=0.9997,n=6),平均回收率为102.4%(n=9)。结论:本测定方法准确、简便,重复性好,为筛选发酵法合成麦考酚酸的高产菌株提供了可靠依据。

Objective： To develop an HPLC method for the determination of mycophenolic acid （MPA） in fermentation broth. Method： The fermentation broth was extracted by methanol, a SinoChrom ODS-BP column（5 μm,200 mm ×4.6 mm） with a solvent system consisting of methanol- 10 mmol · L^-1 ammonium acetate （67： 33） was used ,flow rate was 1.0 mL · min^-1 and the detection wavelength was 220 nm. Imrecoxib was used as the internal standard, and 63 batches of fermentation broth samples were studied. Results： The calibration curve was linear over the range of 0..30 -6. 00 mg · mL^-1 with the correlation 0. 9997 （n = 6）. The average recovery of mycophenolic acid was 102. 4% （n = 9 ）. Conclusion：The method is accurate, simple with good reproducibility, which is suitable for the determination of mycophenolic acid in fermentation broth.