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目的建立发酵液中他克莫司快速筛查检测方法。方法利用发酵提取物,通过筛选薄层板、展开剂和显色方法建立适于发酵液中他克莫司检测的薄层色谱法。利用薄层色谱法对10批次的发酵样品进行检测,同时用高效液相色谱法对其检测结果进行验证。结果建立了薄层色谱法,以丙酮∶正己烷∶甲酸=4∶6∶0.1为展开剂,在硅胶GF254板上展开,经5%的硫酸乙醇显色后在120℃下烤板,斑点清晰。其检测结果和高效液相色谱结果相一致。结论该方法简便经济,可用于菌种选育的快速筛查。
Objective To establish a rapid screening assay for tacrolimus in fermentation broth. Methods Thin layer chromatography (TLC) was developed for the determination of tacrolimus in the fermentation broth by using the fermentation extract by screening the thin plate, developing solvent and colorimetric method. Ten batches of fermented samples were detected by thin-layer chromatography, and their test results were validated by high-performance liquid chromatography. Results Thin layer chromatography (TLC) was developed on a silica gel GF254 plate with acetone: n-hexane: formic acid = 4: 6: 0.1 as developing solvent. The plate was baked at 120 ℃ with 5% sulfuric acid. . The test results and high performance liquid chromatography consistent. Conclusion The method is simple and economical and can be used for rapid screening of strain selection.