目的 建立纖溶酶反相高效液相色譜純度測定方法。方法 采用SHISEIDO CAPCELL PAK C₁₈ SG300（250 mm×4.6 mm，5 μm），以0.1%三氟乙酸溶液-乙腈（95：5）為流動相A，以0.08%三氟乙酸溶液-乙腈（20：80）為流動相B，進行梯度洗脫，體積流量為0.7 mL·min^-1，檢測波長280 nm，柱溫40℃。參照2020年版《中國藥典》四部9101藥品質(zhì)量標準分析方法驗證指導(dǎo)原則進行專屬性、檢測限、耐用性考察。應(yīng)用所建立的方法和分子排阻色譜法對3批樣品純度進行檢測。結(jié)果 建立的反相色譜法專屬性強，檢測限為0.01%，耐用性強。對3批樣品進行檢測，質(zhì)量分數(shù)在93.5%~96.9%，結(jié)果準確。結(jié)論 建立反相色譜法測定纖溶酶純度，分辨率更高、重現(xiàn)性更好，可進行更好的質(zhì)量控制。

Objective To establish a method of reversed-phase high performance liquid chromatography to determine the purity of fibrinolytic enzyme. Methods Using SHISEIDO CAPCELL PAK C₁₈ SG300 (250 mm×4.6 mm, 5 μm) or equivalent performance column. The mobile phase A was 0.1% trifluoroacetic acid solution - acetonitrile (95: 5), and the mobile phase B was 0.08% trifluoroacetic acid solution: acetonitrile (20:80) with gradient elution at the flow rate of 0.7 mL·min^-1, the detection wavelength was 280 nm, and the column temperature was 40 ℃. The specificity, detection limit and durability were investigated by referring to the guiding principles of drug quality standard analysis method validation of IV 9101 of Chinese Pharmacopoeia (2020 edition). The method and molecular exclusion chromatography were used to detect the purity of three batches of samples. Results The established reversed-phase chromatography has strong specificity, detection limit of 0.01% and durability. Three batches of samples were tested, the mass fraction was 93.5%—96.9%, and the results were accurate. Conclusion A reversed-phase chromatography method was established for the determination of plasminase purity with higher resolution, better reproducibility and better quality control.

R917