摘 要：目的 通過建立HPLC分析法測定血必凈注射液中氨基酸，為提高中藥注射液質(zhì)量標準提供依據(jù)。方法 通過 OPA-FMOC柱前衍生化法，建立同時檢測血必凈注射液中18種常見氨基酸的液相色譜法。色譜柱為Agilent SB-C18柱（250 mm×4.6 mm，5 μm）；流動相A為0.02 mol/L醋酸鈉溶液（含200 μL/L三乙胺和3.5 mL/L四氫呋喃，2%醋酸溶液調(diào)pH 7.2±0.05），流動相B為0.02 mol/L醋酸鈉溶液–甲醇–乙腈（200∶400∶400）（pH 7.2±0.05），梯度洗脫；體積流量：0～38 min為1.0 mL/min；柱溫40 ℃；檢測波長0～32.5 min為338 nm，32.5 min以后為262 nm。結(jié)果 所測18種氨基酸在各自的高、低質(zhì)量濃度范圍內(nèi)線性關(guān)系良好；3個批次血必凈注射液中，游離氨基酸測定平均值為1.320 mg/mL，總氨基酸測定平均值為2.027 mg/mL。結(jié)論 所建立的氨基酸定量方法結(jié)果準確可靠，重現(xiàn)性好，有助于血必凈注射液的質(zhì)量控制，對中藥注射劑安全評價標準的提高具有指導意義。

Abstract: Objective To enhance the standards of injection of Chinese materia medica (CMM) by establishing a RP-HPLC method for simultaneous quantitative determination of amino acids in Xuebijing Injection．Methods Liquid chromatographic fingerprint method for simultaneous determination of 18 kinds of common amino acids was established by o-phthalaldenyde-9-fluorenylmethyl chloroformate (OPA-FMOC) pre-column derivatization. The column was Agilent SB-C18 (250 mm × 4.6 mm, 5 μm), mobile phase consisted of 0.02 mol/L sodium acetate (A),including 200 μL/L triethylamine and 3.5 mL/L tetrahydrofuran with pH value (7.2±0.05) regulated by 2% acetic acid solution, and 0.02 mol/L sodium acetate- methanol-acetonitrile (200∶400∶400) with pH value (7.2 ± 0.05)(B)for gradient elution at the flow rate of 1.0 mL/min within 0―38 min. The detection wavelength was 338 nm within 0－32.5 min and 262 nm,after 32.5 min, while the column temperature was 40 ℃． Results There wsa a good linear relationship of 18 kinds of amino acids with a range of 0.001 2—0.195 4 mg/mL in both high and low concentration. In three batches of Xuebijing Injections, the mean value was 1.320 mg/mL for free amino acids and 2.027 mg/mL for total amino acids. Conclusion The optimized method is accurate and reproducible, and is suitable for the determination of amino acids in Xuebijing Injection. It contributes to the quality control of the injection and guides the improvement of safety evaluation standard of CMM injections.