目的：采用9-芴甲氧羰酰氯（FMOC-Cl）为柱前衍生化试剂，建立FMOC-Cl柱前衍生化反相高效液相色谱法测定L-鸟氨酸盐酸盐的有关物质。方法：采用Agilent Extend C18（250 mm×4.6 mm，5μm）色谱柱，醋酸盐缓冲溶液（0.1 mol·L-1醋酸铵，冰醋酸调节pH至5.0）-乙腈为流动相，梯度程序洗脱，流速：1 mL·min-1，检测波长：265 nm，柱温：35℃，测定L-鸟氨酸盐酸盐中的有关物质。衍生化方法与条件：需衍生化样品与0.05 mol·L-1硼砂缓冲液和8 mg·mL-1 FMOC-Cl乙腈溶液（v/v，1∶3∶3）室温反应20 min后正己烷萃取2次，4 mol·L-1盐酸溶液调节pH至3~4。结果：该品及其潜在的12种氨基酸类有关物质衍生物色谱分离度良好；测得各氨基酸浓度在1.0~20μg·mL-1范围内，线性关系良好，r均大于0.9995，加样回收率和精密度结果均良好。结论：该方法专属性好、准确度高，适用于该品有关物质的检测。%Objective: To establi sh a reversed-pha se high performance liquid chromatographic method with 9-fluorene methoxy carbonyl chloride (FMOC-Cl) pre-column derivatization for the determination of L-ornithine monohydrochloride related substances. Methods: The analysis was carried out on an Agilent Extend C18 column (250 mm×4.6 mm, 5μm) by using 0.1 mol·L-1 ammonium acetate (adjust to pH 5.0 with glacial acetic acid) and acetonitrile as mobile phases with a linear gradient elution mode at a flow rate of 1 mL· min-1. The column temperature was 35℃ and the UV detection wavelength was at 265 nm. The method was used for determination of the related substances which may exist in the L-orninthine monhydrochloride. Derivatization methods and conditions: The derivative solution which contained sample, 0.05 mol·L-1 borate buffer solution and 8 mg·mL-1 FMOC-Cl acetonitrile solution (v/v, 1∶3∶3) was reacted for 20 minutes at room temperature. Then, n-hexane was used twice to remove derivative reagents. After that, the derivative solution pH was adjusted to 3-4 by 4 mol·L-1 hydrochloric acid solution. Results: L-Ornithine monohy-drochloride and other 12 potentially related amino acids achieved good separations. Linear calibrations were established for all the tested amino acids in the range from 1.0μg·mL-1 to 20μg·mL-1 (r>0.9995). The re-sults of standard recovery and precision were good. Conclusion: The method has good specificity and ac-curacy which is suitable for the determination of related substances of L-ornithine monohydrochloride.
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