反相高效液相色谱法测定壳聚糖纳米粒中丙酮酸乙酯的含量

摘要

背景：有关丙酮酸乙酯含量检测方法的研究较少，采用反相高效液相色谱法检测丙酮酸乙酯含量的文献更少。目的：建立测定壳聚糖纳米粒中丙酮酸乙酯含量的反相高效液相色谱法。　　方法：采用Agilent1200系列高效液相色谱仪检测丙酮酸乙酯-壳聚糖纳米粒中丙酮酸乙酯的含量。色谱柱：ZORBAX Eclipse XDB-C18(4.6 mm×150 mm，5μm)，柱温25℃，流动相为乙腈-水(体积比为40∶60)，流速1 mL/min，检测波长210 nm，进样量20μL。　　结果与结论：丙酮酸乙酯峰与辅料及溶剂峰分离良好，丙酮酸乙酯质量浓度在1-100 mg/L内于峰面积线性关系良好(r =0.9996)，低、中、高浓度丙酮酸乙酯对照品溶液日内、日间精密度的相对标准偏差均小于3%，重复性实验的相对标准偏差为1.25%，稳定性实验的相对标准偏差为1.3%。3批样品的加样回收率分别为(91.5±1.0)%，(93.5±0.2)%，(94.4±0.4)%；包封率分别为(87.20±0.22)%，(90.50±0.15)%，(91.10±0.17)%。不同批次样品丙酮酸乙酯含量检测结果的相对标准偏差分别为0.9%，0.5%，0.3%。说明反相高效液相色谱法灵敏度高、线性范围宽、专属性强、精密度高、加样回收率好、结果精确可靠，可用于壳聚糖纳米粒中丙酮酸乙酯含量的测定。%BACKGROUND:Research on ethyl pyruvate detection methods is reported rarely, and moreover, literature about reversed-phase high-performance liquid chromatography (RP-HPLC) for detection of ethyl pyruvate is less. OBJECTIVE:To establish an RP-HPLC method for determination of ethyl pyruvate in ethyl pyruvate-chitosan nanoparticles. METHODS: The chromatographic analysis was performed on a ZORBAX Eclipse XDB-C18 column (4.6 mm× 150 mm, 5μm) at 25℃, with the mixture of acetonitrile and water (40:60, V/V) as the mobile phase at the flow rate of 1 mL/min. The determination wavelength wasset at 210 nm and the injection volume was 20 μL. RESULTS AND CONCLUSION: The peak of ethyl pyruvate and the peaks of auxiliary materials and solvent were separated wel. The linear rang of ethyl pyruvate was 1-100 mg/L (r=0.999 6). The relative standard deviation of both the intra-and inter-day precision was less than 3% for low-, moderate-, and high-concentration ethyl pyruvate. The relative standard deviation of reproducibility test and stability test was 1.25% and 1.3%, respectively. Sample average recovery rates were (91.5±1.0)%, (3.5±0.2)%, (94.4±0.4)%, respectively. Encapsulation efficiency of samples were (87.2±0.22)%, (90.5±0.15)%, (91.1±0.17)%, respectively. The relative standard deviation of different sample content were 0.9%, 0.5%, 0.3%, respectively. The RP-HPLC method for determination of ethyl pyruvate is sensitive, accurate and highly specific with wide linear range and high sample average recovery.