半夏提取物中尿苷含量超高效液相色谱检测法的建立及半夏不同提取工艺的比较

Development of ultra performance liquid chromatography for uridine content in Pinellia ternata extract and comparison of various extraction procedures

目的建立测定半夏提取物中尿苷含量的超高效液相色谱（ultra performance liquid chromatography，UPLC）法，并进行验证，同时对半夏不同提取工艺的尿苷含量进行比较。方法应用UPLC法测定尿苷对照品溶液、供试品溶液中尿苷的含量，并对方法的线性、精密度、稳定性、重复性、准确性进行验证。采用建立的方法检测传统水提醇沉工艺及隔膜压滤工艺半夏提取物中尿苷含量。结果半夏提取物中尿苷在0．4116—20．58μg／ml浓度范围内，与峰面积呈良好的线性关系（r=0．9999）；用建立的方法连续进样6次，检测对照品溶液中尿苷含量，RSD=0．32％；室温下放置8h，检测对照品中尿苷含量，RSD=0．23％；隔膜压滤工艺半夏提取液取6份进样，检测的尿苷平均含量为0．016％，RSD=0．65％；9份隔膜压滤工艺半夏提取液的平均加标回收率为99．25％，RSD=1．06％。隔膜压滤工艺的指纹图谱特征峰数目多于传统水提醇沉工艺，核苷类成分的含量也明显高于水提醇沉工艺；隔膜压滤工艺中尿苷含量为传统水提醇沉工艺的13倍。结论建立了检测半夏提取物中尿苷含量的UPLC法，该方法精密度、稳定性、重复性及准确性好，为科学有效控制半夏药材及饮片的质量提供了参考。

Objective To develop and verify a uhra performance liquid chromatography （UPLC） method for uridine con-tent in Pinelia ternate and compare the uridine contents in extracts by various procedures. Methods The uridine contents in reference and test samples were determined by UPLC, and the method was verified for linearity, precision, stability, reproducibility and accuracy. The uridine contents in Pinelia ternate extracted by traditional water extraction and alcohol precipitation procedure and by membrane filter press extraction. Results The uridine content in Pinelia ternate extract showed good linear relationship to peak area at a range of 0. 411 6 - 20. 58 txg/ml （r = 0. 999 9）. The RSD of determi-nation results of reference loaded for 6 times was 0. 32%, while that of reference after storage at room temperature for 8 h was 0. 23%. The mean uridine content in 6 test samples of Pinelia ternate by membrane filter press extraction was 0. 016%, with a RSD of 0. 65%. However, the average recovery of 9 test samples of Pinelia ternate by membrane filter press extraction was 99. 25%, with a RSD of 1.06%. The number of peak on fingerprint chromatogram of Pinelia ternate by membrane filter press extraction was larger than that by traditional water extraction and alcohol precipitation proce-dure, while the nucleoside content of the former was significantly higher than that of the latter. The uridine content of the former was 13 times of that of the latter. Conclusion The UPLC method for uridine content in Pinelia ternate was de-veloped, which showed high precision, stability, reproducibility and accuracy, which provided a reference for scientific and effective quality control of material and tablets of Pinelia ternate.