目的 建立阿胶UHPLC特征图谱及游离氨基酸含量测定方法，并结合指纹图谱相似度评价、化学计量学对不同批次的阿胶进行质量评价。方法 采用Waters BEH C₁₈（100 mm×2.1 mm，1.7 µm）柱，以0.1%甲酸水（A）-乙腈（B）为流动相，梯度洗脱，体积流量为0.3 mL/min，检测波长254 nm，柱温40 ℃，进样量2 μL。对14批阿胶进行特征图谱建立，确定共有峰并进行相似度评价；基于特征图谱共有峰面积测定结果，结合化学计量学手段对14批阿胶进行质量评价；并对甘氨酸（Gly）、L-脯氨酸（Pro）、L-丙氨酸（Ala）、L-甲硫氨酸（Met）、L-缬氨酸（Val）、L-亮氨酸（Leu）、L-苯丙氨酸（Phe）进行含量测定。结果 指纹图谱相似度评价结果表明，14批阿胶样品的相似度均≥0.927，说明各批次之间相似度良好，质量较稳定，共标定了9个共有峰，并指认了Gly、Pro、Ala、Met、Val、Leu、Phe 7个共有峰；化学计量学结果将14批阿胶划分为3类并筛选出导致不同批次间差异的差异性标志物为Met、Leu和Gly；Gly、Pro、Ala、Met、Val、Leu、Phe 7个成分的质量分数分别在319.8～1 227.27 µg/g、100.85～352.44 µg/g、113.92～531.06 µg/g、50.57～182.8 µg/g、47.5～116.53 µg/g、48.64～182.37 µg/g、45.65～124.69 µg/g。结论 建立的方法快捷，准确可靠，可为阿胶质量控制进一步提升提供实验参考。

Objective A method was developed to establish a UHPLC characteristic spectrum and determine the free amino acid content of Ejiao (Colla Corii Asini). The quality of different batches of Colla Corii Asini was assessed by combining fingerprint spectrum similarity evaluation and chemometrics.Methods UHPLC was launched on a Waters BEH C₁₈ (100 mm×2.1 mm, 1.7 µm) column by gradient elution with a mobile phase of 0.1% formic acid water (A) and acetonitrile (B) at a flow rate of 0.3 mL/min, detection wavelength of 254 nm, column temperature of 40 ℃, and an injection volume of 2 μL. The characteristic spectrum of 14 batches of Colla Corii Asini was established, common peak was identified and a similarity evaluation was carried out. Based on the determination results of the common peak area in the characteristic spectra, the quality of 14 batches of Colla Corii Asini was evaluated by combining chemometrics methods. The levels of glycine (Gly), L-proline (Pro), L-alanine (Ala), L-methionine (Met), L-valine (Val), L-leucine (Leu), and L-phenylalanine (Phe) were quantified.Results The results of the fingerprint similarity evaluation showed that the similarity of 14 batches of Colla Corii Asini samples was ≥ 0.927, indicating a good similarity and stable quality. A total of nine common peaks were calibrated and seven common peaks were identified, namely Gly, Pro, Ala, Met, Val, Leu and Phe. The chemometrics analysis classified 14 batches of Asini Corii Colla into three categories, identifying the differential markers that caused differences between different batches as Met, Leu and Gly. The mass concentrations of the seven components, Gly, Pro, Ala, Met, Val, Leu and Phe, range from 319.8—1227.27 µg/g, 100.85—352.44 µg/g, 113.92—531.06 µg/g, 50.57—182.8 µg/g, 47.5—116.53 µg/g, 48.64—182.37 µg/g, 45.65—124.69 µg/g.Conclusion The method established in this study is fast, accurate and reliable, which can provide an experimental reference for further research on improving the quality standards of Colla Corii Asini.

R286.2

贵州省科技计划项目（黔科合重大专项字[2024]015，黔科合平台人才-GCC[2023]036，黔科合平台人才-CXTD[2023]019，黔科合平台人才-GCC[2022]031-1）；贵阳市科技计划项目（筑科合同[2024]2-35号）