Insect venom is a highly complex mixture of bioactive compounds, containing proteins, peptides, and small molecules. Environmental factors can alter the venom composition and lead to intraspecific variation in its bioactivity properties. The investigation of discriminating compounds caused by variation impacts can be a key to manage sampling and explore the bioactive compounds. The present study reports the development of a peptidomic methodology based on UHPLC-ESI-QTOF-HRMS analysis followed by a nontargeted multivariate analysis to reveal the profile variance of Vespa velutina venom collected in different conditions. The reliability of the approach was enhanced by optimizing certain XCMS data processing parameters and determining the sample peak threshold to eliminate the interfering features. This approach demonstrated a good repeatability and a criterion coefficient of variation (CV) > 30% was set for deleting nonrepeatable features from the matrix. The methodology was then applied to investigate the impact of collection period variation. PCA and PLS-DA models were used and validated by cross-validation and permutation tests. A slight discrimination was found between winter and summer hornet venom in two successive years with 10 common discriminating compounds. Graphical abstract.

中文翻译：

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LC-MS多元非靶向方法的开发，用于对亚洲大黄蜂毒液（Vespa velutina nigrithorax）的肽谱进行差异分析：在研究收集期变异的影响中的应用。

昆虫毒液是生物活性化合物的高度复杂混合物，包含蛋白质，肽和小分子。环境因素会改变毒液成分，并导致其生物活性特性发生种内变化。研究由变异影响引起的区分化合物可能是管理采样和探索生物活性化合物的关键。本研究报告了基于UHPLC-ESI-QTOF-HRMS分析的肽组学方法的发展，然后进行了非目标多变量分析，以揭示在不同条件下收集的Vespa velutina毒液的轮廓差异。通过优化某些XCMS数据处理参数并确定样本峰值阈值以消除干扰功能，可以提高该方法的可靠性。这种方法证明了良好的可重复性，并且设置了标准变异系数（CV）> 30％，用于从矩阵中删除不可重复的特征。然后将该方法应用于调查收集期变化的影响。使用了PCA和PLS-DA模型，并通过交叉验证和置换测试对其进行了验证。在连续两年中，在冬季和夏季大黄蜂毒液之间发现了轻微的区别，其中包含10种常见的区别化合物。图形概要。在连续两年中，在冬季和夏季大黄蜂毒液之间发现了轻微的区别，其中包含10种常见的区别化合物。图形概要。在连续两年中，在冬季和夏季大黄蜂毒液之间发现了轻微的区别，其中包含10种常见的区别化合物。图形概要。