Co-occurrences of peptides and chemical components are usually observed in complicated matrices. Notably, those traditional Chinese medicine prescriptions (TCMPs) contain both plant and animal ingredients. It is still challenging to simultaneously monitor peptides and chemical components attributing to their different liquid chromatographic (LC) and mass spectrometric (MS) behaviors. Herein, efforts were made to pursue an eligible approach enabling simultaneous determination of peptides and chemical components in a TCMP namely Cervus and Cucumis polypeptide injection (CCPI, Songmeile®) that is prepared from the acid hydrolytic peptide-enriched extract of Sika deer (Cervus nippon Temminck) bone and the aqueous extract of muskmelon (Cucumis melo L.) seeds. Reversed phase liquid chromatography and hydrophilic interaction liquid chromatography were serially connected (RPLC-HILIC) to achieve comprehensive retention and separation. Sensitive detection was accomplished with selected reaction monitoring (SRM) mode, and multiply charged and singly charged ion transitions were defined for peptides and chemical components, respectively. Inter-batch variations of CCPI were evaluated in an authentic compound-independent manner. In particular, online energy-resolved MS was proposed to gain optimal parameters for five targeted peptides after that CCPI peptidome was profiled using nanoLC-LTQ Orbitrap Velos Pro MS. A so-called universal metabolome standard (UMS) sample was built for calibration curve construction and subsequently applied to acquire the quasi-contents of all 31 analytes, including five peptides and 26 chemical components, in ten batches of CCPI (CCPI1-CCPI10). The quantitative dataset revealed mild fluctuation for the quasi-content profiles of analytes-of-interest within different batches. More importantly, RPLC-HILIC-SRM is a promising method to fully address the demands of simultaneous measurement of peptides and chemical components in complicated matrices, and it might be a robust analytical tool for in-depth quality evaluation of CCPI as well as other TCMPs.

中文翻译：

---

使用反相液相色谱-亲水相互作用液相色谱-串联质谱法同时定量分析宫颈和黄瓜多肽注射液中的肽和化学成分（Songmeile®）。

通常在复杂的基质中会同时出现肽和化学成分。值得注意的是，那些传统中药处方（TCMPs）包含植物和动物成分。同时监测归因于其不同液相色谱（LC）和质谱（MS）行为的肽和化学成分仍然具有挑战性。在此，我们努力寻求一种合格的方法，该方法能够同时测定TCMP中的肽和化学成分，即由富含酸水解肽的梅花鹿（Cervus nippon）提取的鹿和黄瓜多肽注射液（CCPI，Songmeile®）。 Temminck）骨头和甜瓜（Cucumis melo L.）种子的水提物。反相液相色谱和亲水相互作用液相色谱串联（RPLC-HILIC）以实现全面的保留和分离。灵敏的检测是通过选择的反应监测（SRM）模式完成的，分别为肽和化学成分定义了多电荷和单电荷离子跃迁。CCPI的批间变异以可靠的独立于化合物的方式进行了评估。特别是，在使用nanoLC-LTQ Orbitrap Velos Pro MS对CCPI肽组进行了分析后，提出了在线能量分辨MS以获得五个目标肽的最佳参数。建立了所谓的通用代谢组标准品（UMS）样品，用于校准曲线的构建，随后用于获取所有31种分析物的准含量，十批CCPI（CCPI1-CCPI10）中包括5种肽和26种化学成分。定量数据集揭示了不同批次内目标分析物的准含量曲线的轻微波动。更重要的是，RPLC-HILIC-SRM是一种有前途的方法，可以完全满足同时测量复杂基质中肽和化学成分的需求，并且它可能是用于CCPI和其他TCMP的深入质量评估的强大分析工具。 。