液相色谱-质谱联用鉴定抗菌肽Cbf-14凝胶的有关物质

霍怡彤; 徐柯卉; 陆宇婷; 马菱蔓; 周长林; 杭太俊; 宋敏

doi:10.11665/j.issn.1000-5048.20220509

液相色谱-质谱联用鉴定抗菌肽Cbf-14凝胶的有关物质

1.
中国药科大学药学院，南京 211198
2.
中国药科大学生命科学与技术学院，南京 211198

计量
- 文章访问数: 191
- HTML全文浏览量: 11
- PDF下载量: 371
出版历程
- 收稿日期: 2022-05-05
- 修回日期: 2022-09-13
- 刊出日期: 2022-10-24

Identification of the related substances of antimicrobial peptide Cbf-14 gel by LC-MS

1.
School of Pharmacy, China Pharmaceutical University, Nanjing 211198
2.
School of Life Science & Technology, China Pharmaceutical University, Nanjing 211198, China

摘要

摘要: 多肽Cbf-14是由14个氨基酸组成的新型抗菌肽。采用优化的反相高效液相色谱-电喷雾正离子化-四极杆飞行时间串联质谱（LC-ESI-QTOF/MS）检测方法，Agilent ZORBAX SB-Phenyl（150 mm × 4.6 mm，3.5 μm）色谱柱，以乙腈-20 mmol/L甲酸铵水溶液（甲酸调节pH为3.0）为流动相进行梯度洗脱，对Cbf-14凝胶中的有关物质进行了分离鉴定。在所建立的分析条件下，Cbf-14及其有关物质分离良好，检测并鉴定出Cbf-14凝胶及其强制降解试验样品中24个主要有关物质，其中5个为制剂生产工艺杂质，19个为降解产物，分析了有关物质的来源和多肽制剂的稳定性，为其生产工艺优化、贮存条件制定和质量控制提供了参考依据。
- 抗菌肽Cbf-14凝胶 /
- 有关物质 /
- 结构鉴定 /
- 液相色谱-质谱联用
Abstract: Cbf-14 is a novel antimicrobial peptide composed of 14 amino acids.An optimized reversed phase high-performance liquid chromatographic method with electrospray-ionization quadrupole time-of-flight mass spectrometer (LC-ESI-QTOF/MS) method was developed for separation, identification and characterization of structurally related peptide impurities in Cbf-14 gel.Chromatographic separation was carried out on an Agilent ZORBAX SB-Phenyl column (150 mm × 4.6 mm, 3.5 μm), with acetonitrile-20 mmol/L ammonium formate buffer (adjusted to pH 3.0 with formic acid) as eluent using gradient elution.Under the established conditions, Cbf-14 and its structurally related peptide impurities were well separated; and a total of 24 impurities were detected and identified, of which 5 were impurities in the preparation manufacturing process and 19 were stressed products.Based on high resolution mass spectrometry analysis, the origins and formation mechanisms of these impurities were located.The obtained results are useful for the establishment of the manufacturing process, storage condition and quality control of Cbf-14 gel.
- antimicrobial peptide Cbf-14 gel /
- related substances /
- structural identification /
- LC-MS

HTML全文

参考文献(13)

[1]	. Chin Stra Emer Indus（中国战略新兴产业），2017，15（29）：68-70.
[2]	Li GN，Xia XJ，Long YH，et al. Research progresses and applications of antimicrobial peptides［J］. Chin J Animal Nutr（动物营养学报），2014，26（1）：17-25.
[3]	Zhou HM，Dou J，Wang J，et al. The antibacterial activity of BF-30 in vitro and in infected burned rats is through interference with cytoplasmic membrane integrity［J］. Peptides，2011，32（6）：1131-1138.
[4]	Tian YW，Wang H，Li B，et al. The cathelicidin-BF Lys16 mutant Cbf-K16 selectively inhibits non-small cell lung cancer proliferation in vitro［J］. Oncol Rep，2013，30（5）：2502-2510.
[5]	Ma LM，Wang YR，Wang MX，et al. Effective antimicrobial activity of Cbf-14，derived from a cathelin-like domain，against penicillin-resistant bacteria［J］. Biomaterials，2016，87：32-45.
[6]	Yu CZ，Wei SS，Han XR，et al. Effective inhibition of Cbf-14 against Cryptococcus neoformans infection in mice and its related anti-inflammatory activity［J］. Fungal Genet Biol，2018，110：38-47.
[7]	Wang P. Analysis of key problems on synthesis of synthetic peptide drugs［J］. Chin J New Drugs（中国新药杂志），2010，19（2）：102-105.
[8]	Hu YX，Jiang Y，Han TJ. Effects of manufacturing process and process control on related substances of synthetic peptide drugs［J］. Chin J New Drugs（中国新药杂志），2017，26（18）：2143-2148.
[9]	Hu YX，Jiang Y，Han TJ，et al. Quality control and related substances of synthetic peptide drugs［J］. Chin J New Drugs（中国新药杂志），2018，27（5）：502-508.
[10]	D''Hondt M，Bracke N，Taevernier L，et al. Related impurities in peptide medicines［J］. J Pharm Biomed Anal，2014，101：2-30.
[11]	Tian WJ，Ren X，Liao HM，et al. Research progress of the quality control of peptide drugs［J］. Chin J Pharm Anal（药物分析杂志），2013，33（7）：1115-1120.
[12]	Requena JR，Chao CC，Levine RL，et al. Glutamic and aminoadipic semialdehydes are the main carbonyl products of metal-catalyzed oxidation of proteins［J］. Proc Natl Acad Sci U S A，2001，98（1）：69-74.
[13]	Stadtman ER. Metal ion-catalyzed oxidation of proteins：biochemical mechanism and biological consequences［J］. Free Radic Biol Med，1990，9（4）：315-325.

施引文献(9)

期刊类型引用(4)

1.	宋永真，李雅静，张振凌，王胜超. 曼地亚红豆杉酒炙工艺及质量标准研究. 中医学报. 2024(05): 1068-1075 . 百度学术
2.	剡槿熙，臧明伍，刘贺，徐晨晨，史宇璇，白京，赵燕，郝静怡，张亚茹. 二萜类化合物在肉类食品保鲜防腐中的应用研究进展. 食品科学. 2024(13): 282-291 . 百度学术
3.	王楠楠，蔡婷婷，童晔玲，刘霞，朱婉萍，江石平，赵桂芝. 基于NF-κB信号通路的HTRF法筛选红豆杉中抗气道炎症有效活性成分研究. 浙江中医杂志. 2023(08): 553-556 . 百度学术
4.	傅贵江. 杉木人工林抚育间伐对套种南方红豆杉的影响. 武夷科学. 2022(01): 39-43 . 百度学术