Proteomic Analysis and the Antioxidant Activity Evaluation of Asini Corii Colla Powder Based on Liquid Chromatography-Tandem Mass Spectrometry Technology

doi:10.19894/j.issn.1000-0518.250344

Chinese Journal of Applied Chemistry ›› 2026, Vol. 43 ›› Issue (3): 396-410.DOI: 10.19894/j.issn.1000-0518.250344

• Full Papers • Previous Articles Next Articles

Proteomic Analysis and the Antioxidant Activity Evaluation of Asini Corii Colla Powder Based on Liquid Chromatography-Tandem Mass Spectrometry Technology

Xiao XIE¹, Cong LI¹, Jian-Ling ZHANG¹, Zhi-Jie CHEN¹, Chun-Xiang TAO², Mei-Mei FU³(), Hui-Lin YOU⁴, Jin-Shan GUO²^,³^,⁴()

^1.Liaocheng Key Laboratory of Functional Food Research and Development，Liaocheng Engineering Research Center for Nutrition and Health Consumer Products，Dong'e Ejiao Health Industry Technology Co. ，Ltd. ，Liaocheng 252201，China
^2.CAS Key Laboratory of High-Performance Synthetic Rubber and Its Composite Materials，Changchun Institute of Applied Chemistry，Chinese Academy of Sciences，Changchun 130022，China
^3.Department of Histology and Embryology，NMPA Key Laboratory for Safety Evaluation of Cosmetics，GDMPA Key Laboratory of Key Technologies for Cosmetics Safety and Efficacy，School of Basic Medical Sciences，Southern Medical University，Guangzhou 510515，China
^4.Regenerative Medicine and Tissue Repair Research Center，Huangpu Institute of Materials，Guangzhou 511363，China

Received:2025-09-03 Accepted:2025-11-28 Published:2026-03-01 Online:2026-03-26
Contact: Mei-Mei FU,Jin-Shan GUO
About author:jsguo4127@163.com
fmm05037726@163.com；
Supported by:
the Industry-sponsored Project with Dong'e Ejiao Health Products Co., Ltd(DEEJ-YB-2024-10-190)

Abstract

Abstract:

Asini corii colla contains multiple bioactive proteins， endowing it with multifaceted health benefits. This study employed liquid chromatography-tandem mass spectrometry （LC-MS/MS） to systematically identify and analyze the protein composition of Asini corii colla powder， identifying various functional proteins including multiple intermediate filament proteins， collagens， immunomodulatory proteins， and antioxidant-related proteins. Further evaluation of Asini corii colla powder's cytotoxicity， proliferation， and migration effects was conducted using human dermal fibroblasts （HSF）. Results indicated no significant cytotoxicity within the tested concentration range （0.156~20 mg/mL）， while promoting HSF proliferation and migration， suggesting preliminary potential for Asini corii colla powder in skin repair applications. To investigate the antioxidant stress activity of Asini corii colla， this study further employed a hydrogen peroxide （H?O?） induced cellular oxidative stress model to evaluate the effects of Asini corii colla powder on intracellular reactive oxygen species （ROS） scavenging capacity and antioxidant enzyme activity. Experiments demonstrated that pre-treatment with Asini corii colla powder significantly suppressed H?O?-induced ROS levels while enhancing intracellular superoxide dismutase （SOD）， catalase （CAT）， and glutathione peroxidase （GSH-Px） activities. This confirms its antioxidant effects are associated with strengthening the endogenous antioxidant defense system. Proteomics analysis further revealed that Asini corii colla powder may regulate oxidative stress by modulating the expression of proteins associated with the heat shock protein B family， biliverdin reductase B， hemoglobin subunit α， and nuclear factor κB （NF-κB） signaling pathways， thereby mitigating cellular oxidative stress damage to a certain extent. In summary， this study elucidates the protein composition of Asini corii colla and validates its role and mechanisms in combating oxidative stress， laying a scientific foundation for expanding its applications in skin repair and anti-aging.

Key words: Asini corii colla, Proteomics, Reactive oxygen species scavenge, Oxidative stress

CLC Number:

O636

Xiao XIE, Cong LI, Jian-Ling ZHANG, Zhi-Jie CHEN, Chun-Xiang TAO, Mei-Mei FU, Hui-Lin YOU, Jin-Shan GUO. Proteomic Analysis and the Antioxidant Activity Evaluation of Asini Corii Colla Powder Based on Liquid Chromatography-Tandem Mass Spectrometry Technology[J]. Chinese Journal of Applied Chemistry, 2026, 43(3): 396-410.

Figures/Tables 11

References 28

[1]	杨帅，鲁婷婷，周祖英，等. 阿胶化学成分和药理作用及质量控制研究进展［J］. 中国新药杂志， 2023， 32（8）： 806-816.
	YANG S， LU T T， ZHOU Z Y， et al. Research progress on chemical components， pharmacological effects and quality control of Asini corii colla［J］. Chin New Drug J， 2023， 32（8）： 806-816.
[2]	曲媛鑫，付英杰. 阿胶化学成分、质量控制及药理作用研究进展［J］. 特产研究， 2023， 45（3）： 136-143.
	QU Y X， FU Y J. Research progress on chemical components， quality control and pharmacological effects of Asini corii colla［J］. Spec Wild Eco Anim Plant Res， 2023， 45（3）： 136-143.
[3]	SANTOS A D， AMARAL JUNIOR A T D， FRITSCHE-NETO R， et al. Proteome analysis of phytomonas serpens， a phytoparasite of medical interest［J］. PLoS One， 2018， 13（10）： e0204818.
[4]	杨松柏，尹健，侯旭晖. 中药复杂体系的多组学分析技术研究进展［J］. 特产研究， 2024， 46（6）： 178-185.
	YANG S B， YIN J， HOU X H. Research progress on multi-omics analysis technologies for traditional chinese medicine complex systems［J］. Spec Wild Eco Anim Plant Res， 2024， 46（6）： 178-185.
[5]	李青，许伟沂，彭丽诗，等. 人参和西洋参提取物中皂苷组分比例分析及其在参类产品鉴别中的应用研［J］. 食品科技， 2024， 49（5）： 225-233.
	LI Q， XU W Y， PENG L S， et al. Analysis of saponin component proportions in extracts of panax ginseng and american ginseng and their application in identification of ginseng products［J］. Food Sci Technol， 2024， 49（5）： 225-233.
[6]	郝宏伟，陈意琪，何姣，等. 超高效液相色谱-电喷雾串联四级杆质谱鉴定灵芝中脂质成分［J］. 菌物学报， 2023， 42（5）： 1175-1184.
	HAO H W， CHEN Y Q， HE J， et al. Identification of lipid components in Ganoderma lucidum by ultra-performance liquid chromatography-electrospray ionization tandem quadrupole mass spectrometry［J］. Mycosystema， 2023， 42（5）： 1175-1184.
[7]	孙思阳，吕伟超，陈刚，等. 冬虫夏草的多成分含量综合评价分析［J］. 中国药剂学杂志， 2025， 23（2）： 100-108.
	SUN S Y， LV W C， CHEN G， et al. Comprehensive evaluation and analysis of multiple component contents in cordyceps sinensis［J］. Chin Pharm J， 2025， 23（2）： 100-108.
[8]	GUO J， HUANG X， DOU L， et al. Aging and aging-related diseases：from molecular mechanisms to interventions and treatments［J］. Signal Transduction Targeted Ther， 2022， 7（1）： 391.
[9]	PAPACCIO F， ARINO A D， CAPUTO S， et al. Focus on the contribution of oxidative stress in skin aging［J］. Antioxidants， 2022， 11（6）： 1121.
[10]	BAI R， GUO J， YE X Y， et al. Oxidative stress： the core pathogenesis and mechanism of Alzheimer′s disease［J］， Ageing Res Rev， 2022， 77： 101619.
[11]	FORSTERMANN U， XIA N， LI H. Roles of vascular oxidative stress and nitric oxide in the pathogenesis of atherosclerosis［J］. Circ Res， 2017， 120（4）： 713-735.
[12]	NTASI G， SBRIGLIA S， PITOCCHI R A O， et al. Proteomic characterization of collagen-based animal glues for restoration［J］. J Proteome Res， 2022， 21（9）： 2173-2184.
[13]	廖峰，樊雨梅，帖航，等. 阿胶蛋白质组学研究［J］. 食品工业科技， 2021， 42（10）： 122-129.
	LIAO F， FAN Y M， TIE H， et al. Proteomic study of Asini corii colla［J］. Sci Technol Food Ind， 2021， 42（10）： 122-129.
[14]	林伟欣，李树奇，杨莹莹，等. 基于4D-label-free技术的鹿皮胶与阿胶蛋白组分的对比研究［J］. 食品工业， 2024， 45（8）： 250-254.
	LIN W X， LI S Q， YANG Y Y， et al. Comparative study on protein components of deer skin glue and Asini corii colla based on 4D-label-free technology［J］. Food Ind， 2024， 45（8）： 250-254.
[15]	李聪，季萌萌，孟苓凤，等. 阿胶和阿胶糕不同极性溶剂提取物抗皮肤衰老功效探究［J］. 食品与发酵工业， 2025， 51（17）： 82-92.
	LI C， JI M M， MENG L F， et al. Investigation on the anti-aging effects of different polarity solvent extracts of Asini corii colla and asini corii colla cake on skin［J］. Food Ferment Ind， 2025， 51（17）： 82-92.
[16]	绰尔鹏，孟苓凤，李聪，等. 阿胶糕提取物对小鼠黑色素瘤细胞代谢的影响［J］. 食品科技， 2025， 50（4）： 222-230.
	CHUO E P， MENG L F， LI C， et al. Effect of Asini corii colla cake extract on metabolism of mouse melanoma cells［J］. Food Sci Technol， 2025， 50（4）： 222-230.
[17]	段小波，杜芬，王延涛，等. 阿胶和阿胶肽对血管内皮细胞氧化损伤的作用研究［J］. 安徽农业科学， 2025， 53（8）： 162-167， 180.
	DUAN X B， DU F， WANG Y T， et al. Study on the effect of Asini corii colla and Asini corii colla peptide on oxidative damage of vascular endothelial cells［J］. Anhui Agric Sci Bull， 2025， 53（8）： 162-167， 180.
[18]	FU M M， ZHAO Y T， WANG Y， et al. On-demand removable self-healing and pH-responsive europium-releasing adhesive dressing enables inflammatory microenvironment modulation and angiogenesis for diabetic wound healing［J］. Small， 2023， 19（3）： e2205489.
[19]	WU M， ZHAO Y T， TAO M H， et al. Malate-based biodegradable scaffolds activate cellular energetic metabolism for accelerated wound healing［J］. ACS Appl Mater Interfaces， 2023， 15（44）： 50836-50853.
[20]	WU K K， FU M M， ZHAO Y T， et al. Anti-oxidant anti-inflammatory and antibacterial tannin-crosslinked citrate-based mussel-inspired bioadhesives facilitate scarless wound healing［J］. Bioact Mater， 2023， 20： 93-110.
[21]	WANG Y， ZHAO Y T， MA S Y， et al. Injective programmable proanthocyanidin-coordinated zinc-based composite hydrogel for infected bone repair［J］. Adv Healthc Mater， 2024， 13（6）： e2302690.
[22]	GU T， ZHOU J， LIU Y， et al. Faucicola mancuniensis isolated from human respiratory tract and its genomic characteristicstics［J］. BMC Microbiol， 2025， 25（1）： 587.
[23]	ZOU S Q， ZHU X F， ZHANG L R， et al. Biomineralization-inspired synthesis of cerium-doped carbonaceous nanoparticles for highly hydroxyl radical scavenging activity［J］. Nanoscale Res Lett， 2018， 13（1）： 76.
[24]	SONG X L， ZHANG J J， LI J， et al. Acetylated polysaccharides from pleurotus geesteranus alleviate lung injury via regulating NF-κB signal pathway［J］. Int J Mol Sci， 2020， 21（8）： 2810.
[25]	ZHANG Q， YANG W B， LIU J C， et al. Identification of six flavonoids as novel cellular antioxidants and their structure-activity relationship［J］. Oxid Med Cell Longev， 2020， 2020： 4150897.
[26]	CHENG C S， GU Q H， ZHANG J K， et al. Phenolic constituents， antioxidant and cytoprotective activities， enzyme inhibition abilities of five fractions from vaccinium dunalianum wight［J］. Molecules， 2022， 27（11）： 3432.
[27]	ZHAO X， CUI Y J， BAI S S， et al. Antioxidant activity of novel casein-derived peptides with microbial proteases as characterized via Keap1-Nrf2 pathway in HepG2 cells［J］. J Microbiol Biotechnol， 2021， 31（8）： 1163-1174.
[28]	SHEN Y， SHEN Z， LI P， et al. Protective activity of Malus doumeri leaf extract on H₂O₂-induced oxidative injury in H9C2 rat cardiomyocytes［J］. Front Cardiovasc Med， 2022， 9： 1005306.

Pathway ID	Proteins	Number of peptide segments	Peptide matching map	Peptide relative molecular mass	Peptide sequence
P01959	Hemoglobin subunit alpha	1	1	15 192.9	FLSTVSTVLTSK
P35747	Albumin	1	1	68 536.2	TVFDQFTPLVEEPK
A0A3Q2HPI8	Heat shock protein B family member 6（HSPB6）	2	2	16 996.8	HEERPDEHGYIAR；VVGEHVEVHAR
A0A3Q2IC86	EF-hand domain-containing protein	2	2	188 977.1	EFGDVLR； QYLEEEEQLQR
A0A8C4KTB8	Protein S100	1	1	11 305.5	TEAEASVIGIIELFHK
A0A8C4KUI5	Protein S100	2	2	11 734.7	ALDVMVSTFHK；ELPSFLGK
A0A8C4KW94	Desmoplakin	1	1	98 434.9	VMGNQVFPEVTR
A0A8C4L337	Keratin 39	2	2	55 591.6	ADLEAQLQSLK； YEAEVSLR
A0A8C4L453	Immunoglobulin-like domain-containing protein	1	1	42 567.6	EPQVYVLAPHPDELSK
A0A8C4L8M0	Tropomyosin 1	1	1	40 872.8	SIDDLEDELYAQK
A0A8C4LCA2	Collagen type Ⅵ alpha 2 chain	2	2	107 325.2	DYDSLAQPSFFDR； EKDYDSLAQPSFFDR
A0A8C4LF88	BPI fold containing family A member 2	1	1	26 938.4	ASLELQTDVR
A0A8C4LHC7	Keratin， type Ⅰ cytoskeletal 14	1	1	52 113.7	VVSTHEQVLR
A0A8C4LKF1	Keratin， type Ⅰ cytoskeletal 17	1	1	48 016.9	TIVEEVQDGR
A0A8C4LSE8	IF rod domain-containing protein	3	3	53 274.3	RYEEEVSLR； WQLYQNR； YEEEVSLR
A0A8C4LSH3	Keratin 85	3	3	60 475.0	KSDLEANVEALVEESSFLK； LEAAVAEAEQQGEAALTDAR； SDLEANVEALVEESSFLK
A0A8C4LSI1	Keratin 84	1	1	62 358.4	MAAVGPHPIR
A0A8C4M581	Myosin-2	1	1	222 648.8	TLALLFSGAQTADAEAGGGVK
A0A8C4MCM7	Desmocollin 4	1	1	113 727.6	IISQEPAGAPMFILNR
A0A8C4PFM4	Globin domain-containing protein	1	1	16 182.3	FFDSFGDLSNPAAVMGNPK
A0A8C4PJ20	Keratin 15	1	1	49 402.4	LAGIGTGEVSLGGGSGGK
A0A8C4PTT8	Binding immunoglobulin Protein	1	1	72 164.4	FEELNMDLFR
A0A9L0J7I9	Collagen type Ⅵ alpha 1 chain	1	1	108 945.3	VPSYQELLDGVFYQSVSR
A0A9L0RZQ4	Collagen type Ⅰ alpha 2 chain	1	1	116 870.2	MGLMGPR
A0A9L0TQW9	IF rod domain-containing protein	1	1	59 937.5	LEAAVSQAEQQGEAALSDAK
F6X6A6	Peptidyl-prolyl cis-trans isomerase	1	1	17 855.7	VNPTVFFDIAVDGEPLGR
A0A9L0JCK5	IF rod domain-containing protein	1	1	53 832.2	LEVAVSQAEQQGEAALSDAK
A0A8C4L2C5	Small ribosomal subunit protein eS28	1	1	7 818.1	EGDVLTLLESER
F7CYP1	Serine protease inhibitor domain-containing protein	1	1	46 822.1	GTEAAGATIVEAIR
A0A3Q2I2H1	Immunoglobulin-like domain-containing protein	1	1	10 624.0	PGGPPVTVIYK
A0A8C4MBM1	Vasodilator-stimulated phosphoprotein	1	1	40 502.3	EEIIEAFVQELR
Q6X9Z5	Large ribosomal subunit protein P2	1	1	11 696.8	NIEDVIAQGIGK
A0A3Q2KYZ9	Keratin-associated protein 7-1	1	1	9 125.1	PWGSGSGFGYSTY
A0A9L0JDB8	Biliverdin reductase B	1	1	21 957.3	IALFGATGR
A0A8C4L4K0	SH3 domain-containing protein	1	1	321 654.4	LLAPLEIAK

Pathway ID	Proteins	Number of peptide segments	Peptide matching map	Peptide relative molecular mass	Peptide sequence
P01959	Hemoglobin subunit alpha	1	1	15 192.9	FLSTVSTVLTSK
P35747	Albumin	1	1	68 536.2	TVFDQFTPLVEEPK
A0A3Q2HPI8	Heat shock protein B family member 6（HSPB6）	2	2	16 996.8	HEERPDEHGYIAR；VVGEHVEVHAR
A0A3Q2IC86	EF-hand domain-containing protein	2	2	188 977.1	EFGDVLR； QYLEEEEQLQR
A0A8C4KTB8	Protein S100	1	1	11 305.5	TEAEASVIGIIELFHK
A0A8C4KUI5	Protein S100	2	2	11 734.7	ALDVMVSTFHK；ELPSFLGK
A0A8C4KW94	Desmoplakin	1	1	98 434.9	VMGNQVFPEVTR
A0A8C4L337	Keratin 39	2	2	55 591.6	ADLEAQLQSLK； YEAEVSLR
A0A8C4L453	Immunoglobulin-like domain-containing protein	1	1	42 567.6	EPQVYVLAPHPDELSK
A0A8C4L8M0	Tropomyosin 1	1	1	40 872.8	SIDDLEDELYAQK
A0A8C4LCA2	Collagen type Ⅵ alpha 2 chain	2	2	107 325.2	DYDSLAQPSFFDR； EKDYDSLAQPSFFDR
A0A8C4LF88	BPI fold containing family A member 2	1	1	26 938.4	ASLELQTDVR
A0A8C4LHC7	Keratin， type Ⅰ cytoskeletal 14	1	1	52 113.7	VVSTHEQVLR
A0A8C4LKF1	Keratin， type Ⅰ cytoskeletal 17	1	1	48 016.9	TIVEEVQDGR
A0A8C4LSE8	IF rod domain-containing protein	3	3	53 274.3	RYEEEVSLR； WQLYQNR； YEEEVSLR
A0A8C4LSH3	Keratin 85	3	3	60 475.0	KSDLEANVEALVEESSFLK； LEAAVAEAEQQGEAALTDAR； SDLEANVEALVEESSFLK
A0A8C4LSI1	Keratin 84	1	1	62 358.4	MAAVGPHPIR
A0A8C4M581	Myosin-2	1	1	222 648.8	TLALLFSGAQTADAEAGGGVK
A0A8C4MCM7	Desmocollin 4	1	1	113 727.6	IISQEPAGAPMFILNR
A0A8C4PFM4	Globin domain-containing protein	1	1	16 182.3	FFDSFGDLSNPAAVMGNPK
A0A8C4PJ20	Keratin 15	1	1	49 402.4	LAGIGTGEVSLGGGSGGK
A0A8C4PTT8	Binding immunoglobulin Protein	1	1	72 164.4	FEELNMDLFR
A0A9L0J7I9	Collagen type Ⅵ alpha 1 chain	1	1	108 945.3	VPSYQELLDGVFYQSVSR
A0A9L0RZQ4	Collagen type Ⅰ alpha 2 chain	1	1	116 870.2	MGLMGPR
A0A9L0TQW9	IF rod domain-containing protein	1	1	59 937.5	LEAAVSQAEQQGEAALSDAK
F6X6A6	Peptidyl-prolyl cis-trans isomerase	1	1	17 855.7	VNPTVFFDIAVDGEPLGR
A0A9L0JCK5	IF rod domain-containing protein	1	1	53 832.2	LEVAVSQAEQQGEAALSDAK
A0A8C4L2C5	Small ribosomal subunit protein eS28	1	1	7 818.1	EGDVLTLLESER
F7CYP1	Serine protease inhibitor domain-containing protein	1	1	46 822.1	GTEAAGATIVEAIR
A0A3Q2I2H1	Immunoglobulin-like domain-containing protein	1	1	10 624.0	PGGPPVTVIYK
A0A8C4MBM1	Vasodilator-stimulated phosphoprotein	1	1	40 502.3	EEIIEAFVQELR
Q6X9Z5	Large ribosomal subunit protein P2	1	1	11 696.8	NIEDVIAQGIGK
A0A3Q2KYZ9	Keratin-associated protein 7-1	1	1	9 125.1	PWGSGSGFGYSTY
A0A9L0JDB8	Biliverdin reductase B	1	1	21 957.3	IALFGATGR
A0A8C4L4K0	SH3 domain-containing protein	1	1	321 654.4	LLAPLEIAK

Proteomic Analysis and the Antioxidant Activity Evaluation of Asini Corii Colla Powder Based on Liquid Chromatography-Tandem Mass Spectrometry Technology

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 11

References 28

Related Articles 3

Recommended Articles

Metrics

Comments

[1]	Xin XU, Ye LI, Min WANG, Zhang-Cheng FU, Guo-Min QI, Chun-Hua LU. Self‑Assembled MicroRNA‑21 Responsive Zn²⁺/DNA Nanoparticles for Tumor Fluorescence Imaging and Oxidative Stress Therapy [J]. Chinese Journal of Applied Chemistry, 2024, 41(1): 164-174.
[2]	GAO Ming, ZHAO Kaidong, LIU Xiangyong, JIN Yuanzhe. Preparation of Molybdenum Disulfide Nanoparticles and the Cytoprotection on Cardiac Myocytes [J]. Chinese Journal of Applied Chemistry, 2020, 37(9): 1010-1021.
[3]	XU Yulin,LIU Chunrong. Research Progress on Fluorescence Detection of Methionine Sulfoxide/Methionine Sulfoxide Reductases [J]. Chinese Journal of Applied Chemistry, 2018, 35(1): 21-27.

Time/min	A（0.2% acetonitrile-0.1% formic acid）/%	B（98% acetonitrile-0.1% formic acid）/%
0	95	5
2	95	5
43	75	25
47	65	35
49	20	80
51	20	80
60	95	5

Time/min	A（0.2% acetonitrile-0.1% formic acid）/%	B（98% acetonitrile-0.1% formic acid）/%
0	95	5
2	95	5
43	75	25
47	65	35
49	20	80
51	20	80
60	95	5