Accuracy and Precision Comparison for Molecular Weight Distribution Assay of Fish Collagen Peptides: a Methodology Study Between Two Gel Permeation Chromatography Columns
Two gel permeation chromatography columns, Superdex Peptide HR 10/30 column and TSK G2000 SWXL column, were compared for their accuracy and precision for determining the molecular weight distribution of fish collagen peptides using methodology study. To confirm the results, collagen peptides were subjected to ultrafiltration separation and then the filtrates (MW < 3 kDa) were loaded to MALDI-TOF-MS. Superdex column was consider as the most promising candidate for the determination of small molecular weight peptides (MW < 3 kDa), since it spiked recovery was 101.90% and the RSD was below 1%. As for TSK column, it was more suitable for the assay of large molecular weight peptides (MW > 10 kDa). The spiked recovery was 105.66% and the RSD was below 0.5%. Besides, acetonitrile reagent narrowed the pore size distribution of column packing materials, which made it more inaccurate for the analysis of smaller molecular weight peptides.
KeywordsFish collagen peptides Molecular weight distribution (MW distribution) Gel permeation chromatography (GPC) Centrifugal ultrafiltration MALDI-TOF-MS
This study is supported by the National Natural Science Foundation of China [No. 31671804], Guangdong Special Funding for Outstanding Young Scholars [2014TQ01N645], Guangdong Science and Technology Planning Project [2017B090901063 and 2015B020230001], Science and Technology Program of Guangzhou , the Fundamental Research Funds for the Central Universities [017ZD079], and the 111 Project [B17018].
Compliance with Ethical Standards
Conflict of Interest
Xiaoling Lin declares that she has no conflict of interest. Yunjun Lu declares that she has no conflict of interest. Ting Zhang declares that she has no conflict of interest. Ming Liang declares that she has no conflict of interest. Yueyan Cen declares that she has no conflict of interest. Erdong Yuan declares that she has no conflict of interest. Jiaoyan Ren declares that she has no conflict of interest.
This article does not contain any studies with human or animal subjects by any of the authors.
- Haider, S. R., Reid, H. J., & Sharp, B. L. (2012). Tricine-SDS-PAGE. Protein Electrophoresis (pp. 81–91). Springer. https://doi.org/10.1007/978-1-61779-821-4_8
- Kapel R, Klingenberg F, Framboisier X, Dhulster P, Marc I (2011) An original use of size exclusion-HPLC for predicting the performances of batch ultrafiltration implemented to enrich a complex protein hydrolysate in a targeted bioactive peptide. J Membr Sci 383(1–2):26–34. https://doi.org/10.1016/j.memsci.2011.08.025 CrossRefGoogle Scholar
- Liu P, Huang M, Song S, Hayat K, Zhang X, Xia S, Jia C (2012) Sensory characteristics and antioxidant activities of Maillard reaction products from soy protein hydrolysates with different molecular weight distribution. Food Bioprocess Technol 5(5):1775–1789. https://doi.org/10.1007/s11947-010-0440-3 CrossRefGoogle Scholar
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275Google Scholar
- Mori, S., & Barth, H. G. (2013). Size exclusion chromatography. Springer Science & Business MediaGoogle Scholar
- Noman A, Xu Y, AL-Bukhaiti WQ, Abed SM, Ali AH, Ramadhan AH et al (2018) Influence of enzymatic hydrolysis conditions on the degree of hydrolysis and functional properties of protein hydrolysate obtained from Chinese sturgeon (Acipenser sinensis) by using papain enzyme. Process Biochem 67:19–28. https://doi.org/10.1016/j.procbio.2018.01.009 CrossRefGoogle Scholar
- Qin L, Zhu B-W, Zhou D-Y, Wu H-T, Tan H, Yang J-F, Li DM, Dong XP, Murata Y (2011) Preparation and antioxidant activity of enzymatic hydrolysates from purple sea urchin (Strongylocentrotus nudus) gonad. LWT-Food Science and Technology 44(4):1113–1118. https://doi.org/10.1016/j.lwt.2010.10.013 CrossRefGoogle Scholar
- Singh S, Tanwer BS, Khan M (2011) Callus induction and in vivo and in vitro comparative study of primary metabolites of Withania Somnifera. Adv Appl Sci Res 2(3):47–52Google Scholar
- Tometsuka C, Koyama Y-i, Ishijima T, Toyoda T, Teranishi M, Takehana K, Abe K, Nakai Y (2017) Collagen peptide ingestion alters lipid metabolism-related gene expression and the unfolded protein response in mouse liver. Br J Nutr 117:1), 1–1),11. https://doi.org/10.1017/S0007114516004384 CrossRefGoogle Scholar
- Zhang, G., Annan, R. S., Carr, S. A., & Neubert, T. A. (2014). Overview of peptide and protein analysis by mass spectrometry. Current protocols in molecular biology, 10.21. 1–10.21. 30. https://doi.org/10.1002/0471142727.mb1021s108