A practical and fast isolation of 12 cyclolinopeptides (linusorbs) from flaxseed oil via preparative HPLC with phenyl-hexyl column

Xue Liu; Zi-Zhe Cai; Wan Jun Lee; Xuan-Xuan Lu; Martin J T Reaney; Jian-Ping Zhang; Ying Li; Ning Zhang; Yong Wang

doi:10.1016/j.foodchem.2021.129318

A practical and fast isolation of 12 cyclolinopeptides (linusorbs) from flaxseed oil via preparative HPLC with phenyl-hexyl column

Food Chem. 2021 Jul 30:351:129318. doi: 10.1016/j.foodchem.2021.129318. Epub 2021 Feb 18.

Authors

Xue Liu¹, Zi-Zhe Cai¹, Wan Jun Lee¹, Xuan-Xuan Lu¹, Martin J T Reaney², Jian-Ping Zhang³, Ying Li¹, Ning Zhang⁴, Yong Wang⁵

Affiliations

¹ Guangdong Saskatchewan Oilseed Joint Laboratory, Department of Food Science and Engineering, Jinan University, Guangzhou 510632, China.
² Guangdong Saskatchewan Oilseed Joint Laboratory, Department of Food Science and Engineering, Jinan University, Guangzhou 510632, China; Department of Plant Sciences, University of Saskatchewan, 51 Campus Dr., Saskatoon, SK, S7N 5A8, Canada.
³ Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China.
⁴ Guangdong Saskatchewan Oilseed Joint Laboratory, Department of Food Science and Engineering, Jinan University, Guangzhou 510632, China. Electronic address: tzhning@jnu.edu.cn.
⁵ Guangdong Saskatchewan Oilseed Joint Laboratory, Department of Food Science and Engineering, Jinan University, Guangzhou 510632, China. Electronic address: twyong@jnu.edu.cn.

PMID: 33647690
DOI: 10.1016/j.foodchem.2021.129318

Abstract

Linusorbs, known as cyclolinopeptides, are a group of cyclic hydrophobic peptides derived from flaxseed oil with various health benefits. However, the current research efforts on both the biological activities and antioxidant capacities of linusorbs are limited because of existing issues with their purification and characterization. A practical method based on preparative HPLC for isolating 12 linusorbs simultaneously was developed and factors such as the solvent selection, gradient elution program, flow rate, loaded mass, and loading concentration, were optimized. The optimum conditions were an initial acetonitrile (ACN) to water ratio of 40%, final ACN ratio of 80%, eluting time of 21 min, a flow rate of 16 mL/min, sample load of 12.5 mg, and concentration of 80 mg/mL (in methanol). The 12 linusorbs obtained were verified using off-line MS/MS, recording purities of above 95.5%. The method could serve as a practical and fast isolation method enabling further investigation of minor linusorbs.

Keywords: Cyclolinopeptide; Flaxseed oil; Phenyl-hexyl column; Preparative HPLC.

MeSH terms

Acetonitriles / chemistry
Chromatography, High Pressure Liquid / methods*
Hydrophobic and Hydrophilic Interactions
Linseed Oil / chemistry*
Methanol / chemistry
Peptides, Cyclic / chemistry
Peptides, Cyclic / isolation & purification*
Time Factors

Substances

Acetonitriles
Peptides, Cyclic
Linseed Oil
Methanol
acetonitrile