Isolation of α-linolenic acid biohydrogenation products by combined silver ion solid phase extraction and semi-preparative high performance liquid chromatography
Turner, TD; Meadus, WJ; Mapiye, C; Vahmani, P; López-Campos, Ó; Duff, P; Rolland, DC; Church, JS; Dugan, ME
HERO ID
2902265
Reference Type
Journal Article
Year
2015
Language
English
PMID
| HERO ID | 2902265 |
|---|---|
| In Press | No |
| Year | 2015 |
| Title | Isolation of α-linolenic acid biohydrogenation products by combined silver ion solid phase extraction and semi-preparative high performance liquid chromatography |
| Authors | Turner, TD; Meadus, WJ; Mapiye, C; Vahmani, P; López-Campos, Ó; Duff, P; Rolland, DC; Church, JS; Dugan, ME |
| Journal | Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences |
| Volume | 980 |
| Page Numbers | 34-40 |
| Abstract | Polyunsaturated fatty acids typically found in cattle feed include linoleic (LA) and α-linolenic acid (ALA). In the rumen, microbes metabolize these resulting in the formation of biohydrogenation products (BHP), which can be incorporated into meat and milk. Bioactivities of LA-BHP, including conjugated linoleic acid (cis (c) 9,trans (t) 11-18:2 and t10,c12-18:2) and trans fatty acid isomers (t9-, t10- and t11-18:1) have been investigated, but effects of several BHP unique to ALA have not been extensively studied, and most ALA-BHP are not commercially available. The objective of the present research was to develop methods to purify and collect ALA-BHP using silver ion (Ag(+)) chromatography in sufficient quantities to allow for convenient bioactivity testing in cell culture. Fatty acid methyl esters (FAME) were prepared from perirenal adipose tissue from a cow enriched with ALA-BHP by feeding flaxseed. These were applied to Ag(+)-solid phase extraction, and eluted with hexane with increasing quantities of acetone (1, 2, 10, 20%) or acetonitrile (2%) to pre-fractionate FAME based on degree of unsaturation and double bond configuration. Fractions were collected, concentrated and applied to semi-preparative Ag(+)-high performance liquid chromatography (HPLC) for the isolation and collection of purified isomers, which was accomplished using isocratic elutions with hexane containing differing amounts of acetonitrile (from 0.015 to 0.075%). Purified trans-18:1 isomers collected ranged in purity from 88 to 99%. Purity of the ALA-BHP dienes collected, including c9,t13-18:2, t11,c15-18:2 and t10,c15-18:2, exceeded 90%, while purification of other dienes may require the use of other complementary procedures (e.g. reverse phase HPLC). |
| Doi | 10.1016/j.jchromb.2014.11.038 |
| Pmid | 25579113 |
| Wosid | WOS:000348889500006 |
| Is Certified Translation | No |
| Dupe Override | No |
| Comments | Scopus URL: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84920913660&doi=10.1016%2fj.jchromb.2014.11.038&partnerID=40&md5=457c0fffcaa907cadb305c3407cfdb7f |
| Is Public | Yes |
| Language Text | English |
| Keyword | Ag+-HPLC; Ag+-SPE; Biohydrogenation; α-Linolenic acid |