| DOI | Resolve DOI: https://doi.org/10.1016/j.jchromb.2014.12.007 |
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| Author | Search for: Purves, Randy W.1; Search for: Ambrose, Stephen J.1; Search for: Clark, Shawn M.1; Search for: Stout, Jake M.; Search for: Page, Jonathan E.1 |
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| Affiliation | - National Research Council of Canada. Aquatic and Crop Resource Development
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| Format | Text, Article |
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| Subject | acyl coenzyme A; butyryl-coenzyme A; hexanoyl-coenzyme A; isobutyryl-coenzyme A; isovaleryl-coenzyme A; valeryl-coenzyme A; high performance liquid chromatography; isolation and purification; tandem mass spectrometry; Acyl Coenzyme A; Chromatography, High Pressure Liquid; Tandem Mass Spectrometry |
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| Abstract | Acyl coenzyme A (acyl-CoA) thioesters are important intermediates in cellular metabolism and being able to distinguish among them is critical to fully understanding metabolic pathways in plants. Although significant advances have been made in the identification and quantification of acyl-CoAs using liquid chromatography tandem mass spectrometry (LC-MS/MS), separation of isomeric species such as isobutyryl- and n-butyrl-CoA has remained elusive. Here we report an ultra-performance liquid chromatography (UPLC)-MS/MS method for quantifying short-chain acyl-CoAs including isomeric species n-butyryl-CoA and isobutyryl-CoA as well as n-valeryl-CoA and isovaleryl-CoA. The method was applied to the analysis of extracts of hop (Humulus lupulus) and provided strong evidence for the existence of an additional structural isomer of valeryl-CoA, 2-methylbutyryl-CoA, as well as an unexpected isomer of hexanoyl-CoA. The results showed differences in the acyl-CoA composition among varieties of Humulus lupulus, both in glandular trichomes and cone tissues. When compared with the analysis of hemp (Cannabis sativa) extracts, the contribution of isobutyryl-CoAs in hop was greater as would be expected based on the downstream polyketide products. Surprisingly, branched chain valeryl-CoAs (isovaleryl-CoA and 2-methylbutyryl-CoA) were the dominant form of valeryl-CoAs in both hop and hemp. The capability to separate these isomeric forms will help to understand biochemical pathways leading to specialized metabolites in plants. |
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| Publication date | 2015-02-01 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 21277028 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | d9590485-6223-4f21-b29f-6e2558ad6e4d |
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| Record created | 2015-11-10 |
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| Record modified | 2020-04-22 |
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