Triglyceride quantification by catalytic saturation and LC-MS/MS reveals an evolutionary divergence in regioisometry among green microalgae

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A method for the specific quantification of triacylglycerides (TGs) using electrospray ionization (ESI) reverse-phase liquid chromatography tandem mass spectroscopy (LC-MS/MS) with multiple reaction monitoring (MRM) was developed for routine high throughput applications. Incorporation of catalytic hydrogenation dramatically reduced TG pool complexity, enabling the absolute quantification of TG and regioisomers at the expense of double bond frequency and placement. Saturated TAG standards demonstrated equivalent linear detector responses to increases in concentration over two orders of magnitude. Accuracy was evaluated against a standard gas chromatography mass spectroscopy (GC-MS) protocol using three plant-derived oils and biological lipid extracts of two algal genera, Chlamydomonas reinhardtii and Coccomyxa subellipsoidea. Quantification of TG in palm and olive oils was > 91% accurate compared to the GC-MS method, and the content of TG molecular species was comparable to literature values. Differences in algal TG quantities were statistically insignificant. The ratio of the product ion peak areas for pairs of regioisomers was linearly correlated to their relative abundance. Quantification of regioisomers enabled sn-2 positional analyses related to the amount of TG synthesized in the chloroplast of photosynthetic organisms. A demonstrative survey of 35 algae revealed an evolutionary divergence of the dominant TG synthetic pathways by order Chlamydomnadales.

Original languageEnglish (US)
Pages (from-to)23-31
Number of pages9
JournalAlgal Research
Volume5
Issue number1
DOIs
StatePublished - Jan 1 2014

Fingerprint

divergent evolution
microalgae
triacylglycerols
mass spectrometry
algae
Coccomyxa
gas chromatography
hydrogenation
reversed-phase liquid chromatography
Chlamydomonas reinhardtii
palm oils
autotrophs
olive oil
ionization
detectors
chloroplasts
ions
oils
monitoring
extracts

Keywords

  • Biofuels
  • LC-MS/MS
  • Prokaryotic pathway
  • Regioisometry
  • Stereospecificity
  • Triglycerides

ASJC Scopus subject areas

  • Agronomy and Crop Science

Cite this

@article{436c338f6af94189ac9f03e670e46127,
title = "Triglyceride quantification by catalytic saturation and LC-MS/MS reveals an evolutionary divergence in regioisometry among green microalgae",
abstract = "A method for the specific quantification of triacylglycerides (TGs) using electrospray ionization (ESI) reverse-phase liquid chromatography tandem mass spectroscopy (LC-MS/MS) with multiple reaction monitoring (MRM) was developed for routine high throughput applications. Incorporation of catalytic hydrogenation dramatically reduced TG pool complexity, enabling the absolute quantification of TG and regioisomers at the expense of double bond frequency and placement. Saturated TAG standards demonstrated equivalent linear detector responses to increases in concentration over two orders of magnitude. Accuracy was evaluated against a standard gas chromatography mass spectroscopy (GC-MS) protocol using three plant-derived oils and biological lipid extracts of two algal genera, Chlamydomonas reinhardtii and Coccomyxa subellipsoidea. Quantification of TG in palm and olive oils was > 91{\%} accurate compared to the GC-MS method, and the content of TG molecular species was comparable to literature values. Differences in algal TG quantities were statistically insignificant. The ratio of the product ion peak areas for pairs of regioisomers was linearly correlated to their relative abundance. Quantification of regioisomers enabled sn-2 positional analyses related to the amount of TG synthesized in the chloroplast of photosynthetic organisms. A demonstrative survey of 35 algae revealed an evolutionary divergence of the dominant TG synthetic pathways by order Chlamydomnadales.",
keywords = "Biofuels, LC-MS/MS, Prokaryotic pathway, Regioisometry, Stereospecificity, Triglycerides",
author = "Allen, {James W.} and DiRusso, {Concetta C} and Black, {Paul N}",
year = "2014",
month = "1",
day = "1",
doi = "10.1016/j.algal.2014.04.003",
language = "English (US)",
volume = "5",
pages = "23--31",
journal = "Algal Research",
issn = "2211-9264",
publisher = "Elsevier BV",
number = "1",

}

TY - JOUR

T1 - Triglyceride quantification by catalytic saturation and LC-MS/MS reveals an evolutionary divergence in regioisometry among green microalgae

AU - Allen, James W.

AU - DiRusso, Concetta C

AU - Black, Paul N

PY - 2014/1/1

Y1 - 2014/1/1

N2 - A method for the specific quantification of triacylglycerides (TGs) using electrospray ionization (ESI) reverse-phase liquid chromatography tandem mass spectroscopy (LC-MS/MS) with multiple reaction monitoring (MRM) was developed for routine high throughput applications. Incorporation of catalytic hydrogenation dramatically reduced TG pool complexity, enabling the absolute quantification of TG and regioisomers at the expense of double bond frequency and placement. Saturated TAG standards demonstrated equivalent linear detector responses to increases in concentration over two orders of magnitude. Accuracy was evaluated against a standard gas chromatography mass spectroscopy (GC-MS) protocol using three plant-derived oils and biological lipid extracts of two algal genera, Chlamydomonas reinhardtii and Coccomyxa subellipsoidea. Quantification of TG in palm and olive oils was > 91% accurate compared to the GC-MS method, and the content of TG molecular species was comparable to literature values. Differences in algal TG quantities were statistically insignificant. The ratio of the product ion peak areas for pairs of regioisomers was linearly correlated to their relative abundance. Quantification of regioisomers enabled sn-2 positional analyses related to the amount of TG synthesized in the chloroplast of photosynthetic organisms. A demonstrative survey of 35 algae revealed an evolutionary divergence of the dominant TG synthetic pathways by order Chlamydomnadales.

AB - A method for the specific quantification of triacylglycerides (TGs) using electrospray ionization (ESI) reverse-phase liquid chromatography tandem mass spectroscopy (LC-MS/MS) with multiple reaction monitoring (MRM) was developed for routine high throughput applications. Incorporation of catalytic hydrogenation dramatically reduced TG pool complexity, enabling the absolute quantification of TG and regioisomers at the expense of double bond frequency and placement. Saturated TAG standards demonstrated equivalent linear detector responses to increases in concentration over two orders of magnitude. Accuracy was evaluated against a standard gas chromatography mass spectroscopy (GC-MS) protocol using three plant-derived oils and biological lipid extracts of two algal genera, Chlamydomonas reinhardtii and Coccomyxa subellipsoidea. Quantification of TG in palm and olive oils was > 91% accurate compared to the GC-MS method, and the content of TG molecular species was comparable to literature values. Differences in algal TG quantities were statistically insignificant. The ratio of the product ion peak areas for pairs of regioisomers was linearly correlated to their relative abundance. Quantification of regioisomers enabled sn-2 positional analyses related to the amount of TG synthesized in the chloroplast of photosynthetic organisms. A demonstrative survey of 35 algae revealed an evolutionary divergence of the dominant TG synthetic pathways by order Chlamydomnadales.

KW - Biofuels

KW - LC-MS/MS

KW - Prokaryotic pathway

KW - Regioisometry

KW - Stereospecificity

KW - Triglycerides

UR - http://www.scopus.com/inward/record.url?scp=84900956854&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84900956854&partnerID=8YFLogxK

U2 - 10.1016/j.algal.2014.04.003

DO - 10.1016/j.algal.2014.04.003

M3 - Article

AN - SCOPUS:84900956854

VL - 5

SP - 23

EP - 31

JO - Algal Research

JF - Algal Research

SN - 2211-9264

IS - 1

ER -