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Determination of Triacylglycerols in Butterfat by Normal-Phase HPLC and Electrospray–Tandem Mass Spectrometry

  • Original Article
  • Published:
Lipids

Abstract

Here, we report the identification and quantification of the molecular species of long-, medium-, and short-chain triacylglycerols (TAG) in butterfat (BF), including TAG with an odd number of acyl carbons (ACN) and TAG with unidentified molecular species. In the present study, in addition to auto-MS2, a large number of methods, each recording MS2 for 1–4 ions, were used for identification of TAG species. For the quantification of long-chain, odd ACN TAG, and TAG with unidentified molecular species, molar correction factors (MCF) were calculated from the uncorrected mol% (area mol%) of each ACN:DB (number of double bonds) class in randomized butterfat (RBF), and the respective mol% in the calculated random composition of RBF. The butyrate, caproate, and medium-chain (C8, C10) TAG were quantified using regio- or acyl-chain-specific MCF calculated from their area mol% in RBF and mol% in the calculated random composition. These methods enabled us to identify ca. 450 TAG species in 184 quantified peaks of 88 ACN:DB classes. The proportions of saturated, monoene, diene, triene, tetraene, pentaene, and hexaene TAG were 40.0, 38.4, 16.2, 4.5, 0.6, 0.1, and 0.03 mol%, respectively. The proportions of TAG with not identified molecular species and odd ACN TAG were 11.8 and 5.7 mol%, respectively. The most abundant short-chain TAG species were butyroyldipalmitoylglycerol + butyroylmyristoylstearoylglycerol (5.25 mol%) and butyroylpalmitoyloleoylglycerol (4.08 mol%).

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Abbreviations

ACN:

The number of acyl group carbons

ACN:DB:

The number of acyl group carbons:the number of double bonds

APCI:

Atmospheric pressure chemical ionization

BF:

Butterfat

CI:

Chemical ionization

DAG:

Diacylglycerol(s)

EI–MS:

Electron ionization mass spectrometry

ESI–MS:

Electrospray mass spectrometry

ESI–MS2 :

Electrospray tandem mass spectrometry

GC:

Gas–liquid chromatography

HPLC:

High performance liquid chromatography

ISTD:

Internal standard

LC–MS:

Liquid chromatography–mass spectrometry

MCF:

Molar correction factor

MF:

Milk fat

NP:

Normal-phase

RBF:

Randomized butterfat

RP:

Reversed-phase

RRT:

Relative retention time

SFC:

Supercritical fluid chromatography

sn :

Stereospecific numbering

SPE:

Solid phase extraction

TAG:

Triacylglycerol(s)

TLC:

Thin layer chromatography

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Acknowledgments

Financial support from the Jenny and Antti Wihuri Foundation and Finnish Society of Dairy Science is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Applied Chemistry and Microbiology, University of Helsinki, Helsinki, Finland

    P. Kalo & V. Ollilainen

  2. Department of Food Technology, University of Helsinki, Helsinki, Finland

    A. Kemppinen

Authors
  1. P. Kalo
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  2. A. Kemppinen
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  3. V. Ollilainen
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Corresponding author

Correspondence to P. Kalo.

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Kalo, P., Kemppinen, A. & Ollilainen, V. Determination of Triacylglycerols in Butterfat by Normal-Phase HPLC and Electrospray–Tandem Mass Spectrometry. Lipids 44, 169–195 (2009). https://doi.org/10.1007/s11745-008-3247-5

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  • DOI: https://doi.org/10.1007/s11745-008-3247-5

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