Non-aqueous reversed phase liquid chromatography with charged aerosol detection for quantitative lipid analysis with improved accuracy
Causevic, Ariana LU ; Olofsson, Kim LU ; Adlercreutz, Patrick LU
and Grey, Carl
LU
(2021)
In Journal of Chromatography A
1652.
- Abstract
There is a great need for efficient analysis of the composition of vegetable oils and fats, since it affects the physical and technical properties. However, due to the complex nature of these kind of samples, it is often difficult and costly. In the present study, we developed a Non-Aqueous Reversed-Phase HPLC method that can be used to separate and quantify different free fatty acids, fatty acid esters, monoacylglycerides, diacylglycerides and triacylglycerides, including regioisomers such as SOS/SSO and 1,2- and 1,3-diolein. Two 25 cm Nucleodur C18 Isis columns in series, sub-ambient column temperature and a mobile phase gradient composed of acetonitrile, acetic acid, isopropanol and heptane were used for the separation. The lipids... (More)
There is a great need for efficient analysis of the composition of vegetable oils and fats, since it affects the physical and technical properties. However, due to the complex nature of these kind of samples, it is often difficult and costly. In the present study, we developed a Non-Aqueous Reversed-Phase HPLC method that can be used to separate and quantify different free fatty acids, fatty acid esters, monoacylglycerides, diacylglycerides and triacylglycerides, including regioisomers such as SOS/SSO and 1,2- and 1,3-diolein. Two 25 cm Nucleodur C18 Isis columns in series, sub-ambient column temperature and a mobile phase gradient composed of acetonitrile, acetic acid, isopropanol and heptane were used for the separation. The lipids were detected and quantified using a charged aerosol detector and it was found that the peak shape highly affected the detector response as well as the response uniformity, even when inverse gradient compensation was employed. Thus, calibration and determination of response factors were necessary for reliable quantification. A correlation between response factors and peak width at half peak height was found and used for quantification of non-calibrated components. A quantification approach was suggested including an appropriate selection of calibrated components, depending on sample composition and the accuracy required. It was shown in a complex oil sample that the reduced calibration approach, using only 6 instead of 33 calibrated components, resulted in virtually the same composition, but yielded a more accurate result compared to using relative area that neglects response factors. The method validation showed good reproducibility and accuracy, making it an excellent tool for extensive analysis of complex lipid mixtures.
(Less)
- author
- Causevic, Ariana
LU
; Olofsson, Kim
LU
; Adlercreutz, Patrick
LU
and Grey, Carl
LU
- organization
- publishing date
- 2021-08-30
- type
- Contribution to journal
- publication status
- published
- keywords
- CAD, NARP-HPLC, Quantification, Regioisomers, Separation of lipids, Sub-ambient temperature
- in
- Journal of Chromatography A
- volume
- 1652
- article number
- 462374
- publisher
- Elsevier
- external identifiers
-
- pmid:34246965
- scopus:85109458339
- ISSN
- 0021-9673
- DOI
- 10.1016/j.chroma.2021.462374
- language
- English
- LU publication?
- yes
- additional info
- Funding Information: This work was supported by the Swedish Foundation for Strategic Research [grant number ID16-0051 ] and AAK AB.
- id
- b50b7d25-f4f8-40a0-a2e9-05082efd0d2c
- date added to LUP
- 2021-08-17 16:25:40
- date last changed
- 2025-04-04 02:33:13
@article{b50b7d25-f4f8-40a0-a2e9-05082efd0d2c,
abstract = {{<p>There is a great need for efficient analysis of the composition of vegetable oils and fats, since it affects the physical and technical properties. However, due to the complex nature of these kind of samples, it is often difficult and costly. In the present study, we developed a Non-Aqueous Reversed-Phase HPLC method that can be used to separate and quantify different free fatty acids, fatty acid esters, monoacylglycerides, diacylglycerides and triacylglycerides, including regioisomers such as SOS/SSO and 1,2- and 1,3-diolein. Two 25 cm Nucleodur C18 Isis columns in series, sub-ambient column temperature and a mobile phase gradient composed of acetonitrile, acetic acid, isopropanol and heptane were used for the separation. The lipids were detected and quantified using a charged aerosol detector and it was found that the peak shape highly affected the detector response as well as the response uniformity, even when inverse gradient compensation was employed. Thus, calibration and determination of response factors were necessary for reliable quantification. A correlation between response factors and peak width at half peak height was found and used for quantification of non-calibrated components. A quantification approach was suggested including an appropriate selection of calibrated components, depending on sample composition and the accuracy required. It was shown in a complex oil sample that the reduced calibration approach, using only 6 instead of 33 calibrated components, resulted in virtually the same composition, but yielded a more accurate result compared to using relative area that neglects response factors. The method validation showed good reproducibility and accuracy, making it an excellent tool for extensive analysis of complex lipid mixtures.</p>}},
author = {{Causevic, Ariana and Olofsson, Kim and Adlercreutz, Patrick and Grey, Carl}},
issn = {{0021-9673}},
keywords = {{CAD; NARP-HPLC; Quantification; Regioisomers; Separation of lipids; Sub-ambient temperature}},
language = {{eng}},
month = {{08}},
publisher = {{Elsevier}},
series = {{Journal of Chromatography A}},
title = {{Non-aqueous reversed phase liquid chromatography with charged aerosol detection for quantitative lipid analysis with improved accuracy}},
url = {{http://dx.doi.org/10.1016/j.chroma.2021.462374}},
doi = {{10.1016/j.chroma.2021.462374}},
volume = {{1652}},
year = {{2021}},
}