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Determination and Application of UHPLC-ESI-MS/MS Based Omega Fatty Acids on PUFA Filter Paper with Human Asthma Serum

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Abstract

We developed and validated a fast, sensitive, and accurate analytical method using ultra high-pressure liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI–MS/MS) for simultaneous extraction and quantification of 5 targeted omega fatty acids on human dried serum spots. MS/MS was performed with multiple reaction monitoring (MRM) mode under negative electrospray ionization. We targeted omega fatty acids which are likely to have a positive effect on various inflammatory reactions such as asthma, chronic obstructive pulmonary diseases and rheumatoid arthritis. If applicable, it is necessary to determine them as asthma biomarkers for intervention and public health. The coefficient of determination (r2) of calibration curve exhibited a good linear relationship between 0.9942 and 0.9963 in a range of 0.05–5 µg mL−1. The analytical method was validated with excellent sensitivity (limit of detection; 0.005–0.01 µg mL−1, limit of quantification; 0.03–0.1 µg mL) that allows targeted analyses of omega fatty acids. We obtained the recoveries of 92.6 to 110.4% (RSD, 0.2–9.6%) for intraday and 92.6 to 107.6% (RSD, 1.4–7.0%) for interday. Clinical correlation between the healthy subjects and asthmatic patients’ sera was evaluated with the ratio of omega-3/omega-6 fatty acids. The developed method combines the advantages of easy handling dried filter paper and UHPLC-MS/MS for effectiveness of extraction and analyses step. We evaluated the validation for the quantification and applicability of human serum as a possible role of asthma biomarkers.

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Abbreviations

AA:

Arachidonic acid

ALA:

Alpha-linolenic acid

BSA:

Bovine serum albumin

DBS:

Dried blood spot

DHA:

Docosahexaenoic acid

DPA:

Docosapentaenoic acid

EPA:

Eicosapentaenoic acid

GC–MS:

Gas chromatography–mass spectroscopy

IS:

Internal standards

LLE:

Liquid–liquid extraction

MPA:

Mobile phase A

MPB:

Mobile phase B

MRM:

Multiple reaction monitoring

MTBE:

Methyl tert-butyl ether

PLS-DA:

Partial least square discriminant analysis

PUFAs:

Polyunsaturated fatty acids

SPE:

Solid phase extraction

SPM:

Specialized pro-resolving mediators

UHPLC-ESI-MS/MS:

Ultra high-pressure liquid chromatography-electrospray ionization tandem mass spectrometry

VIP:

Variable importance in the projection

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Funding

This reseach was funded by Duksung Women’s University Research Fund, Seoul, South Korea.

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  1. Biomedical and Pharmaceutical Analysis, College of Pharmacy, Duksung Women’s University, Seoul, South Korea

    Hyejin Lee & Hye-Ran Yoon

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  1. Hyejin Lee
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  2. Hye-Ran Yoon
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Correspondence to Hye-Ran Yoon.

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Lee, H., Yoon, HR. Determination and Application of UHPLC-ESI-MS/MS Based Omega Fatty Acids on PUFA Filter Paper with Human Asthma Serum. Chromatographia 85, 281–294 (2022). https://doi.org/10.1007/s10337-022-04132-4

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