Small volume retinol binding protein measurement by liquid chromatography-tandem mass spectrometry

William S Phipps; Dina N Greene; Hannah Pflaum; Thomas J Laha; Jane A Dickerson; Jill Irvine; Anna E Merrill; Pratistha Ranjitkar; Clark M Henderson; Andrew N Hoofnagle

doi:10.1016/j.clinbiochem.2021.10.005

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Small volume retinol binding protein measurement by liquid chromatography-tandem mass spectrometry

Journal article

Peer reviewed

Small volume retinol binding protein measurement by liquid chromatography-tandem mass spectrometry

William S Phipps, Dina N Greene, Hannah Pflaum, Thomas J Laha, Jane A Dickerson, Jill Irvine, Anna E Merrill, Pratistha Ranjitkar, Clark M Henderson and Andrew N Hoofnagle

Clinical biochemistry, Vol.99, pp.111-117

01/2022

DOI: 10.1016/j.clinbiochem.2021.10.005

PMCID: PMC8671195

PMID: 34678307

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https://www.ncbi.nlm.nih.gov/pmc/articles/8671195View

Open Access

Abstract

The measurement of plasma concentrations of retinol binding protein is a component of nutritional assessment in neonatal intensive care. However, serial testing in newborns is hampered by the limited amount of blood that can be sampled. Limitations are most severe with preterm infants, for whom close monitoring may be most important. We developed an assay to quantify retinol binding protein using trypsin digestion and liquid chromatography-tandem mass spectrometry, which requires a serum or plasma volume of 5 µl. Additionally, we validated the method according to current recommendations and performed comparison with a standard nephelometry platform in clinical use. The assay demonstrated linearity from below 1 mg/dL (0.48 µM) to more than 20 mg/dL (9.7 µM), and an imprecision of 11.8% at 0.43 mg/dL (0.21 µM). The distribution of results observed with the new method was different when compared with nephelometry. Liquid chromatography-tandem mass spectrometry facilitated testing a smaller sample volume, thereby increasing the ability to monitor key nutritional markers in premature infants. The differences in results compared with a commercially-available nephelometric assay revealed questionable results for lower concentrations by immunoassay.

LC-MS/MS

Liquid chromatography-tandem mass spectrometry

Nutrition

Retinol binding protein

Vitamin A

Details

Title: Subtitle: Small volume retinol binding protein measurement by liquid chromatography-tandem mass spectrometry
Creators: William S Phipps - University of Washington
Dina N Greene - University of Washington
Hannah Pflaum - University of Washington
Thomas J Laha - University of Washington
Jane A Dickerson - University of Washington
Jill Irvine - University of Washington Medical Center
Anna E Merrill - University of Washington
Pratistha Ranjitkar - University of Washington
Clark M Henderson - University of Washington
Andrew N Hoofnagle - University of Washington
Resource Type: Journal article
Publication Details: Clinical biochemistry, Vol.99, pp.111-117
DOI: 10.1016/j.clinbiochem.2021.10.005
PMID: 34678307
PMCID: PMC8671195
NLM abbreviation: Clin Biochem
ISSN: 0009-9120
eISSN: 1873-2933
Publisher: Elsevier Inc
Language: English
Date published: 01/2022
Academic Unit: Pathology; Injury Prevention Research Center
Record Identifier: 9984201421102771

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