Details:

Alternative Title:
J Appl Lab Med
Personal Author:
Chaudhary-Webb, Madhulika ; Schleicher, Rosemary L. ; Erhardt, Juergen G. ; Pendergrast, Elizabeth C. ; Pfeiffer, Christine M.
Chaudhary-Webb, Madhulika ; Schleicher, Rosemary L. ; Erhardt, Juergen G. ; Pendergrast, Elizabeth C. ; Pfeiffer, Christine M. Less -
Description:
Background:

Assessing vitamin A status in populations remains a high public health priority for low- and middle-income countries. However, analytical difficulties with serum retinol measurements persist in international laboratories. Nearly all participants in a Centers for Disease Control and Prevention external quality assessment program use HPLC to measure serum retinol, but round-to-round results failing to meet acceptable criteria suggest the need to provide a straightforward stable HPLC ultraviolet (UV) method that can be adopted by these laboratories to improve performance. We present a protein precipitation HPLC-UV method that measures serum retinol below the deficiency cutoff value (<0.7 μmol/L or 20 μg/dL) that is suitable for low- and middle-income countries and uses commercially available materials.

Methods:

Serum (25 μL) added to retinyl acetate was precipitated with acetonitrile (125 μL) to extract retinol. Solvent-based calibration solutions required no extraction. Calibration used either single-point (50 μg/dL) or multipoint solutions (0.52–100 μg/dL). C18 column (4.6 × 100 mm) and acetonitrile with 0.1% triethylamine/water (83/17, v/v) as isocratic mobile phase (1.1 mL/min), achieved baseline separation (7 minutes).

Results:

With only 25 μL of serum, the limit of detection was 0.52 μg/dL. Single- and multipoint calibration generated equivalent results. Over several years, between-run imprecision was ≤7.1% in multiple quality-control materials. Overall mean (CV) method bias for NIST-certified reference materials (e-series) was −0.2% (5.8%). Maximally, 180 samples were processed within 24 h.

Conclusions:

This method was robust and stable over years and accurately measured serum retinol with low-volume samples. Thus, it may be of interest to low- and middle-income countries and to pediatric and finger stick applications.


Subjects:
[+]
Article
Source:
J Appl Lab Med. 4(1):101-107
Pubmed ID:
31639712
Pubmed Central ID:
PMC6945745
Document Type:
Journal Article
Funding:
CC999999/ImCDC/Intramural CDC HHS/United States
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:57f85455759889590a34bdd9945c931cbd46aa63a7228177a64b5ad7be014f49
Download URL:
https://stacks.cdc.gov/view/cdc/84059/cdc_84059_DS1.pdf
File Type:

Supporting Files

• 	nihms-1063967-f0001.gif	gif
  	nihms-1063967-f0001.jpg	jpeg
  	nihms-1063967.nxml	xml
  	NIHMS1063967-supplement-Supp_Fig_1.pdf	pdf
  	NIHMS1063967-supplement-Supp_Fig_2.pdf	pdf

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