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. 2021 May 18;13(5):1703. doi: 10.3390/nu13051703

Table 2.

Summary of methods for determination of retinoids and carotenoids in human biological materials.

Technique Sensitivity (nmol/L) Matrix Analytes Advantages Disadvantages References
HPLC-UV-vis/DAD 1 0.1 × 10−3–209.46 Serum, plasma, seminal plasma, mouse embryos and kidney, dried whole blood spots, breast milk, red blood cells, adipose tissue Retinoids (retinol, retinal, RA, retinyl esters) and carotenoids Usually small sample volume (30–200 µL)
Combination with automation and online sample preparation
Short analysis times in some multicomponent analyses
Some methods have comparable sensitivity to MS detection
Long analysis time with a complicated gradient
Use of a large volume of toxic solvents as the mobile phase and in sample preparation procedures, mainly in carotenoids analyses
[279,280,281,282,283,284,285,286,287,288,289,290,291,292,293,294,295]
HPLC-FLD 1 2.3–34.91 Plasma, breast milk Retinol Not often used
No sufficient data
[296,297]
LC–MS
LC–MS/MS
2 2 × 10−6–261.83 Urine, plasma, amniotic fluid, tears, serum Retinol, RA Usually, simple sample preparation procedures In the case of retinoic acid usage of large volumes of toxic solvents in sample preparation procedures with poor recovery [298,299,300,301,302,303]
SFC-MS/MS 1 0.09 × 10−6–70.31 Whole blood, plasma, serum, colostrum Carotenoids, apocarotenoids, epoxycarotenoids Small sample volume (10–200 µL)
Short analysis times in multicomponent analyses
Combination with online SFE or robotic SLE
[304,305,306,307]
HPLC-ECD 1 0.4 × 10−3–314.19 Serum, rat plasma, cervical tissue Retinol, RA, carotenoids Small sample volume (20–200 µL),
Short analysis times in multicomponent analyses of retinol and RA
Long analysis time of carotenoids
Toxic solvents usage
[308,309,310]
ELISA kits 1 0.11–279.38 Whole blood, serum, cell lysates, plasma, tissues, other human liquids, colostrum Retinol, β-carotene One kit for various matrices (serum, plasma, other biofluids, cell lysates) For research only
Cross-reactivity with retinol analogs
Time and money consuming for small sample series
[311,312,313,314,315,316,317,318,319]
HPLC/UHPLC-UV kits 2 23.62–1174.5 Plasma, serum Retinol, carotenoids Small sample volume (50–250 µL)
Some kits: possible to combine with a 96-well plate format
96 samples in 30 min
Some kits: available in UHPLC mode–3 analytes in 3.5 min
Some kits have long analysis times and use large sample volumes (400 µL)
Only for serum or plasma
[320,321,322,323,324]

1 LOD—limit of detection, 2 LLOQ—lower limit of quantification. DAD—diode array detection; ECD—electrochemical detection; ELISA enzyme-linked immunosorbent assay; FLD—fluorescence detection; HPLC—high-performance liquid chromatography; LC–MS—coupling of liquid chromatography and mass spectrometry; RA—retinoic acid; SFC-MS—coupling of supercritical fluid chromatography and mass spectrometry; SFE—supercritical fluid extraction; SLE—solid-supported liquid–liquid extraction; UHPLC—ultra-high-performance liquid chromatography; UV-vis—ultraviolet/visible detection.