High-performance liquid chromatography of the unsaponifiable from samples of marine and freshwater fish: fractionation and identification of retinol (vitamin A1) and dehydroretinol (vitamin A2) isomers

Stancher, B.; Zonta, F

doi:10.1016/s0021-9673(01)87711-6

Cited by 34 publications

(11 citation statements)

References 20 publications

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“…Schmidt et al also provide evidence of an analytically rigorous assay appropriate for tissue retinoid quantification (44), whereas other assays have only been assessed for simpler matrices, such as serum (51,52,55), and/or have not been comprehensively characterized for analytical performance (16,17,19,(22)(23)(24)(25)30,31,36,54). Simplification of the sample matrix also enhances accuracy and the lives of guard and analytical columns.…”

Section: Comparison To Other Methodsmentioning

confidence: 99%

HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues

Kane

Folias

Napoli

2008

Analytical Biochemistry

158

166

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We report robust HPLC/UV methods for quantifying retinyl esters (RE), retinol (ROL) and retinal (RAL) applicable to diverse biological samples, with lower limits of detection of 0.7 pmol, 0.2 pmol, and 0.2 pmol, respectively, and linear ranges >3 orders of magnitude. These assays function well with small, complex biological samples (10-20 mg tissue). Coefficients of variation range from: intra-day, 5.9-10.0%; inter-day, 5.9-11.0%. Quantification of endogenous RE, ROL, and RAL in mouse serum and tissues (liver, kidney, adipose, muscle, spleen, testis, skin, brain, and brain regions) reveals utility. Ability to discriminate spatial concentrations of ROL and RE is illustrated with C57BL/6 mouse brain loci (hippocampus, cortex, olfactory bulb, thalamus, cerebellum, and striatum.) We also developed a method to distinguish isomeric forms of ROL to investigate precursors of retinoic acid. The ROL isomer assay has limits of detection between 3.5-4.5 pmol and a similar linear range and % CV as the ROL/RE and RAL assays. The assays described here provide for sensitive and rigorous quantification of endogenous RE, ROL, and RAL to elucidate retinoid homeostasis in disease states, such as Alzheimer's disease, type 2 diabetes, obesity, and cancer.

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Section: Comparison To Other Methodsmentioning

confidence: 99%

HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues

Kane

Folias

Napoli

2008

Analytical Biochemistry

158

166

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“…To liberate esterified vitamins, homogenates of liver and lipid samples extracted from the adipose tissues with chloroform‐methanol (2:1 v/v) were saponified in an ethanolic KOH solution in the presence of ascorbic acid and under nitrogen. The saponification and vitamin extraction method, described earlier in detail [20], was a modification of the method of Stancher and Zonta [22]. Each plasma sample was prepared by a saponification procedure, and a duplicate was made by direct extraction [23].…”

Section: Methodsmentioning

confidence: 99%

Vitamins A₁, A₂, and E in minks exposed to polychlorinated biphenyls (Aroclor 1242®) and copper, VIA diet based on freshwater or marine fish

Käkelä

Hyvärinen

et al. 1999

Enviro Toxic and Chemistry

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Minks (Mustela vison) fed diets based on either freshwater fish or marine fish were exposed to 1 mg of polychlorinated biphenyls (PCBs) (Aroclor 1242) daily for 28 d. To minks on the freshwater diet, copper (62 mg/kg food) was also given with or without PCBs. The marine diet (vitamin-rich plus additional supplements) included more vitamin A 1 and E than the freshwater diet. We studied how the exposures affected levels of vitamins A 1 , A 2 , and E in liver and adipose tissues and levels of vitamins A 1 and A 2 in plasma. In females and males on the freshwater diet, the hepatic level of vitamin A 2 was decreased because of the PCBs, and in these males the hepatic levels of vitamin E also decreased. Interestingly, with coexposure to PCBs and copper, the vitamin levels were, in general, close to the control values. In adipose tissues also, the PCBs induced significant changes in the concentrations of vitamins A 1 and A 2 . In plasma, vitamins A 1 and A 2 were decreased in all patterns of exposure and on both diets. However, plasma thyroxine was slightly increased, a finding opposite to that reported previously in rodent studies. The results suggest that, in mink, diet greatly modulates the responses to PCBs, which may also differ in males and females. Furthermore, vitamins A 1 and A 2 may not be metabolized equally during PCB administration. Keywords-Polychlorinated biphenyls Vitamin A Vitamin E Mink CopperEnviron. Toxicol. Chem. 18, 1999 R. Käkelä et al.

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“…The retention times (relative to A 1 ‐16:0) of the A 2 esters were 0.848 ± 0.003 times those of the corresponding A 1 esters in 100% MeOH and 0.820 ± 0.003 in the argentated methanol, respectively. Because of the different absorption spectra and maxima of A 1 (325 nm) and A 2 (350 nm) [22,23], the esters of the two analogs can be separated when detected at both wavelengths. As far as we know, the retention properties of the numerous A 2 esters have not been reported previously.…”

Section: Resultsmentioning

confidence: 99%

“…In the otters about one‐half and in the mink about one‐fourth of the vitamin A totals were comprised of the A 2 analog. Because of a diet of freshwater fish [12,22], these predators had a much wider variety of vitamin A esters in the liver than the most frequently analyzed laboratory species [15,16] or even marine seals [28] have. Törmä and Vahlquist [23] have identified small concentrations of alcoholic and esterified A 2 in mouse liver.…”

Section: Resultsmentioning

confidence: 99%

“…The major peaks of the A1 esters in the liver samples were identified by comparing their relative retention times to those of the synthesized standards in pure methanol and in the argentated mobile phase and by studying the mobility of the component peaks due to argentation. The A 2 esters can be distinguished from the A1 esters because of their different absorption spectra and maxima [22,23], which are 350 nm for the A 2 esters and 325 nm for the A 1 esters. Thus, detection using both wavelengths was needed.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of hepatic vitamins A₁, A₂, their fatty acyl esters, and vitamin E for biomonitoring mammals feeding on freshwater fish

Käkelä

Hyvärinen

2002

Enviro Toxic and Chemistry

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In tissues of freshwater fish-feeding mammals, 3,4-didehydroretinol (A2) is a major form of vitamin A. In mink liver, with organochlorine exposure, this analog has been found to decrease more than retinol (A1) and thus has potential as a sensitive freshwater biomarker. The presence of the analogs A1 and A2 as alcohol and different fatty acyl esters, which react to polychlorinated biphenyls differently, necessitates detailed analyses achieved by using direct extraction of tissue homogenate. In direct hexane extraction, compared to total levels of the vitamins obtained in the saponification procedure, a large proportion of the vitamins was released only after repeated and long-time vortex mixing with the extraction solvent. Thus, in tissue extraction, the use of internal standardization alone can lead to a rough underestimation of the levels of these fat-soluble vitamins. For analyses of vitamins A1 and A2 in liver, we applied the argentation high-performance liquid chromatography, which provided good separation of individual A1 and A2 fatty acyl esters. We report retention times for numerous esters of A1 and A2 and, to aid identification, the change in their retention properties after adding AgNO3 to the mobile phase. The argentation did not affect the recoveries of any forms of the retinoids studied but destroyed half the vitamin E. Despite selective acylation of fatty acids into the vitamin A esters, the fatty acids of the esters were the same as those found to be the major fatty acids in the gas-liquid chromatography of total lipids. The goal of this work was to create a methodology that is suitable for biomonitoring alcoholic and esterified vitamins A1 and A2 in tissues of freshwater fish-feeding mammals.

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High-performance liquid chromatography of the unsaponifiable from samples of marine and freshwater fish: fractionation and identification of retinol (vitamin A1) and dehydroretinol (vitamin A2) isomers

Cited by 34 publications

References 20 publications

HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues

HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues

Vitamins A₁, A₂, and E in minks exposed to polychlorinated biphenyls (Aroclor 1242®) and copper, VIA diet based on freshwater or marine fish

Analysis of hepatic vitamins A₁, A₂, their fatty acyl esters, and vitamin E for biomonitoring mammals feeding on freshwater fish

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High-performance liquid chromatography of the unsaponifiable from samples of marine and freshwater fish: fractionation and identification of retinol (vitamin A1) and dehydroretinol (vitamin A2) isomers

Cited by 34 publications

References 20 publications

HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues

HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues

Vitamins A1, A2, and E in minks exposed to polychlorinated biphenyls (Aroclor 1242®) and copper, VIA diet based on freshwater or marine fish

Analysis of hepatic vitamins A1, A2, their fatty acyl esters, and vitamin E for biomonitoring mammals feeding on freshwater fish

Contact Info

Product

Resources

About

Vitamins A₁, A₂, and E in minks exposed to polychlorinated biphenyls (Aroclor 1242®) and copper, VIA diet based on freshwater or marine fish

Analysis of hepatic vitamins A₁, A₂, their fatty acyl esters, and vitamin E for biomonitoring mammals feeding on freshwater fish