Vitamin D analysis in plasma by high performance liquid chromatography (HPLC) with C30 reversed phase column and UV detection – Easy and acetonitrile-free

Hymøller, Lone; Jensen, Søren Krogh

doi:10.1016/j.chroma.2011.02.004

Cited by 53 publications

(45 citation statements)

References 18 publications

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“…The content of fat in the milk was determined by the gravimetric method, according to a modified Schmid-Bondzynski-Ratslaff procedure (NMKL, 1989). In the laboratory at the Department of Animal Science, Aarhus University (Tjele, Denmark), the content of 25OHD 3 in plasma was assessed in 1.5 mL of plasma by a method described elsewhere (Hymøller and Jensen, 2011).…”

Section: Short Communication: Artificial Ultraviolet B Light Exposurementioning

confidence: 99%

Short communication: Artificial ultraviolet B light exposure increases vitamin D levels in cow plasma and milk

Jakobsen

Jensen

Hymøller

et al. 2015

Journal of Dairy Science

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The number of dairy cows without access to pasture or sunlight is increasing; therefore, the content of vitamin D in dairy products is decreasing. Ultimately, declining vitamin D levels in dairy products will mean that dairy products are a negligible source of natural vitamin D for humans. We tested the ability of a specially designed UVB lamp to enhance the vitamin D3 content in milk from dairy cows housed indoors. This study included 16 cows divided into 4 groups. Each group was exposed daily to artificial UVB light simulating 1, 2, 3, or 4 h of summer sun at 56°N for 24 d, and the group with simulated exposure to 2 h of summer sun daily continued to be monitored for 73 d. We found a significant increase in 25-hydroxyvitamin D3 (25OHD3) levels in plasma as well as vitamin D3 and 25OHD3 levels in milk after daily exposure for 24 d in all treatment groups. Extending daily exposure to artificial UVB light to 73 d did not lead to an increase of vitamin D3 or 25OHD3 level in the milk. In conclusion, the change in production facilities for dairy cows providing cows with no access to pasture and sunlight causes a decrease of vitamin D levels in dairy products. This decrease may be prevented by exposing cows to artificial UVB light in the stable.

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Section: Short Communication: Artificial Ultraviolet B Light Exposurementioning

confidence: 99%

Short communication: Artificial ultraviolet B light exposure increases vitamin D levels in cow plasma and milk

Jakobsen

Jensen

Hymøller

et al. 2015

Journal of Dairy Science

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“…Working IS solutions were prepared under dimmed or yellow lighting and diluted from stock solutions in hexane (tocol, butylated or ethylated UN10 and UL10), EtOH (retinyl laurate), or THF (ECH) so that peak sizes were similar to the analytes in the plasma extracts. IS concentrations were checked spectrophotometrically using reported extinction coefficients (Table 1) [4,12,43,51] and were accepted if purity, as assessed by HPLC, was greater than 95%.…”

Section: Methodsmentioning

confidence: 99%

“…These reports, however, restrict their analyses to individual analytes [32–42] or a limited combination thereof [10,11,13–20,23,43–50] as a result of problems that arise when numerous LPM are simultaneously analyzed in a single chromatographic run. Examples of these problems include analyte co-elution and the extensive range of analyte concentrations, both of which make quantitation impossible because of interference or the inability to reach detectable levels.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous analysis of circulating 25-hydroxy-vitamin D3, 25-hydroxy-vitamin D2, retinol, tocopherols, carotenoids, and oxidized and reduced coenzyme Q10 by high performance liquid chromatography with photo diode-array detection using C18 and C30 columns alone or in combination

Franke

Morrison

Custer

et al. 2013

Journal of Chromatography A

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Circulating lipid-phase micronutrients (LPM) such as 25-hydroxylated D vitamers, retinol, tocopherols, carotenoids including their isomers, and coenzyme Q10 play important roles in health maintenance and disease prevention and can serve as useful biomarkers. We developed fast, affordable, and accurate HPLC assays that simultaneously measured all above LPM in a single run using UV/VIS detection at 265 nm, 295 nm, and 480 nm with (1) a C18 column alone; (2) a C30 column alone; or (3) each of these columns connected in series. The C18 column alone could separate all major LPM of interest in less than 17 min but insufficiently resolved the lycopene isomers, the 25-hydroxylated D vitamers, lutein from zeaxanthin and β- from γ-tocopherol. The C30 column alone separated all LPM of interest including many isomeric analytes but failed to resolve the Q10 compounds, which co-eluted with carotenoids. Connecting the C18 and C30 columns in series with a detector after the C30 column and a pressure resistant detector between the columns resulted in ideal resolution and accurate quantitation of all LPM of interest but required software capable of processing the acquired data from both detectors. Connecting the C18 and C30 columns in series with exclusively one detector after the C30 column resulted in carotenoid-Q10 interferences, however, this was remedied by heart-cutting 2D-LC with a 6-port valve between the columns, which resolved all analytes in 42 min. Faster run times led to some analytes not being resolved. Many variations of these methods are possible to meet the needs of individual requirements while minimizing sample material and turn-around-times.

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“…Based on well-established principle of alkaline hydro- unusually applied this approach for plasma/serum samples; in this procedure, alkalinisation was performed with sodium hydroxide and was found to be efficient for their purpose (74,82). Ethanolic KOH is preferred to aqueous KOH in the saponification of dairy products because it mixes well with fat and prevents formation of emulsions (32).…”

Section: Saponification (Sn)mentioning

confidence: 99%

“…Four analogues excluding epimers were separated using a RP column (57,82). Four (43,80) or five (36,75) analogues excluding epimers were separated using PTDA-derivatisation and a RP column.…”

Section: Separation and Detectionmentioning

confidence: 99%

Development of an assay for total vitamin D content in milk and application to study the effect of pasteurisation in breastmilk

Gomes¹

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Vitamin D analysis in plasma by high performance liquid chromatography (HPLC) with C30 reversed phase column and UV detection – Easy and acetonitrile-free

Cited by 53 publications

References 18 publications

Short communication: Artificial ultraviolet B light exposure increases vitamin D levels in cow plasma and milk

Short communication: Artificial ultraviolet B light exposure increases vitamin D levels in cow plasma and milk

Simultaneous analysis of circulating 25-hydroxy-vitamin D3, 25-hydroxy-vitamin D2, retinol, tocopherols, carotenoids, and oxidized and reduced coenzyme Q10 by high performance liquid chromatography with photo diode-array detection using C18 and C30 columns alone or in combination

Development of an assay for total vitamin D content in milk and application to study the effect of pasteurisation in breastmilk

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