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Ingredients

Analytical Accuracy of Fat-Soluble

Vitamins with Agilent Technologies

ou need the Agilent
fortitude for accurate and
Ysimultaneous quantitation of
fat-soluble vitamins, ergocalciferol
and cholecalciferol in Edible of the different dilutions used for

Oil. Vitamin D deficiency is one the calibration curves are merged.
of the most common nutrient System reproducibility was checked

deficiencies and is a well-known using six repeated injections of
cause of rickets, a bone disease An analytical method was developed extracted oil sample spiked at 50
seen in children. Taking vitamin using the Agilent 6470 triple quadru- ng/g level. The response %RSDs of
D supplements has been shown to pole LC/MS system with APCI ioni- six injections of vitamin D2 and
have numerous benefits related to zation installed, to ensure accuracy vitamin D3 were found to be 4.7%

cancer, bone health, mental health and simultaneous quantitation of and 3.9% respectively. To evaluate
and autoimmune diseases. Vitamin these vitamins. Sample preparation the effi ciency of the sample
D from the diet or dermal synthesis is based on saponifi cation, followed extraction, spike recovery studies
from sunlight is biologically inactive by extraction with hexane. The evap- were conducted for both vitamin D2
and its activation requires enzymatic orated hexane residue was reconsti- and vitamin D3 in oil samples at two
conversion (hydroxylation) in the tuted in methanol before instrumen- different spike levels: 50 and 100 ng/g,
liver and kidney. Minor differences tal analysis. The sample preparation Average recoveries of 91.5 % and 87 %
in the chemistry of side chains involved a saponifi cation process, for vitamin D2 and vitamin D3 were
between the two forms of vitamin which was the addition of concen- calculated.
D results in differences in the site trated potassium hydroxide to an
of hydroxylation and leads to the ethanol solution of sample, followed A sensitive LC/MS/MS method was
production of unique biologically by reflux heating at 70°C. This pro- developed for the quantifi cation of

active metabolites. cess breaks down the fat to release two forms of vitamin D: ergocalciferol
the matrix-bound vitamins. These (vitamin D2) and cholecalciferol
Ergocalciferol and cholecalciferol released vitamins were extracted (vitamin D3), fat soluble vitamins.
are two different forms of vitamin with hexane. The extracted hexane This multiple reaction monitoring
D and it is one of the essential fat- layer was evaporated completely and (MRM) based method was found to

soluble vitamins that play a major reconstituted in methanol. be specific in a complex matrix such
role in many biological activities as edible oil.
in the human body. A major source A LC/MS method was developed on an
of vitamin D is sunlight exposure Agilent 6470 LC/MS triple quadrupole Adopted APCI ionization technology
to the skin. Most people get their system installed with an APCI source in positive ionization mode reduced
vitamin D requirements through a operated in positive ionization mode. the matrix effect. Sample preparation

combination of solar conversion and Matrix-matched calibration curves was based on saponification of the
by maintaining a balanced diet. Liver, were made for both ergocalciferol oil sample followed by liquid-liquid

fish, eggs and milk together with and cholecalciferol from 10 to 500 extraction with hexane. The method
few other dairy products are rich in ng/g. Calibration curves were found demonstrated good reproducibility
vitamin D. In dairy products, vitamin to be linear within the previously and decent recovery. This method
D exists as cholecalciferol. To mentioned concentration ranges, can be adopted by commercial
maintain the vitamin D requirement with a minimum correlation testing labs involved in the analysis
of the population, ergocalciferol and coeffi cient of 0.9940. Different of vitamin D, required for nutritional
cholecalciferol are used to fortify concentration levels are prepared labeling of edible oils.
food ingredients in cooking oil as for both vitamin D2 and vitamin
it reduce issues related to vitamin D3 and plotted to generate the To know more download the App
defi ciency. calibration curves. Chromatograms Note.

Food Marketing & Technology 26 February 2020