High-performance liquid chromatographic separation of the fat-soluble vitamins in cod liver oil and feeds

A MicroPak-CN column with a methylene chloride—chloroforms-n-hexane (3:2:15) mobile phase separates the fat-soluble vitamins in cod liver oil and formulated feed. Powdered feed samples are dissolved in n-hexane—chloroform—ethanol (6:3.5:0.5) and centrifuged before injection. The oil is dissolved in the extraction solvent or mobile phase before injection. The naturally occurring vitamins are identified from their absorption spectra by stopped-flow spanning of the individual vitamins.     

Liquid-gel partitioning using Lipidex in the determination of polychlorinated biphenyls in cod liver oil

A technique was developed for transfer of fat and polychlorinated biphenyls from cod liver oil into the lipophilic gel Lipidex 5000. Subsequent elution of the gel separated about 60% of the fat from the sample. Following further purification on aluminium oxide and silica gel, toxic non-ortho- and mono-ortho-PCB congeners were isolated in two separate fractions on charcoal. Recoveries were studied by addition of twelve different PCB congeners to 0.2g of fat. The non-ortho-PCBs were labelled with ¹³C. The recoveries of 5–50 ng of the unlabelled compounds were 80–100% and those of 50–100 pg of the labelled compounds were 76–106%.     

Elucidation of triacylglycerols in cod liver oil by liquid chromatography electrospray tandem ion-trap mass spectrometry

Though liquid chromatography electrospray tandem mass spectrometry (LC-ESI-MS²) has been widely used in the structural elucidation of triacylglycerols (TAG) in vegetable oils, its potentiality for the identification of TAG molecules in omega-3 rich oils remains unexplored till date. Hence, this article investigates the applicability of LC-ESI-MS² for the structural characterization of naturally occurring TAG in cod liver oil without the TAG fractionation during the sample preparation. A computational algorithm was developed to automatically interpret the mass spectra and elucidate the TAG structures respectively. The results were compared against the lipase benchmark method. A principal component analysis study revealed that it is possible to discriminate genuine from adulterated cod liver oil.     

A new sample preparation technique for organochlorines in cod liver oil combining SPE and NP-HPLC with HRGC-ECD

An overview is given describing phenomena at heated microelectrodes where matter and heat energy are simultaneously emitted into the solution. With controlled electric heating, virtual “quiescent” periods as well as ones with constant streaming conditions are found that depend on the heating time. A close look at a permanently heated wire reveals a well defined structure with stationary concentration, temperature and flow rate profiles. The observed phenomena can be utilised for analytical measurements, e.g. with the novel method “Temperature Pulse Voltammetry” (TPV).     

A new sample preparation technique for organochlorines in cod liver oil combining SPE and NP-HPLC with HRGC-ECD

The analysis of semivolatile organochlorines (polychlorinated biphenyls and chlorinated pesticides) in less polluted biomaterials requires specific strategies in controlling the blank in sample preparation. The procedure described here allows to decrease significantly the level of contamination during the clean-up step of fish oil. Solid-phase-extraction (SPE) on LiChrolut EN and normal phase HPLC in the normal- and the backflush-mode were used to reduce the amount of solvents needed and the analysis time compared to established clean-up procedures. With a certified reference material (BCR-CRM 349; Cod Liver Oil) the precision and effectiveness of the new method were validated. Recovery rates of the Internal Standards (PCB 103 and TCN) lay between 75% and 90% at the microg/kg lipid level. The quantitative analyses were carried out by high resolution gas chromatography with electron capture detector (HRGC-ECD).     

High-performance liquid chromatography of fat-soluble vitamins. Simultaneous quantitative analysis of vitamins D2, D3 and E. Study of percentage recoveries of vitamins from cod liver oil

Vitamins D2, D3 and E were resolved and quantified by applying reversed-phase high-performance liquid chromatography to extracts of cod liver oil. The method, using two reversed-phase C18 columns and a ternary mixture of acetonitrile, methanol and water as the eluent resolved all fat-soluble vitamins well, including the pair D2-D3. The extraction procedure was studied; the recoveries, using two different solvents (hexane and diethyl ether) for extractions were 60.6 +/- 1.0 and 77.1 +/- 1.1, 56.9 +/- 1.2 and 74.8 +/- 0.8, and 14.1 +/- 0.7 and 89.8 +/- 1.4% for vitamins D2, D3 and E, respectively.     

Synthesis of vitamin D(3) and calcitriol dimers as potential chemical inducers of vitamin D receptor dimerization

The design and synthesis of vitamin D(3) dimers 2 and 3 and 1 alpha, 25-dihydroxyvitamin D(3) (calcitriol) dimers 4 and 5 are described. The dimers were designed with a view to doubly binding the vitamin D receptor (VDR) and inducing the receptor homodimerization. In the dimers the units are linked through the C-11 position in ring C by an alkyl side chain of six or 10 carbon atoms, far from the hydroxy groups responsible for the VDR binding. The linker is formed by olefin metathesis of an olefinic side chain at the C-11 position introduced by stereoselective cuprate addition. The synthesis, which is both short and convergent, uses the Wittig-Horner approach to construct the vitamin D triene system and allows the preparation of dimers with a linker of modulated length with the purpose of optimizing the vitamin D(3)-VDR interaction.     

Quantitative determination of toxic mono- and non-ortho polychlorinated biphenyls in cod liver oil after selective liquid chromatographic separation

Summary By application of chromatographic column filled with Supelclean ENVI-Carb/Celite and elution with three solvents of different polarity three PCB fractions were obtained. Fraction A contained poly-ortho PCBs, Fraction B mono-ortho PCBs, and Fraction C non-ortho PCBs. The Supelclean ENVI-Carb/Celite column was used in combination with a sample preparation procedure for pre-cleaning of acid-stable chlorinated hydrocarbons such as DDT and its metabolites, HCH isomers, and regulation-relevant PCB congeners. The method was optimized using standard solutions of 55 PCB congeners, 8 chlorinated pesticides and contaminated cod liver oil samples. The influence of traces of remaining matrix on the elution profile of the organochlorine compounds on Supelclean ENVI-Carb/Celite was observed. Quantitation was carried out by GC-ECD with fused silica capillary columns of different polarity.     

Determination of vitamin D3 in cod-liver oil by high-performance liquid chromatography

The present British Pharmacopoeia monograph for cod-liver oil requires a bioassay for the vitamin D3 content which is both time-consuming and complex. Alternative assays employing chromatographic procedures have been described but all these involve prior saponification of the oil. A selective extraction for vitamin D3 without the need for saponification is reported in this paper. The extraction utilizes only chromatographic assay using argentation on reversed-phase silica, with vitamin D2 as the internal standard. Reproducibility of injection gave a coefficient of variation of 0.6%, and repeatability of extraction for six samples gave a coefficient of variation of 6.8%.     

Sun Exposure and Vitamin D Status as Northeast Asian Migrants Become Acculturated to Life in Australia

Vitamin D deficiency is more common in Northeast‐Asian immigrants to western countries than in the local population; prevalence equalizes as immigrants adopt the host country's culture. In a community‐based study of 100 Northeast‐Asian immigrants in Canberra, Australia, we examined predictors of vitamin D status, its association with indicators of acculturation (English language use; time since migration) and mediators of that association. Participants completed a sun and physical activity diary and wore an electronic ultraviolet radiation (UVR) dosimeter for 7 days. Skin colour was measured by reflectance spectrophotometry. Serum concentrations of 25‐hydroxyvitamin D (25(OH)D) and cardio‐metabolic biomarkers were measured on fasting blood. In a multiple linear regression model, predictors for 25(OH)D concentration were season of blood collection, vitamin D supplementation, UVR exposure, body mass index, physical activity and having private health insurance (R² = 0.57). Greater acculturation was associated with lower risk of vitamin D deficiency (de‐seasonalized 25(OH)D level <50 nmol L^?1) (Adjusted Odds Ratio (AOR): 0.22 [95%CI 0.04–0.96]); this association was statistically mediated by physical activity and time outdoors. Vitamin D deficiency was associated with higher total cholesterol levels (>5.0 mmol L^?1) (AOR: 7.48 [95%CI 1.51–37.0]). Targeted public health approaches are required to manage the high prevalence of vitamin D deficiency in migrants retaining a traditional lifestyle.     

Interaction between vitamin D receptor and vitamin D ligands: two-dimensional alanine scanning mutational analysis

We present a new method to investigate the details of interaction between vitamin D nuclear receptor (VDR) and various ligands, namely a two-dimensional alanine scanning mutational analysis. In this method, the transactivation of various ligands is studied in conjunction with a series of alanine scanning mutations of the residues lining the ligand binding pocket (LBP) of VDR, and the complete set of results is profiled in a patch table. We investigated examples from four structurally diverse groups of known VDR ligands: the native vitamin D hormone and two compounds with the same side chain configuration; four 20-epi compounds; three 19-nor compounds; and two nonsecosteroids. The patch table of the results indicates characteristics of each group in terms of its interaction with 18 LBP residues. We demonstrate the validity of this approach by application to docking studies of the two nonsecosteroids.     

The Sun and Jupiter as a binary system

I view the solar system in my paper as basically a double star, with the sun and Jupiter as components. Although the acceptance of this model may still be a philosophical choice, I introduce in my paper powerful arguments in favor of its support. Those arguments cover a wide range of conceptual, physical, and mathematical analysis.     

Enhancement of pullulan production by aureobasidium pullulans in batch culture using olive oil and sucrose as carbon sources

The production of pigment-free pullulan byAureobasidium pullulans, using olive oil and sucrose as carbon (C) sources, in shake flasks, was investigated. Optimum medium composition for pullulan elaboration was 80 g/L sucrose, 25 mL/L olive oil, 5 mL/L Tween-80, 10 g/L glutamic acid, and an initial pH of 5.5. Maximum pullulan concentration (51.5 g/L), productivity (8.6 g/L·d), and yield (80.3%) were achieved under these conditions after 120 h of fermentation. The principal advantage of using olive oil and sucrose simultaneously as C sources was the elimination of the inhibitory effect of high sucrose concentrations (> 60 g/L) on pullulan production by the microorganism. Structural characterization by¹³C-NMR, monosaccharide, and methylation analyses, and pullulanase digestion, combined with size-exclusion chromatography, confirmed the identity of pullulan and the homogeneity of the released polysaccharide in the fermentation broths. There were no significant differences in structure between pullulan samples isolated from either olive oil-supplemented media or olive oil-free media. The molecular size of pullulan from the combined olive oil-sucrose fermentation was slightly lower (1.1 X 10⁶) than that of conventional fermentation with sucrose as a single C source (1.4 X 10⁶). Lowering the initial pH of the medium resulted in increased molecular size for the released polymer, but a lower pullulan yield.     

Quantification of physiological levels of vitamin D3 and 25‐hydroxyvitamin D3 in porcine fat and liver in subgram sample sizes

Most methods for the quantification of physiological levels of vitamin D₃ and 25‐hydroxyvitamin D₃ are developed for food analysis where the sample size is not usually a critical parameter. In contrast, in life science studies sample sizes are often limited. A very sensitive liquid chromatography with tandem mass spectrometry method was developed to quantify vitamin D₃ and 25‐hydroxyvitamin D₃ simultaneously in porcine tissues. A sample of 0.2–1 g was saponified followed by liquid–liquid extraction and normal‐phase solid‐phase extraction. The analytes were derivatized with 4‐phenyl‐1,2,4‐triazoline‐3,5‐dione to improve the ionization efficiency by electrospray ionization. The method was validated in porcine liver and adipose tissue, and the accuracy was determined to be 72–97% for vitamin D₃ and 91–124% for 25‐hydroxyvitamin D₃. The limit of quantification was <0.1 ng/g, and the precision varied between 1.4 and 16% depending on the level of spiking. The small sample size required for the described method enables quantification of vitamin D₃ and 25‐hydroxyvitamin D₃ in tissues from studies where sample sizes are limited.