Quantitative determination of vitamin D metabolites in plasma using UHPLC-MS/MS

Vitamin D is an important determinant of bone health at all ages. The plasma concentrations of 25-hydroxy vitamin D (25-OH D) and other metabolites are used as biomarkers for vitamin sufficiency and function. To allow for the simultaneous determination of five vitamin D metabolites, 25-OH D3, 25-OH D2, 24,25-(OH)2 D3, 1,25-(OH)2 D3, and 1,25-(OH)2 D2, in low volumes of human plasma, an assay using ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) was established. Plasma samples were spiked with isotope-labeled internal standards and pretreated using protein precipitation, solid-phase extraction (SPE) and a Diels–Alder derivatization step with 4-phenyl-1,2,4-triazoline-3,5-dione. The SPE recovery rates ranged from 55% to 85%, depending on the vitamin D metabolite; the total sample run time was <5 min. Mass spectrometry was conducted using positive ion electrospray ionization in the multiple reaction monitoring mode on a quadrupole–quadrupole-linear ion trap instrument after pre-column addition of methylamine to increase the ionization efficiency. The intra- and inter-day relative standard deviations were 1.6–4.1% and 3.7–6.8%, respectively. The limit of quantitation for these compounds was determined to be between 10 and 20 pg/mL. The 25-OH D results were compared with values obtained for reference materials (DEQAS). In addition, plasma samples were analyzed with two additional Diasorin antibody assays. All comparisons with conventional methods showed excellent correlations (r2 = 0.9738) for DEQAS samples, demonstrating the high degree of comparability of the new UHPLC-MS/MS technique to existing methods

Using multivariate analysis to predict the behaviour of soils under effluent irrigation

Onsite wastewater treatment systems aim to assimilate domestic effluent into the environment. Unfortunately failure of such systems is common and inadequate effluent treatment can have serious environmental implications. The capacity of a particular soil to treat wastewater will change over time. The physical properties influence the rate of effluent movement through the soil and its chemical properties dictate the ability to renovate effluent. A research project was undertaken to determine the role that physical and chemical soil properties play in predicting the long-term behaviour of soil under effluent irrigation and to determine if they have a potential function as early indicators of adverse effects of effluent irrigation on treatment sustainability. Principal Component Analysis (PCA) and Cluster Analysis grouped the soils independently of their soil classifications and allowed us to distinguish the most suitable soils for sustainable long term effluent irrigation and determine the most influential soil parameters to characterise them. Multivariate analysis allowed a clear distinction between soils based on the cation exchange capacities. This in turn correlated well with the soil mineralogy. Mixed mineralogy soils in particular sodium or magnesium dominant soils are the most susceptible to dispersion under effluent irrigation. The soil Exchangeable Sodium Percentage (ESP) was identified as a crucial parameter and was highly correlated with percentage clay, electrical conductivity, exchangeable sodium, exchangeable magnesium and low Ca:Mg ratios (less than 0.5)