![]() ![]() Low 25-OH vitamin D concentrations are also associated with decreased bone mineral density. Elevated parathyroid (PTH) levels, especially in elderly vitamin D-deficient adults, can result in osteomalacia, increased bone turnover, reduced bone mass, and risk of bone fractures. Also, vitamin D deficiency is a common cause of secondary hyperparathyroidism. This condition led physicians to assess vitamin D concentrations in their patients (Holick 2007 Wang 2009). In recent years, clinical findings showed that low vitamin D levels are associated with various non-skeletal diseases. 2011) and cardio-vascular (McGreevy and Williams 2011) and autoimmune diseases (Ascherio et al. Vitamin D deficiency has been implicated in various cancers (Buttigliero et al. Vitamin D is important for the function of the cardiovascular, immune, and reproductive systems (Norman 2008 Dusso et al. Vitamin D, a fat-soluble steroid hormone produced in the skin by exposure to sunlight or supplied via dietary sources, plays a vital role in skeletal metabolism and calcium homeostasis throughout life (Prentice et al. The simple sample preparation and ease of use make it practical for the routine clinical laboratory. The HPLC assay demonstrates excellent linearity, acceptable accuracy and precision, and good agreement with a validated ECL method. Comparisons of results demonstrated good agreement between HPLC and ECL methods ( R 2 = 0.883). The mean was 37.875 ng/ml, the standard deviation was 0.22, and the coefficient of variation was 0.58%. Intra-assay precision was determined by extracting and quantifying 10 serum replicates from one patient. The method was linear for the lower limit of quantification from 3 ng/l up to at least 200 ng/l for 25(OH)D3, with the following equation for the regression line: y = 0.172 X + 2.45 ( R 2 = 0.989). Our HPLC results were compared with a validated electrochemiluminescence method. To assess its analytical performance, we used the RECIPE HPLC Complete Kit and an HPLC-UV instrument. It is also supplied in the diet and plays a pivotal role in calcium homeostasis and skeletal metabolism throughout life. Vitamin D is a fat-soluble steroid hormone precursor that is mainly produced in the skin by exposure to sunlight. The aim of the present study was to develop a robust and easy to use high performance liquid chromatography (HPLC) to analyze 25(OH)D3 in human serum. ![]()
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