Serum biomarkers of the calcium-deficient rats identified by metabolomics based on UPLC/Q-TOF MS/MS
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RESEARCH
Serum biomarkers of the calcium‑deficient rats identified by metabolomics based on UPLC/ Q‑TOF MS/MS Fanyu Meng1†, Lina Fan2†, Lin Sun3, Qingli Yu1, Maoqing Wang1* and Changhao Sun1*
Abstract Background: We previously identified the urinary biomarkers to diagnose calcium deficiency and nutritional rickets by ultra-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (UPLC/Q-TOF MS/MS). To find biomarkers of calcium deficiency and further confirm these biomarkers in serum, we performed serum metabolomics analysis of calcium-deficient rats. Methods: A calcium-deficient rat model was established with a low-calcium diet for 12 weeks. Serum metabolomics based UPLC/Q-TOF MS/MS and multivariate statistical analysis was performed to identify the alterations in metabolites associated with calcium deficiency in rats. Results: Bone mineral density, serum parathyroid hormone and alkaline phosphatase were significantly decreased in the low-calcium diet group (LCG) compared to the normal calcium diet group (NCG). Serum metabolic-profiling analysis could definitively distinguish between the LCG and NCG and identified 24 calcium-deficient biomarkers. Three metabolites (indoxyl sulfate, phosphate, and taurine) of the 24 biomarkers were found in our previous urinary metabolomics study of rats with a calcium deficiency and nutritional rickets. The areas under the curve (AUCs) of these three biomarkers were greater than 0.8, and the combination of any two biomarkers was higher than 0.95. Conclusion: Dietary calcium deficiency induced the alterations of metabolites in the serum of rats, and the three identified biomarkers had relatively high diagnostic values for calcium deficiency in rats. Keywords: Biomarkers, Calcium deficiency, Metabolomics, UPLC/Q-TOF MS/MS, Serum Introduction Calcium is a major constituent of bones and teeth and also plays an essential role as a second messenger in cellsignaling pathways [1]. An adequate intake of calcium is indispensable for human health. Calcium deficiency is a worldwide nutritional-deficiency public-health problem [2]. Similar to other countries, calcium deficiency *Correspondence: [email protected]; Sun2002changhao@yahoo. com † Fanyu Meng and Lina Fan have contributed equally to his work. 1 National Key Disciplines of Nutrition and Food Hygiene, Department of Nutrition and Food Hygiene, School of Public Health, Harbin Medical University, Harbin, China Full list of author information is available at the end of the article
is common in China [3, 4]. Over the past 20 years, the calcium intake of Chinese adults has not been sufficient, with a large gap between the actual intake and recommended intake of calcium. The proportions of citizens who satisfy the recommended appropriate intake (RNI) have been reported to be less than 7% in adult males, less than 5% in adult females and less than 4% in older adults over the age of 50 [5, 6]. Long-term calcium deficiency affects the health of the body and decreases the bone mineral density (
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