MALDI-MS-Based Quantitative Analysis of Bioactive Forms of Vitamin D in Biological Samples

Ahn, Da-Hee; Kim, Hee-jin; Kim, Seong-Min; Jo, Sung-Hyun; Jeong, Jae-Hyun; Kim, Yun-Gon

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자료유형: 학술저널

저자정보: Ahn, Da-Hee (Department of Chemical Engineering, Soongsil University) Kim, Hee-jin (Department of Chemical Engineering, Soongsil University) Kim, Seong-Min (Department of Chemical Engineering, Soongsil University) Jo, Sung-Hyun (Department of Chemical Engineering, Soongsil University) Jeong, Jae-Hyun (Department of Chemical Engineering, Soongsil University) Kim, Yun-Gon (Department of Chemical Engineering, Soongsil University)

저널정보: 한국화학공학회 화학공학 화학공학 제58권 제1호

발행연도: 2020.1

수록면: 106 - 112 (7page)

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Analyzing vitamin D levels is important for monitoring health conditions because vitamin D deficiency is associated with various diseases such as rickets, osteomalacia, cardiovascular disorders and some cancers. However, vitamin D concentration in the blood is very low with optimal level of 75 nmol/L, making quantitative analysis difficult. The objective of this study was to develop a highly sensitive analysis method for vitamin D using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS). 25-hydroxyvitamin D (25(OH)D), which has been used as an indicator of vitamin D metabolites in human biofluids was chemically derivatized using a secosteroid signal enhancing tag (SecoSET) with powerful dienophile and permanent positive charge. The SecoSET-derivatized 25(OH)D provided good linearity ($R^2$ > 0.99) and sensitivity (limit of quantitation: 11.3 fmol). Chemical derivatization of deuterated 25-hydroxyvitamin $D_3$ ($d_6$-25(OH)D3) with SecoSET enabled absolute quantitative analysis using MALDI-MS. The highly sensitive method could be successfully applied into monitoring of quantitative changes of bioactive vitamin D metabolites after treatment with ketoconazole to inhibit $1{\alpha}$-hydroxylase reaction related to vitamin D metabolism in human breast cancer cells. Taken together, we developed a MALDI-MS-based platform that could quantitatively analyze vitamin D metabolites from cell products, blood and other biofluids. This platform may be applied to monitor various diseases associated with vitamin D deficiency such as rickets, osteomalacia and breast cancer.

#Vitamin D metabolites #25-hydroxyvitamin #MALDI-MS #Chemical derivatization #Quantitative analysis

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