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Can dried blood spots be used to accurately measure vitamin D metabolites?
•Vitamin D metabolites were accurately quantified from DBS samples using LC-MS/MS.•The assay discriminated the C-3 epimer from standard vitamin D metabolites.•Haematocrit remains a challenge when using DBS to quantify plasma metabolites. Where conventional blood sampling is challenging, dried blood...
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| Published in: | Clinica chimica acta 2021-07, Vol.518, p.70-77 |
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| Main Authors: | , , , , , , |
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| container_end_page | 77 |
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| container_title | Clinica chimica acta |
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| creator | Binks, Michael J. Bleakley, Amy S. Rathnayake, Geetha Pizzutto, Susan Chang, Anne B. McWhinney, Brett Ungerer, Jacobus |
| description | •Vitamin D metabolites were accurately quantified from DBS samples using LC-MS/MS.•The assay discriminated the C-3 epimer from standard vitamin D metabolites.•Haematocrit remains a challenge when using DBS to quantify plasma metabolites.
Where conventional blood sampling is challenging, dried blood spots (DBS) provide a practical sample alternative for measuring vitamin D levels. Our study aimed to develop and evaluate a clinical pathology service-based assay suitable for measuring vitamin D in batches of DBS samples collected remote to the testing site.
A high throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with derivatisation was developed to measure 25-hydroxyvitamin D metabolites (25OHD3, 25OHD2 and 3-epi-25OHD3) in DBS samples. The assay was validated using paired DBS and plasma samples from 37 healthy adults.
The assay reproducibly ( |
| doi_str_mv | 10.1016/j.cca.2021.03.003 |
| format | article |
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Where conventional blood sampling is challenging, dried blood spots (DBS) provide a practical sample alternative for measuring vitamin D levels. Our study aimed to develop and evaluate a clinical pathology service-based assay suitable for measuring vitamin D in batches of DBS samples collected remote to the testing site.
A high throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with derivatisation was developed to measure 25-hydroxyvitamin D metabolites (25OHD3, 25OHD2 and 3-epi-25OHD3) in DBS samples. The assay was validated using paired DBS and plasma samples from 37 healthy adults.
The assay reproducibly (<11.5% coefficient of variation) quantified 25OHD3 (range 1–300 nmol/L), 25OHD2 (range 2–300 nmol/L) and 3-epi-25OHD3 (range 1–200 nmol/L) in DBS samples. The 25OHD3 metabolite was detected in all DBS samples, 3-epi-25OHD3 in six plasma (range 2.1–6.3 nmol/L) and paired DBS samples, and 25OHD2 was not detected. Concentrations of 25OHD3 were highly correlated between paired samples: capillary DBS and venous plasma (r = 0.92), venous DBS and venous plasma (r = 0.93), and capillary DBS and venous DBS (r = 0.97). Ordinary least squares regression was used to characterise (β = 0.81) and correct the systematic bias in DBS data (compared to paired plasma). Thereafter, Bland-Altman analysis demonstrated robust agreement between sample-methods.
This simple and rapid DBS-based LC-MS/MS assay accurately quantified serum vitamin D metabolites using a paired-sample ‘bridging strategy’ to correct for the inherent sample-method bias.</description><identifier>ISSN: 0009-8981</identifier><identifier>EISSN: 1873-3492</identifier><identifier>DOI: 10.1016/j.cca.2021.03.003</identifier><identifier>PMID: 33713691</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>25-hydroxyvitamin D ; 3-epi-25-hydroxyvitamin D ; Dried blood spot ; LC/MS/MS ; Vitamin D</subject><ispartof>Clinica chimica acta, 2021-07, Vol.518, p.70-77</ispartof><rights>2021 Elsevier B.V.</rights><rights>Copyright © 2021 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c353t-d8a40948c0a5632779f8170caecdfd9e456c4fd6e353009747aeb03c31692c2e3</citedby><cites>FETCH-LOGICAL-c353t-d8a40948c0a5632779f8170caecdfd9e456c4fd6e353009747aeb03c31692c2e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33713691$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Binks, Michael J.</creatorcontrib><creatorcontrib>Bleakley, Amy S.</creatorcontrib><creatorcontrib>Rathnayake, Geetha</creatorcontrib><creatorcontrib>Pizzutto, Susan</creatorcontrib><creatorcontrib>Chang, Anne B.</creatorcontrib><creatorcontrib>McWhinney, Brett</creatorcontrib><creatorcontrib>Ungerer, Jacobus</creatorcontrib><title>Can dried blood spots be used to accurately measure vitamin D metabolites?</title><title>Clinica chimica acta</title><addtitle>Clin Chim Acta</addtitle><description>•Vitamin D metabolites were accurately quantified from DBS samples using LC-MS/MS.•The assay discriminated the C-3 epimer from standard vitamin D metabolites.•Haematocrit remains a challenge when using DBS to quantify plasma metabolites.
Where conventional blood sampling is challenging, dried blood spots (DBS) provide a practical sample alternative for measuring vitamin D levels. Our study aimed to develop and evaluate a clinical pathology service-based assay suitable for measuring vitamin D in batches of DBS samples collected remote to the testing site.
A high throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with derivatisation was developed to measure 25-hydroxyvitamin D metabolites (25OHD3, 25OHD2 and 3-epi-25OHD3) in DBS samples. The assay was validated using paired DBS and plasma samples from 37 healthy adults.
The assay reproducibly (<11.5% coefficient of variation) quantified 25OHD3 (range 1–300 nmol/L), 25OHD2 (range 2–300 nmol/L) and 3-epi-25OHD3 (range 1–200 nmol/L) in DBS samples. The 25OHD3 metabolite was detected in all DBS samples, 3-epi-25OHD3 in six plasma (range 2.1–6.3 nmol/L) and paired DBS samples, and 25OHD2 was not detected. Concentrations of 25OHD3 were highly correlated between paired samples: capillary DBS and venous plasma (r = 0.92), venous DBS and venous plasma (r = 0.93), and capillary DBS and venous DBS (r = 0.97). Ordinary least squares regression was used to characterise (β = 0.81) and correct the systematic bias in DBS data (compared to paired plasma). Thereafter, Bland-Altman analysis demonstrated robust agreement between sample-methods.
This simple and rapid DBS-based LC-MS/MS assay accurately quantified serum vitamin D metabolites using a paired-sample ‘bridging strategy’ to correct for the inherent sample-method bias.</description><subject>25-hydroxyvitamin D</subject><subject>3-epi-25-hydroxyvitamin D</subject><subject>Dried blood spot</subject><subject>LC/MS/MS</subject><subject>Vitamin D</subject><issn>0009-8981</issn><issn>1873-3492</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><recordid>eNp9kE1LxDAQhoMoun78AC-So5fWSdKmLR5E1m8EL3oOaTKFLO1mTVLBf29k1aOnYYZnXmYeQk4ZlAyYvFiVxuiSA2cliBJA7JAFaxtRiKrju2QBAF3Rdi07IIcxrnJbgWT75ECIhgnZsQV5Wuo1tcGhpf3ovaVx41OkPdI55lnyVBszB51w_KQT6jgHpB8u6cmt6U2eJN370SWMV8dkb9BjxJOfekTe7m5flw_F88v94_L6uTCiFqmwra6gq1oDupaCN003tKwBo9HYwXZY1dJUg5WY6Xx_UzUaexBGMNlxw1EckfNt7ib49xljUpOLBsdRr9HPUfEaGJc1FzKjbIua4GMMOKhNcJMOn4qB-laoViorVN8KFQiVFeads5_4uZ_Q_m38OsvA5RbA_OSHw6Cicbg2aF1Ak5T17p_4LwrDgCU</recordid><startdate>20210701</startdate><enddate>20210701</enddate><creator>Binks, Michael J.</creator><creator>Bleakley, Amy S.</creator><creator>Rathnayake, Geetha</creator><creator>Pizzutto, Susan</creator><creator>Chang, Anne B.</creator><creator>McWhinney, Brett</creator><creator>Ungerer, Jacobus</creator><general>Elsevier B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20210701</creationdate><title>Can dried blood spots be used to accurately measure vitamin D metabolites?</title><author>Binks, Michael J. ; Bleakley, Amy S. ; Rathnayake, Geetha ; Pizzutto, Susan ; Chang, Anne B. ; McWhinney, Brett ; Ungerer, Jacobus</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c353t-d8a40948c0a5632779f8170caecdfd9e456c4fd6e353009747aeb03c31692c2e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>25-hydroxyvitamin D</topic><topic>3-epi-25-hydroxyvitamin D</topic><topic>Dried blood spot</topic><topic>LC/MS/MS</topic><topic>Vitamin D</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Binks, Michael J.</creatorcontrib><creatorcontrib>Bleakley, Amy S.</creatorcontrib><creatorcontrib>Rathnayake, Geetha</creatorcontrib><creatorcontrib>Pizzutto, Susan</creatorcontrib><creatorcontrib>Chang, Anne B.</creatorcontrib><creatorcontrib>McWhinney, Brett</creatorcontrib><creatorcontrib>Ungerer, Jacobus</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Clinica chimica acta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Binks, Michael J.</au><au>Bleakley, Amy S.</au><au>Rathnayake, Geetha</au><au>Pizzutto, Susan</au><au>Chang, Anne B.</au><au>McWhinney, Brett</au><au>Ungerer, Jacobus</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Can dried blood spots be used to accurately measure vitamin D metabolites?</atitle><jtitle>Clinica chimica acta</jtitle><addtitle>Clin Chim Acta</addtitle><date>2021-07-01</date><risdate>2021</risdate><volume>518</volume><spage>70</spage><epage>77</epage><pages>70-77</pages><issn>0009-8981</issn><eissn>1873-3492</eissn><abstract>•Vitamin D metabolites were accurately quantified from DBS samples using LC-MS/MS.•The assay discriminated the C-3 epimer from standard vitamin D metabolites.•Haematocrit remains a challenge when using DBS to quantify plasma metabolites.
Where conventional blood sampling is challenging, dried blood spots (DBS) provide a practical sample alternative for measuring vitamin D levels. Our study aimed to develop and evaluate a clinical pathology service-based assay suitable for measuring vitamin D in batches of DBS samples collected remote to the testing site.
A high throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with derivatisation was developed to measure 25-hydroxyvitamin D metabolites (25OHD3, 25OHD2 and 3-epi-25OHD3) in DBS samples. The assay was validated using paired DBS and plasma samples from 37 healthy adults.
The assay reproducibly (<11.5% coefficient of variation) quantified 25OHD3 (range 1–300 nmol/L), 25OHD2 (range 2–300 nmol/L) and 3-epi-25OHD3 (range 1–200 nmol/L) in DBS samples. The 25OHD3 metabolite was detected in all DBS samples, 3-epi-25OHD3 in six plasma (range 2.1–6.3 nmol/L) and paired DBS samples, and 25OHD2 was not detected. Concentrations of 25OHD3 were highly correlated between paired samples: capillary DBS and venous plasma (r = 0.92), venous DBS and venous plasma (r = 0.93), and capillary DBS and venous DBS (r = 0.97). Ordinary least squares regression was used to characterise (β = 0.81) and correct the systematic bias in DBS data (compared to paired plasma). Thereafter, Bland-Altman analysis demonstrated robust agreement between sample-methods.
This simple and rapid DBS-based LC-MS/MS assay accurately quantified serum vitamin D metabolites using a paired-sample ‘bridging strategy’ to correct for the inherent sample-method bias.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>33713691</pmid><doi>10.1016/j.cca.2021.03.003</doi><tpages>8</tpages></addata></record> |
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| source | ScienceDirect Journals |
| subjects | 25-hydroxyvitamin D 3-epi-25-hydroxyvitamin D Dried blood spot LC/MS/MS Vitamin D |
| title | Can dried blood spots be used to accurately measure vitamin D metabolites? |
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