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The dried blood spot (DBS) sampling technique is an attractive sample collection technique for infants. The present study reports the development and validation of a highly sensitive and precise method for quantification of thiamine diphosphate from DBS. The method utilizes full\u2010spot analysis of a volumetrically deposited 40 \u03bcl DBS. The analyte was extracted from the DBS using 50% methanol and then derivatized using potassium ferricyanide to thiochrome. Separation was achieved with the help of an Inertsil ODS C<jats:sub>18<\/jats:sub> column (5.0\u2009\u03bcm, 250\u2009\u00d7\u20094.6\u2009mm) using 150\u2009m<jats:sc>m<\/jats:sc> phosphate buffer pH\u20097\u2013acetonitrile (90:10, % v\/v) as the mobile phase. The use of a fluorimetric detector gave a good response to the thiochrome derivative offering good sensitivity for the method. The excitation and emission wavelengths were 367\u2009and 435\u2009nm, respectively. The limit of detection and lower limit of quantification were 5 and 10\u2009ng\/ml, respectively. Linearity was demonstrated from 10 to 1000\u2009ng\/ml, and precision (CV) was &lt;12.08%, at all tested quality control levels. The method accuracy was 89.34\u2013118.89% with recoveries &gt;80%. Bland\u2013Altman analysis of DBS sampling vs. whole blood demonstrated a mean bias of only 1.16\u2009ng\/ml, with a majority of the 60 investigated patient samples lying within 7.2% of the corresponding concentration measured in blood, thereby meeting the clinical desirable biological specification criterion and showing that the two methods are comparable.<\/jats:p>","DOI":"10.1002\/bmc.4668","type":"journal-article","created":{"date-parts":[[2019,7,28]],"date-time":"2019-07-28T23:22:37Z","timestamp":1564356157000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Development and validation of a dried blood spot test for thiamine deficiency among infants by HPLC\u2013fluorimetry"],"prefix":"10.1002","volume":"33","author":[{"given":"Elizabeth Mary","family":"Mathew","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Quality Assurance Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education  Manipal India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pradnya","family":"Sakore","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Quality Assurance Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education  Manipal India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3467-6821","authenticated-orcid":false,"given":"Leslie","family":"Lewis","sequence":"additional","affiliation":[{"name":"Department of Paediatrics Kasturba Medical College, Manipal Academy of Higher Education  Manipal India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kalaivani","family":"Manokaran","sequence":"additional","affiliation":[{"name":"Department of Medical Laboratory Technology Manipal Academy of Higher Education  Manipal India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1200-889X","authenticated-orcid":false,"given":"Pragna","family":"Rao","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Kasturba Medical College, Manipal Academy of Higher Education  Manipal India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5341-0981","authenticated-orcid":false,"given":"Sudheer","family":"Moorkoth","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Quality Assurance Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education  Manipal India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2019,9,4]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/cc10495"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0065-2423(08)60075-X"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003581"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/ajcn\/74.6.712"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-384947-2.00690-5"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpeds.2012.05.006"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1515\/cclm-2017-0054"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcrc.2010.03.003"},{"key":"e_1_2_8_10_1","volume-title":"Bioanalytical method validation guidance for industry","author":"Food and Drug administration","year":"2018"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2014.11.001"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1177\/0884533614561790"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.4155\/bio.14.185"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-4975(99)00144-7"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2017.06.048"},{"key":"e_1_2_8_16_1","first-page":"695","volume-title":"Clinical biochemistry of domestic animals","author":"Kaneko J. 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