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Bioanalysis. 2015;7(9):1137-49. doi: 10.4155/bio.15.38.

Simultaneous determination of trimethoprim and sulfamethoxazole in dried plasma and urine spots.

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2Duke Clinical Research Institute, Duke University Medical Center, Durham, NC, USA.
3Department of Pediatrics, Riley Hospital for Children, Indiana University, Indianapolis, IN, USA.
5Department of Pediatrics, Medical University of South Carolina, Charleston, SC, USA.
6Kosair Charities Pediatric Clinical Research Unit, Department of Pediatrics, University of Louisville, Kosair Children's Hospital, Louisville, KY, USA.
7Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD, USA.
8Department of Neonatology, Wesley Medical Center, Wichita, KS, USA.
9OpAns, LLC, 4134 S Alston Ave # 101, Durham, NC 27713, USA.
10EMMES Corp., 401 N Washington St # 700, Rockville, MD 20850, USA.
11Department of Pediatrics, Duke University Medical Center, Durham, NC, USA.



Trimethoprim-sulfamethoxazole (TMP-SMX) is an antimicrobial drug combination commonly prescribed in children and adults. The study objectives were to validate and apply an HPLC-MS/MS method to quantify TMP-SMX in dried plasma spots (DPS) and dried urine spots (DUS), and perform a comparability analysis with liquid matrices.


For TMP the validated range was 100-50,000 ng/ml for DPS and 500-250,000 ng/ml for DUS; for SMX, the validated range was 1000-500,000 ng/ml for both DPS and DUS. Good agreement was noted between DPS/DUS and liquid plasma and urine samples for TMP, while only modest agreement was observed for SMX in both matrices.


A precise, accurate and reproducible method was developed to quantify TMP-SMX in DPS and DUS samples.

[Indexed for MEDLINE]
Free PMC Article

Conflict of interest statement

Financial & competing interest disclosure This work was funded under National Institute of Child Health and Human Development (NICHD) contract HHSN201000003I for the Pediatric Trials Network (PI: Benjamin) and HHSN27500006 (PI: Melloni, Cohen-Wolkowiez) for the Pharmacokinetics of Understudied Drugs Administered to Children per Standard of Care Study (POPS; protocol NICHD-2011-POP01). Research reported in this publication also was supported by the National Center for Advancing Translational Sciences of the National Institutes of Health (NIH) under award number UL1TR001117. The assay measuring TMP-SMX concentrations was performed at OpAns Laboratory (Durham, NC, USA). D Gonzalez is funded by training grant T32GM086330 from the National Institute of General Medical Sciences (NIGMS). D K Benjamin Jr receives support from the US government for his work in pediatric and neonatal clinical pharmacology (2K24HD058735–06, UL1TR001117, NICHD contract HHSN275201000003I and NIAID contract HHSN2722015000061); he also receives research support from Cempra Pharmaceuticals (subaward to HHSO100201300009C) for neonatal and pediatric drug development ( M Cohen-Wolkowiez receives support for research from the NIH (1R01-HD076676–01A1), the National Center for Advancing Translational Sciences of the NIH (UL1TR001117), the National Institute of Allergy and Infectious Disease (HHSN272201500006I and HHSN272201300017I), the National Institute for Child Health and Human Development of the NIH (HHSN275201000003I), the FDA (1U01FD004858–01), the Biomedical Advanced Research and Development Authority (HHSO100201300009C), the nonprofit organization Thrasher Research Fund ( and from industry (CardioDx and Durata Therapeutics) for drug development in adults and children ( The remaining authors have no funding to disclose. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. The authors thank A McMillan for her editorial assistance. A McMillan was funded by HHSN27500006 (PI: Melloni, Cohen-Wolkowiez).

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