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Quantification of Serum Sulfamethoxazole and Trimethoprim by Ultra-Fast SPE-MS/MS.
Ther Drug Monit. 2020 Jun 29 [Online ahead of print]TD

Abstract

BACKGROUND

The combination of trimethoprim (TMP) and sulfamethoxazole (SMX) is used to treat a number of bacterial infections. TMP/SMX concentrations in serum are conventionally monitored using high-performance liquid chromatography (HPLC) or liquid chromatography tandem mass spectrometry (LC-MS/MS). These methods require laborious manual extraction techniques and relatively long sample analysis times, necessitating the development of a simple, high-throughput method. A simple, high-throughput method to measure TMP/SMX using ultra-fast solid-phase extraction-tandem mass spectrometry (SPE-MS/MS) has been developed.

METHODS

Calibration standards, quality control materials, and patient samples were precipitated with acetonitrile containing isotopically labeled internal standards. Samples were vortexed, centrifuged for 5 minutes at 2053×g, and the resulting supernatant diluted in aqueous mobile phase and injected onto the C18 SPE cartridge. MS/MS analysis was carried out by electrospray ionization in positive ion mode at a rate of <20 seconds per sample. A five-point linear 1/x calibration curve was used to calculate sample concentrations.

RESULTS

The intra-assay precision coefficients of variation (PCVs) were <6% and <7% for SMX and TMP, respectively, and <10% for both inter-assay PCVs. Comparison studies using 50 patient and spiked serum samples showed r values of 0.9890 and 0.9853 and y-intercept values of -1.918 and -1.357, respectively compared to the HPLC reference method. All data points were <+/-15% of the mean. Linearity (r = 0.9952 (SMX) and 0.9954 (TMP)) was established from 12 to 400 μg/mL with a detection limit of 0.47 μg/mL, and 1.2 to 40 μg/mL with a detection limit of 0.06 μg/mL, for SMX and TMP, respectively. For either drug, no significant carryover was observed after samples at the upper limit of quantification. No interference was observed from any of the 77 drugs and respective metabolites tested.

CONCLUSION

A high-throughput SPE-MS/MS method for TMP/SMX quantification was developed. The <20 second analysis time is a significant improvement compared to traditional HPLC and LC-MS/MS methods, without sacrificing analytical performance.

Authors+Show Affiliations

Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.Toxicology and Drug Monitoring Laboratory, Division of Clinical Biochemistry and Immunology. Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.Toxicology and Drug Monitoring Laboratory, Division of Clinical Biochemistry and Immunology. Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.Toxicology and Drug Monitoring Laboratory, Division of Clinical Biochemistry and Immunology. Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.Toxicology and Drug Monitoring Laboratory, Division of Clinical Biochemistry and Immunology. Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.Toxicology and Drug Monitoring Laboratory, Division of Clinical Biochemistry and Immunology. Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32618880

Citation

Fatica, Erica, et al. "Quantification of Serum Sulfamethoxazole and Trimethoprim By Ultra-Fast SPE-MS/MS." Therapeutic Drug Monitoring, 2020.
Fatica E, Faber J, Gaffron C, et al. Quantification of Serum Sulfamethoxazole and Trimethoprim by Ultra-Fast SPE-MS/MS. Ther Drug Monit. 2020.
Fatica, E., Faber, J., Gaffron, C., Bjergum, M., Langman, L., & Jannetto, P. (2020). Quantification of Serum Sulfamethoxazole and Trimethoprim by Ultra-Fast SPE-MS/MS. Therapeutic Drug Monitoring. https://doi.org/10.1097/FTD.0000000000000785
Fatica E, et al. Quantification of Serum Sulfamethoxazole and Trimethoprim By Ultra-Fast SPE-MS/MS. Ther Drug Monit. 2020 Jun 29; PubMed PMID: 32618880.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantification of Serum Sulfamethoxazole and Trimethoprim by Ultra-Fast SPE-MS/MS. AU - Fatica,Erica, AU - Faber,Jennifer, AU - Gaffron,Christopher, AU - Bjergum,Matthew, AU - Langman,Loralie, AU - Jannetto,Paul, Y1 - 2020/06/29/ PY - 2020/7/4/entrez JF - Therapeutic drug monitoring JO - Ther Drug Monit N2 - BACKGROUND: The combination of trimethoprim (TMP) and sulfamethoxazole (SMX) is used to treat a number of bacterial infections. TMP/SMX concentrations in serum are conventionally monitored using high-performance liquid chromatography (HPLC) or liquid chromatography tandem mass spectrometry (LC-MS/MS). These methods require laborious manual extraction techniques and relatively long sample analysis times, necessitating the development of a simple, high-throughput method. A simple, high-throughput method to measure TMP/SMX using ultra-fast solid-phase extraction-tandem mass spectrometry (SPE-MS/MS) has been developed. METHODS: Calibration standards, quality control materials, and patient samples were precipitated with acetonitrile containing isotopically labeled internal standards. Samples were vortexed, centrifuged for 5 minutes at 2053×g, and the resulting supernatant diluted in aqueous mobile phase and injected onto the C18 SPE cartridge. MS/MS analysis was carried out by electrospray ionization in positive ion mode at a rate of <20 seconds per sample. A five-point linear 1/x calibration curve was used to calculate sample concentrations. RESULTS: The intra-assay precision coefficients of variation (PCVs) were <6% and <7% for SMX and TMP, respectively, and <10% for both inter-assay PCVs. Comparison studies using 50 patient and spiked serum samples showed r values of 0.9890 and 0.9853 and y-intercept values of -1.918 and -1.357, respectively compared to the HPLC reference method. All data points were <+/-15% of the mean. Linearity (r = 0.9952 (SMX) and 0.9954 (TMP)) was established from 12 to 400 μg/mL with a detection limit of 0.47 μg/mL, and 1.2 to 40 μg/mL with a detection limit of 0.06 μg/mL, for SMX and TMP, respectively. For either drug, no significant carryover was observed after samples at the upper limit of quantification. No interference was observed from any of the 77 drugs and respective metabolites tested. CONCLUSION: A high-throughput SPE-MS/MS method for TMP/SMX quantification was developed. The <20 second analysis time is a significant improvement compared to traditional HPLC and LC-MS/MS methods, without sacrificing analytical performance. SN - 1536-3694 UR - https://www.unboundmedicine.com/medline/citation/32618880/Quantification_of_Serum_Sulfamethoxazole_and_Trimethoprim_by_Ultra-Fast_SPE-MS/MS L2 - https://doi.org/10.1097/FTD.0000000000000785 DB - PRIME DP - Unbound Medicine ER -
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