Lambert Kristiansen Sørensen

Determination of camostat and its metabolites in human plasma – Preservation of samples and quantification by a validated UHPLC-MS/ MS method

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Determination of camostat and its metabolites in human plasma – Preservation of samples and quantification by a validated UHPLC-MS/ MS method. / Sørensen, Lambert Kristiansen; Hasselstrøm, Jørgen Bo; Damsgaard Gunst, Jesper; Søgaard, Ole Schmeltz; Kjolby, Mads.

In: Clinical Biochemistry, Vol. 96, 10.2021, p. 56-62.

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@article{3bb1fca89218459ba7ddcacdd01cadc5,
title = "Determination of camostat and its metabolites in human plasma – Preservation of samples and quantification by a validated UHPLC-MS/ MS method",
abstract = "Objectives: Camostat mesilate is a drug that is being repurposed for new applications such as that against COVID-19 and prostate cancer. This induces a need for the development of an analytical method for the quantification of camostat and its metabolites in plasma samples. Camostat is, however, very unstable in whole blood and plasma due to its two ester bonds. The molecule is readily hydrolysed by esterases to 4-(4-guanidinobenzoyloxy)phenylacetic acid (GBPA) and further to 4-guanidinobenzoic acid (GBA). For reliable quantification of camostat, a technique is required that can instantly inhibit esterases when blood samples are collected. Design and methods: An ultra-high-performance liquid chromatography-tandem mass spectrometry method (UHPLC-ESI-MS/MS) using stable isotopically labelled analogues as internal standards was developed and validated. Different esterase inhibitors were tested for their ability to stop the hydrolysis of camostat ester bonds. Results: Both diisopropylfluorophosphate (DFP) and paraoxon were discovered as efficient inhibitors of camostat metabolism at 10 mM concentrations. No significant changes in camostat and GBPA concentrations were observed in fluoride-citrate-DFP/paraoxon-preserved plasma after 24 h of storage at room temperature or 4 months of storage at −20 °C and −80 °C. The lower limits of quantification were 0.1 ng/mL for camostat and GBPA and 0.2 ng/mL for GBA. The mean true extraction recoveries were greater than 90%. The relative intra-laboratory reproducibility standard deviations were at a maximum of 8% at concentrations of 1–800 ng/mL. The trueness expressed as the relative bias of the test results was within ±3% at concentrations of 1–800 ng/mL. Conclusions: A methodology was developed that preserves camostat and GBPA in plasma samples and provides accurate and sensitive quantification of camostat, GBPA and GBA by UHPLC-MS/MS.",
keywords = "Camostat, COVID-19, Esterase inhibitors, FOY-251, FOY-305, GBA, GBPA, LC-MS/MS",
author = "S{\o}rensen, {Lambert Kristiansen} and Hasselstr{\o}m, {J{\o}rgen Bo} and {Damsgaard Gunst}, Jesper and S{\o}gaard, {Ole Schmeltz} and Mads Kjolby",
year = "2021",
month = oct,
doi = "10.1016/j.clinbiochem.2021.07.007",
language = "English",
volume = "96",
pages = "56--62",
journal = "Clinical Biochemistry",
issn = "0009-9120",
publisher = "Elsevier Inc.",

}

RIS

TY - JOUR

T1 - Determination of camostat and its metabolites in human plasma – Preservation of samples and quantification by a validated UHPLC-MS/ MS method

AU - Sørensen, Lambert Kristiansen

AU - Hasselstrøm, Jørgen Bo

AU - Damsgaard Gunst, Jesper

AU - Søgaard, Ole Schmeltz

AU - Kjolby, Mads

PY - 2021/10

Y1 - 2021/10

N2 - Objectives: Camostat mesilate is a drug that is being repurposed for new applications such as that against COVID-19 and prostate cancer. This induces a need for the development of an analytical method for the quantification of camostat and its metabolites in plasma samples. Camostat is, however, very unstable in whole blood and plasma due to its two ester bonds. The molecule is readily hydrolysed by esterases to 4-(4-guanidinobenzoyloxy)phenylacetic acid (GBPA) and further to 4-guanidinobenzoic acid (GBA). For reliable quantification of camostat, a technique is required that can instantly inhibit esterases when blood samples are collected. Design and methods: An ultra-high-performance liquid chromatography-tandem mass spectrometry method (UHPLC-ESI-MS/MS) using stable isotopically labelled analogues as internal standards was developed and validated. Different esterase inhibitors were tested for their ability to stop the hydrolysis of camostat ester bonds. Results: Both diisopropylfluorophosphate (DFP) and paraoxon were discovered as efficient inhibitors of camostat metabolism at 10 mM concentrations. No significant changes in camostat and GBPA concentrations were observed in fluoride-citrate-DFP/paraoxon-preserved plasma after 24 h of storage at room temperature or 4 months of storage at −20 °C and −80 °C. The lower limits of quantification were 0.1 ng/mL for camostat and GBPA and 0.2 ng/mL for GBA. The mean true extraction recoveries were greater than 90%. The relative intra-laboratory reproducibility standard deviations were at a maximum of 8% at concentrations of 1–800 ng/mL. The trueness expressed as the relative bias of the test results was within ±3% at concentrations of 1–800 ng/mL. Conclusions: A methodology was developed that preserves camostat and GBPA in plasma samples and provides accurate and sensitive quantification of camostat, GBPA and GBA by UHPLC-MS/MS.

AB - Objectives: Camostat mesilate is a drug that is being repurposed for new applications such as that against COVID-19 and prostate cancer. This induces a need for the development of an analytical method for the quantification of camostat and its metabolites in plasma samples. Camostat is, however, very unstable in whole blood and plasma due to its two ester bonds. The molecule is readily hydrolysed by esterases to 4-(4-guanidinobenzoyloxy)phenylacetic acid (GBPA) and further to 4-guanidinobenzoic acid (GBA). For reliable quantification of camostat, a technique is required that can instantly inhibit esterases when blood samples are collected. Design and methods: An ultra-high-performance liquid chromatography-tandem mass spectrometry method (UHPLC-ESI-MS/MS) using stable isotopically labelled analogues as internal standards was developed and validated. Different esterase inhibitors were tested for their ability to stop the hydrolysis of camostat ester bonds. Results: Both diisopropylfluorophosphate (DFP) and paraoxon were discovered as efficient inhibitors of camostat metabolism at 10 mM concentrations. No significant changes in camostat and GBPA concentrations were observed in fluoride-citrate-DFP/paraoxon-preserved plasma after 24 h of storage at room temperature or 4 months of storage at −20 °C and −80 °C. The lower limits of quantification were 0.1 ng/mL for camostat and GBPA and 0.2 ng/mL for GBA. The mean true extraction recoveries were greater than 90%. The relative intra-laboratory reproducibility standard deviations were at a maximum of 8% at concentrations of 1–800 ng/mL. The trueness expressed as the relative bias of the test results was within ±3% at concentrations of 1–800 ng/mL. Conclusions: A methodology was developed that preserves camostat and GBPA in plasma samples and provides accurate and sensitive quantification of camostat, GBPA and GBA by UHPLC-MS/MS.

KW - Camostat

KW - COVID-19

KW - Esterase inhibitors

KW - FOY-251

KW - FOY-305

KW - GBA

KW - GBPA

KW - LC-MS/MS

UR - http://www.scopus.com/inward/record.url?scp=85110647446&partnerID=8YFLogxK

U2 - 10.1016/j.clinbiochem.2021.07.007

DO - 10.1016/j.clinbiochem.2021.07.007

M3 - Journal article

C2 - 34252447

AN - SCOPUS:85110647446

VL - 96

SP - 56

EP - 62

JO - Clinical Biochemistry

JF - Clinical Biochemistry

SN - 0009-9120

ER -