A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing Immunotherapy
Tryptophan (TRP) is an essential amino acid crucial for the production of many bioactive compounds. Disturbances in TRP metabolism have been revealed in various diseases, many of which are closely related to the immune system. In recent years, we have focused on finding blood-based biomarkers of suc...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-12-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/30/1/121 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841549062928596992 |
|---|---|
| author | Anna Siemiątkowska Barbara Kuźnar-Kamińska Katarzyna Kosicka-Noworzyń Kamila Nowaczewska Hanna Winiarska Dominika Popiołek Filip Kamiński Franciszek K. Główka |
| author_facet | Anna Siemiątkowska Barbara Kuźnar-Kamińska Katarzyna Kosicka-Noworzyń Kamila Nowaczewska Hanna Winiarska Dominika Popiołek Filip Kamiński Franciszek K. Główka |
| author_sort | Anna Siemiątkowska |
| collection | DOAJ |
| description | Tryptophan (TRP) is an essential amino acid crucial for the production of many bioactive compounds. Disturbances in TRP metabolism have been revealed in various diseases, many of which are closely related to the immune system. In recent years, we have focused on finding blood-based biomarkers of successful immunotherapy in cancer. Thus, we aimed to develop a robust liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for TRP and its metabolites that could be used in biomarker studies. Although analyzing TRP derivatives in biological matrices is not a new topic, we encountered multiple challenges during method development. One of them was the phenomenon of cross-interferences between the analyzed molecules, which has not been explored in most published papers. We noticed that injecting a pure single-compound solution often generated a signal in the other compounds’ MS/MS channels. Specifically, TRP generated unexpected peaks in the channel for kynurenine, kynurenic acid, and xanthurenic acid, while kynurenine generated peaks in the channel for kynurenic acid. We also recorded a mutual cross-talk between kynurenine and isotope-labeled TRP. Different origins of the observed cross-signal contribution were proposed. This paper draws attention to investigating cross-interferences in LC-MS/MS, especially when structurally related compounds will be analyzed. Despite all the challenges, the method was successfully validated according to international guidelines (EMA/ICH), and its applicability was confirmed in a pilot study including 20 patients with lung cancer undergoing chemoimmunotherapy. |
| format | Article |
| id | doaj-art-42828e2296c84b7d9432bbb350eee35d |
| institution | Kabale University |
| issn | 1420-3049 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj-art-42828e2296c84b7d9432bbb350eee35d2025-01-10T13:18:56ZengMDPI AGMolecules1420-30492024-12-0130112110.3390/molecules30010121A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing ImmunotherapyAnna Siemiątkowska0Barbara Kuźnar-Kamińska1Katarzyna Kosicka-Noworzyń2Kamila Nowaczewska3Hanna Winiarska4Dominika Popiołek5Filip Kamiński6Franciszek K. Główka7Department of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 3 Rokietnicka Street, 60-806 Poznań, PolandDepartment of Pulmonology, Allergology and Respiratory Oncology, Poznan University of Medical Sciences, 84 Szamarzewskiego Street, 60-569 Poznań, PolandDepartment of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 3 Rokietnicka Street, 60-806 Poznań, PolandDepartment of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 3 Rokietnicka Street, 60-806 Poznań, PolandDepartment of Pulmonology, Allergology and Respiratory Oncology, Poznan University of Medical Sciences, 84 Szamarzewskiego Street, 60-569 Poznań, PolandDepartment of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 3 Rokietnicka Street, 60-806 Poznań, PolandDepartment of Pulmonology, Allergology and Respiratory Oncology, Poznan University of Medical Sciences, 84 Szamarzewskiego Street, 60-569 Poznań, PolandDepartment of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 3 Rokietnicka Street, 60-806 Poznań, PolandTryptophan (TRP) is an essential amino acid crucial for the production of many bioactive compounds. Disturbances in TRP metabolism have been revealed in various diseases, many of which are closely related to the immune system. In recent years, we have focused on finding blood-based biomarkers of successful immunotherapy in cancer. Thus, we aimed to develop a robust liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for TRP and its metabolites that could be used in biomarker studies. Although analyzing TRP derivatives in biological matrices is not a new topic, we encountered multiple challenges during method development. One of them was the phenomenon of cross-interferences between the analyzed molecules, which has not been explored in most published papers. We noticed that injecting a pure single-compound solution often generated a signal in the other compounds’ MS/MS channels. Specifically, TRP generated unexpected peaks in the channel for kynurenine, kynurenic acid, and xanthurenic acid, while kynurenine generated peaks in the channel for kynurenic acid. We also recorded a mutual cross-talk between kynurenine and isotope-labeled TRP. Different origins of the observed cross-signal contribution were proposed. This paper draws attention to investigating cross-interferences in LC-MS/MS, especially when structurally related compounds will be analyzed. Despite all the challenges, the method was successfully validated according to international guidelines (EMA/ICH), and its applicability was confirmed in a pilot study including 20 patients with lung cancer undergoing chemoimmunotherapy.https://www.mdpi.com/1420-3049/30/1/121cross-signal contributioncross-interferencescross-talkstandard impuritysimilar MRMspoor analyte stability |
| spellingShingle | Anna Siemiątkowska Barbara Kuźnar-Kamińska Katarzyna Kosicka-Noworzyń Kamila Nowaczewska Hanna Winiarska Dominika Popiołek Filip Kamiński Franciszek K. Główka A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing Immunotherapy Molecules cross-signal contribution cross-interferences cross-talk standard impurity similar MRMs poor analyte stability |
| title | A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing Immunotherapy |
| title_full | A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing Immunotherapy |
| title_fullStr | A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing Immunotherapy |
| title_full_unstemmed | A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing Immunotherapy |
| title_short | A Novel Liquid Chromatography–Tandem Mass Spectrometry Method to Quantify Tryptophan and Its Major Metabolites in Serum to Support Biomarker Studies in Patients with Cancer Undergoing Immunotherapy |
| title_sort | novel liquid chromatography tandem mass spectrometry method to quantify tryptophan and its major metabolites in serum to support biomarker studies in patients with cancer undergoing immunotherapy |
| topic | cross-signal contribution cross-interferences cross-talk standard impurity similar MRMs poor analyte stability |
| url | https://www.mdpi.com/1420-3049/30/1/121 |
| work_keys_str_mv | AT annasiemiatkowska anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT barbarakuznarkaminska anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT katarzynakosickanoworzyn anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT kamilanowaczewska anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT hannawiniarska anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT dominikapopiołek anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT filipkaminski anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT franciszekkgłowka anovelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT annasiemiatkowska novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT barbarakuznarkaminska novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT katarzynakosickanoworzyn novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT kamilanowaczewska novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT hannawiniarska novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT dominikapopiołek novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT filipkaminski novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy AT franciszekkgłowka novelliquidchromatographytandemmassspectrometrymethodtoquantifytryptophananditsmajormetabolitesinserumtosupportbiomarkerstudiesinpatientswithcancerundergoingimmunotherapy |