Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation Treatment
(1) Background: Hydrogen sulfide (H<sub>2</sub>S) is a widely recognized gasotransmitter, with key roles in physiological and pathological processes. The accurate quantification of H<sub>2</sub>S and reactive sulfur species (RSS) may hold important implications for the diagno...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-05-01
|
| Series: | Antioxidants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-3921/11/5/939 |
| _version_ | 1827670698686939136 |
|---|---|
| author | Barbara Roda Nan Zhang Laura Gambari Brunella Grigolo Cristina Eller-Vainicher Luigi Gennari Alessandro Zappi Stefano Giordani Valentina Marassi Andrea Zattoni Pierluigi Reschiglian Francesco Grassi |
| author_facet | Barbara Roda Nan Zhang Laura Gambari Brunella Grigolo Cristina Eller-Vainicher Luigi Gennari Alessandro Zappi Stefano Giordani Valentina Marassi Andrea Zattoni Pierluigi Reschiglian Francesco Grassi |
| author_sort | Barbara Roda |
| collection | DOAJ |
| description | (1) Background: Hydrogen sulfide (H<sub>2</sub>S) is a widely recognized gasotransmitter, with key roles in physiological and pathological processes. The accurate quantification of H<sub>2</sub>S and reactive sulfur species (RSS) may hold important implications for the diagnosis and prognosis of diseases. However, H<sub>2</sub>S species quantification in biological matrices is still a challenge. Among the sulfide detection methods, monobromobimane (MBB) derivatization coupled with reversed phase high-performance liquid chromatography (RP-HPLC) is one of the most reported. However, it is characterized by a complex preparation and time-consuming process, which may alter the actual H<sub>2</sub>S level; moreover, a quantitative validation has still not been described. (2) Methods: We developed and validated an improved analytical protocol for the MBB RP-HPLC method. MBB concentration, temperature and sample handling were optimized, and the calibration method was validated using leave-one-out cross-validation and tested in a clinical setting. (3) Results: The method shows high sensitivity and allows the quantification of H<sub>2</sub>S species, with a limit of detection of 0.5 µM. Finally, it can be successfully applied in measurements of H<sub>2</sub>S levels in the serum of patients subjected to inhalation with vapors rich in H<sub>2</sub>S. (4) Conclusions: These data demonstrate that the proposed method is precise and reliable for measuring H<sub>2</sub>S species in biological matrices and can be used to provide key insights into the etiopathogenesis of several diseases and sulfur-based treatments. |
| first_indexed | 2024-03-10T03:26:43Z |
| format | Article |
| id | doaj.art-c5a22069e72b423bbdd8064477648af1 |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-10T03:26:43Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-c5a22069e72b423bbdd8064477648af12023-11-23T09:52:01ZengMDPI AGAntioxidants2076-39212022-05-0111593910.3390/antiox11050939Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation TreatmentBarbara Roda0Nan Zhang1Laura Gambari2Brunella Grigolo3Cristina Eller-Vainicher4Luigi Gennari5Alessandro Zappi6Stefano Giordani7Valentina Marassi8Andrea Zattoni9Pierluigi Reschiglian10Francesco Grassi11Department of Chemistry “G. Ciamician”, University of Bologna, 40126 Bologna, ItalyDepartment of Chemistry “G. Ciamician”, University of Bologna, 40126 Bologna, ItalyLaboratorio RAMSES, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, ItalyLaboratorio RAMSES, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, ItalyUnit of Endocrinology, Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Ca’ Granda-Ospedale Maggiore Policlinico, 20122 Milan, ItalyDepartment of Medicine, Surgery and Neurosciences, University of Siena, 53100 Siena, ItalyDepartment of Chemistry “G. Ciamician”, University of Bologna, 40126 Bologna, ItalyDepartment of Chemistry “G. Ciamician”, University of Bologna, 40126 Bologna, ItalyDepartment of Chemistry “G. Ciamician”, University of Bologna, 40126 Bologna, ItalyDepartment of Chemistry “G. Ciamician”, University of Bologna, 40126 Bologna, ItalyDepartment of Chemistry “G. Ciamician”, University of Bologna, 40126 Bologna, ItalyLaboratorio RAMSES, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, Italy(1) Background: Hydrogen sulfide (H<sub>2</sub>S) is a widely recognized gasotransmitter, with key roles in physiological and pathological processes. The accurate quantification of H<sub>2</sub>S and reactive sulfur species (RSS) may hold important implications for the diagnosis and prognosis of diseases. However, H<sub>2</sub>S species quantification in biological matrices is still a challenge. Among the sulfide detection methods, monobromobimane (MBB) derivatization coupled with reversed phase high-performance liquid chromatography (RP-HPLC) is one of the most reported. However, it is characterized by a complex preparation and time-consuming process, which may alter the actual H<sub>2</sub>S level; moreover, a quantitative validation has still not been described. (2) Methods: We developed and validated an improved analytical protocol for the MBB RP-HPLC method. MBB concentration, temperature and sample handling were optimized, and the calibration method was validated using leave-one-out cross-validation and tested in a clinical setting. (3) Results: The method shows high sensitivity and allows the quantification of H<sub>2</sub>S species, with a limit of detection of 0.5 µM. Finally, it can be successfully applied in measurements of H<sub>2</sub>S levels in the serum of patients subjected to inhalation with vapors rich in H<sub>2</sub>S. (4) Conclusions: These data demonstrate that the proposed method is precise and reliable for measuring H<sub>2</sub>S species in biological matrices and can be used to provide key insights into the etiopathogenesis of several diseases and sulfur-based treatments.https://www.mdpi.com/2076-3921/11/5/939hydrogen sulfide poolbiomarkersbone metabolismhigh-performance liquid chromatography with fluorescencemonobromobimanesulfur species |
| spellingShingle | Barbara Roda Nan Zhang Laura Gambari Brunella Grigolo Cristina Eller-Vainicher Luigi Gennari Alessandro Zappi Stefano Giordani Valentina Marassi Andrea Zattoni Pierluigi Reschiglian Francesco Grassi Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation Treatment Antioxidants hydrogen sulfide pool biomarkers bone metabolism high-performance liquid chromatography with fluorescence monobromobimane sulfur species |
| title | Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation Treatment |
| title_full | Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation Treatment |
| title_fullStr | Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation Treatment |
| title_full_unstemmed | Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation Treatment |
| title_short | Optimization of a Monobromobimane (MBB) Derivatization and RP-HPLC-FLD Detection Method for Sulfur Species Measurement in Human Serum after Sulfur Inhalation Treatment |
| title_sort | optimization of a monobromobimane mbb derivatization and rp hplc fld detection method for sulfur species measurement in human serum after sulfur inhalation treatment |
| topic | hydrogen sulfide pool biomarkers bone metabolism high-performance liquid chromatography with fluorescence monobromobimane sulfur species |
| url | https://www.mdpi.com/2076-3921/11/5/939 |
| work_keys_str_mv | AT barbararoda optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT nanzhang optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT lauragambari optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT brunellagrigolo optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT cristinaellervainicher optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT luigigennari optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT alessandrozappi optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT stefanogiordani optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT valentinamarassi optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT andreazattoni optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT pierluigireschiglian optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment AT francescograssi optimizationofamonobromobimanembbderivatizationandrphplcflddetectionmethodforsulfurspeciesmeasurementinhumanserumaftersulfurinhalationtreatment |