Quantitation of endogenous GnRH by validated ...
Document type :
Article dans une revue scientifique: Article original
PMID :
Permalink :
Title :
Quantitation of endogenous GnRH by validated nano-HPLC-HRMS method: a pilot study on ewe plasma
Author(s) :
Mecarelli, E. [Auteur]
Aigotti, R. [Auteur]
Asteggiano, A. [Auteur]
Giacobini, Paolo [Auteur]
Lille Neurosciences & Cognition (LilNCog) - U 1172
Chasles, Manon [Auteur]
Physiologie de la reproduction et des comportements [Nouzilly] [PRC]
Tillet, Yves [Auteur]
Dal Bello, F. [Auteur]
Medana, C. [Auteur]
Aigotti, R. [Auteur]
Asteggiano, A. [Auteur]
Giacobini, Paolo [Auteur]

Lille Neurosciences & Cognition (LilNCog) - U 1172
Chasles, Manon [Auteur]
Physiologie de la reproduction et des comportements [Nouzilly] [PRC]
Tillet, Yves [Auteur]
Dal Bello, F. [Auteur]
Medana, C. [Auteur]
Journal title :
Analytical and Bioanalytical Chemistry
Volume number :
414
Pages :
7623–7634
Publication date :
2022-09-11
ISSN :
1618-2642
Keyword(s) :
GnRH
PNA
Nano-HPLC-HRMS
Method validation
Surrogate matrix
Endogenous peptide
PNA
Nano-HPLC-HRMS
Method validation
Surrogate matrix
Endogenous peptide
HAL domain(s) :
Sciences du Vivant [q-bio]
English abstract : [en]
Gonadotropin-releasing hormone isoform I (GnRH), a neuro-deca-peptide, plays a fundamental role in development and maintenance of the reproductive system in vertebrates. The anomalous release of GnRH is observed in ...
Show more >Gonadotropin-releasing hormone isoform I (GnRH), a neuro-deca-peptide, plays a fundamental role in development and maintenance of the reproductive system in vertebrates. The anomalous release of GnRH is observed in reproductive disorder such as hypogonadotropic hypogonadism, polycystic ovary syndrome (PCOS), or following prenatal exposure to elevated androgen levels. Quantitation of GnRH plasma levels could help to diagnose and better understand these pathologies. Here, a validated nano-high-performance liquid chromatography–high-resolution mass spectrometry (HPLC-HRMS) method to quantify GnRH in ewe plasma samples is presented. Protein precipitation and solid-phase extraction (SPE) pre-treatment steps were required to purify and enrich GnRH and internal standard (lamprey-luteinizing hormone-releasing hormone-III, l-LHRH-III). For the validation process, a surrogate matrix approach was chosen following the International Council for Harmonisation (ICH) and FDA guidelines. Before the validation study, the validation model using the surrogate matrix was compared with those using a real matrix such as human plasma. All the tested parameters were analogous confirming the use of the surrogate matrix as a standard calibration medium. From the validation study, limit of detection (LOD) and limit of quantitation (LOQ) values of 0.008 and 0.024 ng/mL were obtained, respectively. Selectivity, accuracy, precision, recovery, and matrix effect were assessed with quality control samples in human plasma and all values were acceptable. Sixteen samples belonging to healthy and prenatal androgen (PNA) exposed ewes were collected and analyzed, and the GnRH levels ranged between 0.05 and 3.26 ng/mL. The nano-HPLC-HRMS developed here was successful in measuring GnRH, representing therefore a suitable technique to quantify GnRH in ewe plasma and to detect it in other matrices and species.Show less >
Show more >Gonadotropin-releasing hormone isoform I (GnRH), a neuro-deca-peptide, plays a fundamental role in development and maintenance of the reproductive system in vertebrates. The anomalous release of GnRH is observed in reproductive disorder such as hypogonadotropic hypogonadism, polycystic ovary syndrome (PCOS), or following prenatal exposure to elevated androgen levels. Quantitation of GnRH plasma levels could help to diagnose and better understand these pathologies. Here, a validated nano-high-performance liquid chromatography–high-resolution mass spectrometry (HPLC-HRMS) method to quantify GnRH in ewe plasma samples is presented. Protein precipitation and solid-phase extraction (SPE) pre-treatment steps were required to purify and enrich GnRH and internal standard (lamprey-luteinizing hormone-releasing hormone-III, l-LHRH-III). For the validation process, a surrogate matrix approach was chosen following the International Council for Harmonisation (ICH) and FDA guidelines. Before the validation study, the validation model using the surrogate matrix was compared with those using a real matrix such as human plasma. All the tested parameters were analogous confirming the use of the surrogate matrix as a standard calibration medium. From the validation study, limit of detection (LOD) and limit of quantitation (LOQ) values of 0.008 and 0.024 ng/mL were obtained, respectively. Selectivity, accuracy, precision, recovery, and matrix effect were assessed with quality control samples in human plasma and all values were acceptable. Sixteen samples belonging to healthy and prenatal androgen (PNA) exposed ewes were collected and analyzed, and the GnRH levels ranged between 0.05 and 3.26 ng/mL. The nano-HPLC-HRMS developed here was successful in measuring GnRH, representing therefore a suitable technique to quantify GnRH in ewe plasma and to detect it in other matrices and species.Show less >
Language :
Anglais
Audience :
Internationale
Popular science :
Non
Administrative institution(s) :
Université de Lille
Inserm
CHU Lille
Inserm
CHU Lille
Collections :
Submission date :
2024-01-16T00:49:19Z
2024-07-01T08:03:40Z
2024-07-01T08:03:40Z
Files
- s00216-022-04293-z.pdf
- Non spécifié
- Open access
- Access the document