Quantification of thyrotropin-releasing hormone by liquid chromatography-electrospray mass spectrometry.
Thyrotropin-releasing hormone (TRH) is involved in a wide range of biological responses. It has a central role in the endocrine system and regulates several neurobiological activities. In the present study, a rapid, sensitive and selective liquid chromatography-mass spectrometry method for the identification and quantification of TRH has been developed. The methodology takes advantage of the specificity of the selected-ion monitoring acquisition mode with a limit of detection of 1 fmol. Furthermore, the MS/MS fragmentation pattern of TRH has been investigated to develop a selected reaction monitoring (SRM) method that allows the detection of a specific b2 product ion at m/z 249.1, corresponding to the N-terminus dipeptide pyroglutamic acid-histidine. The method has been tested on rat hypothalami to evaluate its suitability for the detection within very complex biological samples.
Chambery
Angela
A
Dipartimento di Scienze della Vita, Seconda Università di Napoli, Via Vivaldi 43, 81100, Caserta, Italy. angela.chambery@unina2.it
Severino
Valeria
V
Di Maro
Antimo
A
D'Aniello
Antimo
A
Ruvo
Menotti
M
Parente
Augusto
A
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004151:Dipeptides / Q000032:analysis / Q000737:chemistry; D007031:Hypothalamus / Q000737:chemistry; D057230:Limit of Detection; D008832:Microchemistry / Q000379:methods; D015394:Molecular Structure; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives / Q000032:analysis / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D013973:Thyrotropin-Releasing Hormone / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification
D000596:Amino Acids; D004151:Dipeptides; C058050:pyroglutamylhistidine; D013973:Thyrotropin-Releasing Hormone; D011761:Pyrrolidonecarboxylic Acid