دورية أكاديمية
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LC-MS/MS Analysis Elucidates a Daily Rhythm in Orexin A Concentration in the Rat Vitreous Body.

التفاصيل البيبلوغرافية
العنوان: LC-MS/MS Analysis Elucidates a Daily Rhythm in Orexin A Concentration in the Rat Vitreous Body.
المؤلفون: Chrobok L; Department of Neurophysiology and Chronobiology, Institute of Zoology and Biomedical Research, Jagiellonian University in Krakow, Gronostajowa 9 Street, 30-387 Krakow, Poland., Bajkacz S; Department of Inorganic Chemistry, Analytical Chemistry and Electrochemistry, Faculty of Chemistry, Silesian University of Technology, Krzywoustego 6 Street, 44-100 Gliwice, Poland.; The Biotechnology Centre, Silesian University of Technology, Krzywoustego 8 Street, 44-100 Gliwice, Poland., Klich JD; Department of Neurophysiology and Chronobiology, Institute of Zoology and Biomedical Research, Jagiellonian University in Krakow, Gronostajowa 9 Street, 30-387 Krakow, Poland., Lewandowski MH; Department of Neurophysiology and Chronobiology, Institute of Zoology and Biomedical Research, Jagiellonian University in Krakow, Gronostajowa 9 Street, 30-387 Krakow, Poland.
المصدر: Molecules (Basel, Switzerland) [Molecules] 2021 Aug 19; Vol. 26 (16). Date of Electronic Publication: 2021 Aug 19.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: MDPI Country of Publication: Switzerland NLM ID: 100964009 Publication Model: Electronic Cited Medium: Internet ISSN: 1420-3049 (Electronic) Linking ISSN: 14203049 NLM ISO Abbreviation: Molecules
أسماء مطبوعة: Original Publication: Basel, Switzerland : MDPI, c1995-
مواضيع طبية MeSH: Orexins*/metabolism , Tandem Mass Spectrometry*/methods , Vitreous Body*/metabolism, Animals ; Rats ; Chromatography, Liquid/methods ; Circadian Rhythm/physiology ; Male ; Chromatography, High Pressure Liquid/methods ; Liquid Chromatography-Mass Spectrometry
مستخلص: Orexins are two neuropeptides synthesised mainly in the brain lateral hypothalamic area. The orexinergic system provides arousal-dependent cues for a plethora of brain centres, playing a vital role in feeding behaviour, regulation of the sleep-wake cycle and circadian rhythms. Recently, orexins were found to be produced in the retina of an eye; however, their content in the vitreous body and possible daily pattern of expression have not yet been explored. In this manuscript, we describe the development and validation of a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method designed for quantitative bioanalysis of orexin in the rat vitreous body. Orexin was extracted from vitreous body samples with a water:acetonitrile:formic acid (80:20:0.1; v / v / v ) mixture followed by vortexing and centrifuging. Separation was performed on a reverse-phase HPLC column under gradient conditions. Orexin was analysed via multiple-reaction monitoring (MRM) in the positive electrospray mode. The total analysis time for each sample was less than 5.0 min. Once the method was fully optimised, it was then validated, following the 2018 FDA guidance on bioanalytical method validations. The calibration curves for orexin (1-500 ng/mL) were constructed using a linear regression with a 1/x 2 weighting. The lower limit of quantitation for orexin was 1.0 pg/mL for the vitreous body. Intra-day and inter-day estimates of accuracy and precision were within 10% of their nominal values, indicating that the method is reliable for quantitation of orexin in the rat vitreous body. From the physiological perspective, our results are the first to show daily rhythm of orexin synthesis by the retina with possible implications on the circadian regulation of vision.
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معلومات مُعتمدة: 'Sonatina 2' 2018/28/C/NZ4/00099 National Science Centre
فهرسة مساهمة: Keywords: LC-MS/MS; circadian clock; orexin; vitreous body
المشرفين على المادة: 0 (Orexins)
تواريخ الأحداث: Date Created: 20210827 Date Completed: 20240725 Latest Revision: 20240725
رمز التحديث: 20240726
مُعرف محوري في PubMed: PMC8401233
DOI: 10.3390/molecules26165036
PMID: 34443622
قاعدة البيانات: MEDLINE
الوصف
تدمد:1420-3049
DOI:10.3390/molecules26165036