Development of a sensitive and quantitative HPLC-FLD method for the determination of obestatin in human plasma

Karaca S., TEMEL T., YENİCELİ UĞUR D.

JOURNAL OF PEPTIDE SCIENCE, cilt.29, sa.11, 2023 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 29 Sayı: 11
  • Basım Tarihi: 2023
  • Doi Numarası: 10.1002/psc.3511
  • Dergi Adı: JOURNAL OF PEPTIDE SCIENCE
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, INSPEC, MEDLINE, Veterinary Science Database
  • Anahtar Kelimeler: chromatography, experimental design, obestatin, peptide, quantification
  • Anadolu Üniversitesi Adresli: Evet

Özet

Obestatin is a gastrointestinal system peptide. The quantification of this peptide is conventionally performed using immunological techniques. In this study, a selective and sensitive HPLC method coupled with fluorescence detection for the quantitation of obestatin in human plasma was developed and validated. The separation was obtained on a C-18 (4.6 x 100 mm, 3.5-mu m particles) column using a mobile phase composed of acetonitrile and water, both including 0.1% trifluoroacetic acid. The developed method was found to be linear in the concentration range of 20 to 1000 ng/mL, with a coefficient of determination of 0.9982. The precision results were less than 10%, and the accuracy results were between 92% and 107%. The detection and quantification limit values were obtained as 2.8 and 9.4 ng/mL, respectively. Analyte solutions were found stable for 24 h at room temperature, three freeze-thaw cycles, and 2 weeks at -20 degrees C. The developed method was successfully used for the quantification of obestatin in human plasma samples. In conclusion, the developed method is sensitive and specific for measuring the plasma concentrations of obestatin.