Affiliations: [a] Department of Clinical Pharmacy, College of Pharmacy, King Khalid University, Abha, Saudi Arabia
| [b] Pharmaceutical Science, Oregon State University, Corvallis, OR, USA
Correspondence:
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Corresponding author: Sultan M. Alshahrani, Department of Clinical Pharmacy, King Khalid University, Asir, Abha, Kingdom of Saudi Arabia. Phone: +00966 508747473; E-mail: [email protected].
Abstract: This study was designed to develop and validate a simple and efficient high performance liquid chromatography (HPLC) method to determine flunixin concentrations in Asian elephant’s (Elephas maximus) plasma. Flunixin was administered orally at a dose of 0.8 mg/kg, and blood samples were collected. Flunixin extraction was performed by adding an equal amount of acetonitrile to plasma and centrifuging at 4500 rpm for 25 minutes. The supernatant was removed, and flunixin was analyzed using HPLC-UV detection. Two methods were developed and tested utilizing two different mobile phases either with or without adding methanol (ACN: H2O vs. ACN: H2O: MeOH). Both methods showed excellent linearity and reproducibility. The limit of detection was 0.05 ug/ml and limit of quantification was 0.1 ug/ml. the efficiency of flunixin recovery was maximized by the addition of methanol to mobile phase (ACN: H2O: MeOH as 50:30:20) at 95% in comparison to 23% without methanol. In conclusion, adding methanol to HPLC methods for extraction of flunixin from elephants’ plasma yielded higher recovery rate than without methanol.
Keywords: Flunixin, Elephas maximus, linearity and reproducibility, total quality control
