VENOM ANALYSIS RESULTS

Sample sentby: Sanlıurfa Akrep ve Yılan Zehiri Üretimi İlaç San Tic. Ltd. Şti.

Sample Delivery Date: 25.04.2024

Sample Information: Scorpion Venom, Androctonus turkiyensis (Species determination was made by Prof. Dr. Ersen Aydın YAĞMUR), Supplied in liquid form, white color.

Sample Purity: %100

Analysis Date: 25.04.2024

Analyzes Performed: UV-visibleSpectrophotometricAnalysis,SDS-PAGEandHPLC

Preparation Studies

Venom belonging to the Androctonus turkiyensis scorpion species, supplied in liquid white form,wasfirstpurifiedwithsaline-containingwater.Inordertoremoveexcesswaterfromthe venom structure, it was subjected to lyophilization process. Pure white powdered venom was obtained.

SpectrumResults

SpectraanalysisofvenomobtainedfromAndroctonusturkiyensisspeciesscorpionpreparedas   1

mg/mL was performed using Shimadzu brand UV-1900i model spectrophotometer device. Spectrum scanning was performed between 190 nm - 400 nm wavelengths. A venom-specific maximum peak was observed at 280 nm (Figure 1). For this reason, HPLC analyzes were measured based on 280 nm.

Figure1SpectrumanalysisofAndroctonusturkiyensis venom

HPLC Analysis

Powdered venoms were loaded into the HPLC (High Performance Liquid Chromatography) instrument at a concentration of 1000 ppm. In Shimadzu Prominence-i LC2030 3D HPLC device,SolventBwasmadeina0-60%lowgradientprogramwithaflowrateof1mg/mL.The spectrumwastakentodeterminethemaximumabsorbanceoftheproteinsinthevenom.Inline withtheseresults,HPLCanalysiswasperformedat280nmwavelength.Proteinsinthevenom       were detected in fractions at different times (Figure 2). As a result of HPLC analysis, 20 differentproteins/peptidesweredeterminedinthevenomofAndroctonusturkiyensis.The retention time of the fractions was indicated (Table 1).

Figure2.HPLCAnalysisresultofAndroctonusturkiyensis venom.

Table1.RetentiontimesoffractionsofAndroctonusturkiyensis venom