HPLC Simultaneous Determination of Five Nucleoside Compounds in Extracts of Buthus martensii by Different Extraction Technologies

Abstract:

Objective: To compare effects of 3 extraction technologies (water boiling,alcohol extraction and wet ultrafine grinding) on the content of uracil,hypoxanthine,xanthine,guanosine and adenosine from Buthus martensii. Method: Samples of B. martensii were extracted with different technologies,the content of five nucleosides was determined by HPLC.Conditions were as followings:Aglient Zorbax C₁₈ column (4.6 mm×250 mm,5 μm),gradient eluted with 0.07% ice acetic acid-methanol as mobile phase,flow rate 1.0 mL·min^-1,detection wavelength 254 nm and column temperature 35 ℃. Result: Uracil, hypoxanthine, xanthine, guanosine, adenosine had a good linear relationship in a certain range of concentration,r>0.999 5,the average recoveries of five nucleoside compounds in all samples were between 98.9%-103.9% with RSD<3%. Conclusion: This established HPLC was convenient and accurate with good repeatability,it could be used to determine simultaneously the content of five nucleosides(uracil, hypoxanthine, xanthine, guanosine, adenosine) and compare difference among extracts of B. martensii by different technologies.