Purification of Immunomodulator Fraction Components of Garlic (R10) by HPLC

Abstract

Background: It has been reported that garlic extract and its components show medicinal effects including immunomodulatory activities. We have isolated the immunomodulatory fraction (R10) previously. Objective: In this study we have proposed to purify the components of R10 using HPLC. Methods: Crude garlic extract purchased from Hamadan, Iran. R10 fraction including 10-50 KD molecules have been isolated using ultrafiltration. Further purification has been made using Vaydac 208Tpv10, a semi-preparative HPLC reversphase choromatography column. Tricine SDS-PAGE has been used to determine molecular weight of the samples. Results: 6 major peakes were obtained from HPLC of R10 at gradient of 0.25 % of buffer B/min through 60 minute. The molecular weight of 3 peaks (samples 0, 1 and 3) was 12 KD with tricine-SDSPAGE. Conclusion: Using C8 HPLC reversphase choromatography seems to be appropriate tool for purification of R10 components and the purified components at peak 0, 1 and 3 seems to be isotypic molecules.

Keywords

• garlic
• immunomodulator fraction
• hplc
• purification