Purification of immunomodulator fraction components of garlic [R10] by HPLC

ABSTRACT

It has been reported that garlic extract and its components show medicinal effects including immunomodulatory activities. We have isolated the immunomodulatory fraction [R10] previously In this study we have proposed to purify the components of R10 using HPLC. 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. 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. 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