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

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

[ cytotoxic effects a of ACA1 on human melanoma cell line]

Different methods such as surgery, chemotherapy, radiotherapy, hormone therapy and immunotherapy are used for treatment of melanoma cancer. Unfortunately they don't always have desirable results and they may have unfavorable side effects. Researchers try to find new, more effective drugs with low side effects. In this study we evaluated the cytotoxic effect of ACA-1, a water extract of a traditional Iranian medicinal herbs on a melanoma cell line SKMEL-3. The SKMEL3 cell line was obtained from Pasture institute, Tehran, Iran and cultured in RPMI media supplemented with 10% FBS. Equal number of cells were added to a 96 well microplate and were incubated with various doses of ACA1 [5,2,1,0.2,0.1,0.05,0.02 and 0.01 mg/ml] for 24, 48 and 72 hours in parallel. The cytotoxic effects of the drug was evaluated using MTT assay. The Results showed that ACA1 has significant cytotoxic effects with dose and time dependent manner on SKMEL3. The optimum dose [5 mg/ml] showed 47% cytotoxicity in 24 h, 65% cytotoxicity in 48 h and 71% cytotoxicity in 72 h. Based on the results of this research, ACA1 is a suitable candidate for chemotherapy of melanoma patients. Further studies are necessary in order to find effective drugs, their effects on other cell lines and approved in vivo models and clinical trials