Determination Of4Phytohormones Of Latex Of Hevea Brasiliensis By HPLC

The plant hormones Zeatin (ZT), Gibberellin (GA), Auxin (IAA) and Abscisic acid (ABA) in rubber (Hevea brasiliensis) latex had been isolated and purified, then determined qualitatively and quantitatively by high performance liquid chromatography (HPLC). A new HPLC method for isolation and measurement of the above phytohormones was developed.A new method of extraction and isolation of4phytohormones including GA, IAA, ABA and IPA from latex was developed. Phytohormones were isolated by that the frest rubber latex was extracted with80%ice-cold methanol extraction, then centrifugating, and then the supernatant from centrifugation passed through C18pre-column, and extracted with ethyl acetate after adjusting pH of the colatuie, and then purified by high performance liquid chromatography. Four phytohormones were detected with isocratic elution, methanol, acetonitrile and0.5%acetic acid (20/20/60v/v) as mobile phase. Any of the4phytohormones could not be detected with other methods of plant tissue phytohormones extraction,.A new method of extraction and isolation of4phytohormones, including ZT, GA, IAA and ABA from rubber latex was developed. Phytohormones were isolated by that the frest rubber latex extracted with80%ice-cold methanol extraction, then centrifugating, then the supernatant from centrifugation was evaporated to the aqueous phase under vacuum condition. After adjusted pH value, the extraction was further purified through PXC pre-column. GA, IAA and ABA were eluted followed by ZT. Then detected by HPLC methods.A new elution method of4phytohormones, including GA, IAA, ABA and ZT from rubber latex was developed. The ZT was detected, peak time was5.426min, recovery rate was87%, with isocratic elution, the mobile phase was methanol and0.5%acetic acid (20/80v/v), the analysis time was10min. The GA, IAA, and ABA were detected, the peak time of GA, IAA, and ABA was13.377,16.581,and18.988 min and the recovery rate was70%,82%, and92%respectively, with gradient elution, mobile phase was methanol and0.5%acetic acid (20/80v/v), the analysis time was34min. It would gave a good result of isolation and detection of4phytohormones due to the concentration of impurities were reduced by that the same sample was detected separatedly.The results showed that these methods gave a high reproducibility and operated simplely that was suitable for the study of isolation and detection of phytohormones in rubber latex.