Production of microalgae-derived extract and its respective HPLC analysis / Amir Bakhtiar Abdul Razak

Microalgae have great potential in biopharrnaceutical or medical application as they carries a lot of promising bioactive compounds such as carotenoids, vitamins and polyunsaturated fatty acids (PUF As). Due to that, further study in Microalgae Research Laboratory (MRL) was conducted in order to develop a method to extract these microalgae compounds and generate the HPLC chromatograms to analyze these microalgae-derived extract (MDE). Two strains of microalgae in MRL were chosen based on the availability of their growth profiles from previous study and these microalgae were known as ZS and Ku Lorn 2 X2. These microalgae were subjected to the solvent extraction. The solvents used were hexane, ethyl acetate and 80°C deionised water. Solvents with different polarities were chosen because polarity greatly affects the extraction process. Polar solvent can extract the polar compounds and non-polar solvent can extract the non-polar compounds. In this study, HPLC was used to generate the MDE chromatogram. The standard operating procedure (SOP) developed at Atta-ur-Rahrnan Research Institute for Natural Product Discovery (AURiND) was used and the column used was Phenomenex Cl8 Hydro 4 µm, 4.6 x 150 mm. Two mobile phase used were ultrapure water as mobile phase A and acetonitrile (ACN) as mobile phase B. Flow rate was maintained at 1 mL/min and each samples were ran for 30 min with post time of 5 min. The detectors used were diode array detector (DAD) and evaporative light scattering detector (ELSD). Hexane and ethyl acetate was capable of extracting these microalgae compounds since there were significant peaks shown in the MDE chromatograms. However, there was no significant peak shown in chromatograms generated from 80°C deionised water extract. The identities of the MDE are still unknown due to the unavailability of standard in this study. The standard need to be design so that by comparing the retention time (tR), the identities as well as quantities of the compounds can be determined.