Physalis Minima Lab Report

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CHAPTER III

RESEARCH METHOLOGY

1.0 Plant extraction of Physalis minima

1.1 Preparation of fresh sample extract
Physalis minima L. at maturity state was harvested from Kepala Batas, identified and washed with a distilled water. The dried specimen was submitted to the USM Herbarium for future reference. About 200 g weight of leaves were blended finely with 300 mL distilled water using a blender until become a mixture with a small size of leaves. The water mixture of leaves were macerated with 1200 mL methanol and shaken continuously at 250 rpm for 24 hours. The mixtures was filtered with Whatman #1 filter paper and the filtrate was used to repeat the extraction for another two times. Then, the methanol solvent in mixtures was removed using the Rotary Evaporator (EYELA, N1100) at 140° hPa; 60°C; speed 5. The distilled water and excessive methanol were removed with Freeze Dryer (GENEVAC LTO, EZ 2.3 ELITE) for a week. The crude obtained were weighed and stored at -20°C until use.

1.2 Partitioning method
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The mixture was shake for 2-3 minutes and left until they separated into two layers. The higher density was located at the bottom of the separating funnel while the lower density was at the top. The solvent mixtures was collected and the aqueous residue was remained. The partitioning extract was repeat for another 2 times (Figure 1). The same method was used for other solvents dichloromethane (DCM), chloroform (CHCl3) and n-Butanol (nBuOH) and distilled water. These solvents and distilled water left were evaporated using the Freeze Dryer (GENEVAC LTO, EZ 2.3 ELITE). Each crude sample obtained from different solvents were weighed and tabulated in the table. The products were stored in refrigerator at -20°C until

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