Page 128 - วารสารกรมการแพทย์แผนไทยฯ ปีที่ 20 ฉบับที่ 1

108 วารสารการแพทย์แผนไทยและการแพทย์ ทางเลือก ปีที่ 20 ฉบับที่ 1 มกราคม-เมษายน 2565

G6PD enzyme-deficient human erythrocytes pended in DMSO to prepare the stock solution.
against oxidative hemolysis. Based on our 2.2 total phenolic content
knowledge, this is the first report about the The total phenolic content was deter-

assessment of antileukemic activity, inhibition mined by using the Folin-Ciocalteu assay .
[16]
of Heinz body formation, hemolytic effect, and The crude extract was prepared to yield a

protective activity of this extract. concentration of 1 mg/mL. About 100 μL of the
extract (1 mg/mL) was combined and mixed
Methodology with 0.75 mL of the Folin–Ciocalteu reagent in

1. Materials the test tube. The liquid mixture was allowed

1.1 plant material to stand for 5 minutes at room temperature.

L. leucocephala young leave (3-4 layers The mixture was then added about 0.75 mL
from shoot) were collected from Western Uni- of sodium carbonate (Na CO ), and the test
3
2
versity, Kanchanaburi, Thailand in December tube was shaken gently to mix them. After
and authenticated by Manop Poopath (Senior 90 minutes, the absorbance against the re-
Botanists, Forest and Plant Conservation Re- agent blank was measured at 725 nm with the
search Office, Department of National Parks, UV-Vis spectrophotometer.

Wildlife and Plant Conservation, Thailand). Gallic acid (range of concentration from
0.1 to 10 mg/mL) was used to calibrate the
2. Methods standard curve. Total phenolic content was re-

2.1 preparation of extracts vealed as milligrams of gallic acid equivalents
The fresh young leaves were dried in a per grams of the extract sample (mg GAE/g

hot air oven at 60˚C. The dried leaves were extract).
grounded and extracted by the maceration 2.3 blood samples collection
method. The powder was soaked using The human test procedure was approved

80% ethanol at 1:10 w/v. The mixture was by the Western University Ethics Committee
macerated for four days. The extract was on Human Research (WTU 2564-007).

filtered. The filtrate was evaporated using a Human peripheral blood (3 mL) from 17
rotary evaporator at 70˚C, thus leaf condensed G6PD deficient (6 male and 11 female) and 20
extract was obtained. The condensed extract healthy donors (10 male and 10 female) was

was heated in a water bath to obtain an extract collected after their consent. Erythrocytes
with a solid. Finally, the crude extract was kept were separated by centrifugation and then
in the refrigerator at -20˚C until used and sus- suspended in sterile PBS to obtain the 10%