Study of Gynostema pentaphyllum (Thunb.) Makino Planted from Tissue Culture: Quality Control and Its In Vitro Activity on Human Lymphocytes

Abstract

Plantlets of Gynostema pentaphyllum (Thunb.) Makino from tissue cultures aged 1-2 months grown in a
nursery were transferred to Rai Khrang Dong, Kao Ko, in Petchaboon Province. Soil and water samples from
the selected field for the plantlets to grow were analyzed by the Department of Land Development, Ministry of
Agriculture and Cooperatives. The quality of the land was improved based on the analytical results. The
fresh upper parts of the plantlets were collected twice for analysis of total saponins, i.e., at 3 months old before
flowering and at 4.5 months old, which was the harvesting period when the plants flowered but bore no fruits.
The harvested fresh plants weighed 331 kg. After washing and drying in the oven at about 55°C , the dried
plants weighed 38.4 kg, giving a percentage yield of 11.6. The output was about 1,334 kg/rai of fresh plants, or
153.6 kg/rai of dried plants. The total saponins of the 3 and 4.5-month-old plants were 7.11 percent and 9.00
percent, respectively, while the standard specification of this plant states that total saponins in the crude drug
should be not less than 8 percent. Ginsenoside Rb1 was detected by thin layer chromatography (TLC) as
compared with standard Ginsenoside Rb1. Analysis of powdered crude drug showed no insecticide contamination,
while lead, cadmium and arsenic levels were within limits. Powered crude drug did not pass the test for
microbial contamination, while the water extract met the requirements. In vitro study in human peripheral
blood mononuclear cells showed that Gynostemma pentaphyllum aqueous extract at the concentrations of 1, 5,
10 and 100 μg/ml could stimulate lymphocyte proliferation.