Page 115 - วารสารการแพทย์แผนไทย ปีที่ 21 ฉบับที่ 2

J Thai Trad Alt Med Vol. 21 No. 2 May-Aug 2023 331

1.2 Virus with distilled water two times consecutively.
SARS-CoV-2 virus (Delta variant/EPI_ The aqueous extract was then pooled, concen-

ISL_3797061) was obtained from a human na- trated and dried by rotary evaporation yielding
sopharyngeal swab. The virus was propagated PSP, Y5R and CTL extracts. The dried extracts
in Vero E6 cells by three passages to establish were collected and kept in a light-protected

a high-titer stock and stored at -80˚C for use bottle at -20˚C until used.
in all experiments. Virus titration as TCID50 The extracts were also analyzed for their
titer/mL was performed. All experiments with phytochemical compounds for quality control

live SARS-CoV-2 virus were performed at a using Thin layer chromatography (TLC) using
certified biosafety level 3 facility, the National reverse phase, RP-18 plates (Merck KGaA,
Institute of Health, Department of Medical Darmstadt, Germany) as the stationary phase

Sciences, Thailand. and the mixture of n-hexane: acetone: ethyl
1.3 Standards and Drug formulations acetate (70:20:5 v/v) as the mobile phase. The

B-eudesmol (> 98% HPLC grade) was drugs were monitored at 254 nm (A), 366 nm
form Merck KGaA (Darmstadt, Germany). Ya (B) and were sprayed with anisaldehyde sul-
Prasa Pro Yai and Ya Ha Rak formulations phuric acid reagent. β-eudesmol was used as

were kindly gifted by Mr. Kaisee Limprasert, a standard. The plant extracts and the quality
Thai Traditional Medicine (พทว.13150 พท.ภ. control data were conducted at the Herbal

13761 พท.ผ. 2001 บป.20). Herbal formulations Quality Assurance Center, Medicinal Plant
were produced using the traditional household Research Institute, Department of Medical
medicine formulation (2018) as listed in the Sciences.

National List of Essential Medicines. (Herbal 2.2 MTT assay
3
medicine list, 2012); lot July 2021. The Ya The cells were seeded at 5 5 10 cells in
Chanthalila formulation was purchased from 100 µL medium per well of 96–well plates and

Vejpong Pharmacy (HOCK AN TANG) com- left in the incubator for 24 hours. The medium
pany limited (Thailand); lot July 2021. was removed and the cells were treated with
eight different concentrations of each com-
2. Methods pound in triplicate and then incubated for 48

2.1 Extract production hours. The medium was replaced with 200 µL

Total 300 g of powdered plant was re- MTT reagent and then continued to incubate
fluxed with distilled water. The resultant solu- for 4 hours. The MTT solution was discarded
tions were then filtered and further extracted and 200 µL DMSO was added to each well to