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The mixtures of Derris scandens (DZSS) formula is a Thai traditional medicine, which consists of Derris scandens (Roxb.) Benth. (Fam.Leguminosae-Papilioideae) (D), Zingiber cassumunar Roxb. (Fam.Zingiberaceae) (Z), Suregada multiflora Baill. (Fam.Euphorbiaceae) (S) and Siphonodon celastrineus (Fam.Celastraceae) (S). According to the Thai National List of Essential Herbal Medicine (2013), there are two traditional formulas of DZSS formula, DZSS1 and DZSS2, which contains a different amount of Zingiber cassumunar Roxb. rhizome. The ratios of 4 herbs (D:Z:S:S) in the DZSS1 and DZSS2 formulas are 1:1:1:1 and 1:2:1:1, respectively. 1,2 The DZSS formula has been traditionally used for relieving muscle pain. The formula is given orally at the dosage of 900-1,500 mg three times a day immediately after a meal. This medicinal herb formula is contraindicated in pregnant women and should be carefully used in patients with peptic ulcer since D. scandens showed a similar mechanism of anti-inflammation as that of non-steroidal anti-inflammatory drugs (NSAIDs) with an inhibition against prostaglandin production. 1,2 Gastrointestinal side effects are thus the major disadvantage of the oral DZSS formula. It is therefore interesting to develop the DZSS formula in a topical dosage form. Previously literature reviewed on the subject of single ingredients of the DZSS formula indicated the presence of coumarins, isoflavones, flavones, isoflavone glycosides and phenyl coumarins as chemical constituents from D. scandens. 3-5 D. scandens, has been used in Ayurvedic, Thai and Chinese herbal medicine to treat a variety of painrelated conditions, including muscle pain, joint pain, arthritis and headaches. Dalpanitin and vicenin-3, two of the flavonoids isolated from D. scandens gave MICs of 23 μg mL-1 against S. aureus. Dalpanitin also exhibited relevant MICs on Gramnegative bacteria (94 μg mL-1 against Escherichia coli and Pseudomonas aeruginosa). 6 For the Thai traditional medicine, volatile oil from Z. cassumunar has been used directly apply and penetrate on the skin for remedied for muscle stress and joint pain. 7 Four phenylbutanoids including (E)-4(3´,4´dimethoxyphenyl)but-3-en-1-ol (compound D), (E)-(3´,4´-dimethoxyphenyl)but-3-en-1-yl acetate (D-acetate), (E)-1(3,4-dimethoxyphenyl)butadiene (DMPBD) and (E)-3(3´,4´-dimethoxyphenyl)-4-[(E)-3´,4´-dimethoxystyr yl]cyclohex1-en (DMPDMS) that isolated from Z. cassumunar extract. 8,9 The stem of S. multiflora and S. celastrineus have been used for treatment of skin disease. The bark of S. multiflora found chemical constituents such as two ent-kaurene diterpenes, ent-16-kaurene-3β,15β,18-triol and ent-3-oxo-16-kaurene-15β,18-ABSTRACT Background: The mixtures of Derris scandens (DZSS) formula is a Thai traditional medicine, which consists of 4 medicinal plants, including Derris scandens (Roxb.) Benth. (D) Zingiber cassumunar Roxb. (Z), Suregada multiflora Baill. (S) and Siphonodon celastrineus (S). The DZSS formula has been used in an oral dosage form for the treatment of muscle pain. However...

Background and purpose: Benjakul, a traditional Thai formulation for cancer treatment, is composed of five plants. This study aimed to assess the cytotoxicity of Benjakul, its five plants, and its isolated compounds against non-small cell lung cancer (NSCLC) by the sulforhodamine B (SRB) assay. Experimental approach: Analyses of cell cycle and membrane asymmetry changes were performed with different fluorescent dyes and analyzed by flow cytometry in NCI-H226 cells. Activation of caspase-3 was measured using a caspase-3 colorimetric assay kit. The pan-caspase inhibitor Z-VAD-FMK was used in analyses of cell cycle and caspase-3 activity. Findings/Results: Benjakul exhibited cytotoxicity against NSCLC with IC50 between 5.56-5.64 μg/mL. Among its five ingredients, Benjakul displayed the highest selectivity with selectivity index values ranging from 2.93 to 6.88, with the exception of Plumbago indica , indicating its protective effects. Plumbagin and 6- shogaol displayed the highest cytotoxicity and underwent molecular studies in NCI-H226 cells. Flow cytometry analysis revealed that Benjakul and 6-shogaol dose-dependently induced G2/M phase arrest, and plumbagin dose-dependently induced S-G2/M phase arrest with the highest percentage in early incubation time (12-24 h). At the highest doses, Benjakul extract, 6-shogaol, and plumbagin time-dependently increased the population of sub-G1 apoptotic cells with the highest percentage in longer incubation time (60-72 h). Similarly, membrane asymmetry changes showed time-dependent increases in the percentage of early and late apoptotic cells. Moreover, the apoptosis-inducing effect of Benjakul, 6-shogaol, and plumbagin at the highest dose, via the caspase cascade was confirmed by time-dependent induction of caspase-3 activity, followed by its complete reduction and abolished sub-G1 peaks upon addition of Z-VAD-FMK. Conclusion and implication: Our findings demonstrated for the first time the effects of Benjakul and its compounds on S-G2/M or G2/M phase arrest and caspase-dependent apoptosis in lung cancer cells.

A new reversed-phase high-performance liquid chromatographic method with ultraviolet detection (RP-HPLC-UV) for simultaneous determination of phenytoin impurities, benzophenone and benzil, was developed and validated according to the International Council for Harmonization (ICH) guidelines. Chromatographic separation was performed on a C 8 column using acetonitrile-1% acetic acid (60:40, v/v). The correlation coefficients of the calibration lines were greater than 0.999 with 95% confident interval of y-intercept over the origin. The analytical method showed good precision, intra-day precision ≤1.00 and inter-day precision ≤1.53. The standard solution of each compound exhibited good stability 99.18-99.70%, after storage at room temperature for 24 h. The limit of detection (LOD) and limit of quantification (LOQ) were 0.0015 and 0.005 μg/mL, respectively. The resolution of the impurities was 2.935 ± 0.009. The proposed analytical method was successfully applied to determine the amount of benzophenone and benzil in marketed products. The amount of benzophenone was found at 3.09-5.91 × 10 −3 %, while benzil was not detected in the samples.