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...