Background: Scrophularia umbrosa is a plant used as a traditional herb. Objectives: This study was designed to investigate the antimalarial activity and free radical scavenging of methanol, DCM and nHexane extract of aerial parts and seed, as well as total phenol, total flavonoid content, and phytochemical identification. Methods: GC-MS techniques were used for identification of the chemical composition of the DCM and n-Hexane extract as well as HPLC and NMR techniques for methanolic extract. The hemozoin formation has been suggested as a desirable drug target to antimalaria sieve scheme. In the present study, we used the spectrophotometric assay to screen plant extracts. The DPPH method has been used for antioxidant activity screening of the extracts and fractions. AlCl3 and Folin -Ciocalteu reagents, respectively, were used for total flavonoid and total phenol content of the samples. Results: Our results showed that n-hexane and the MeOH extracts, aerial part, and seed did not display any impressive prevention of heme biocrystallization confidants, however, the DCM extract of aerial part, in comparison with the blank, showed slightly antimalarial effects. Furthermore, fractions (20%, 40%, and 60%) of MeOH extracts of aerial part and seed of S. umbrosa had moderate to strong antioxidant activity and it is possible to conclude that there is a positive relation between phenolic and flavonoid composition as well as antioxidant activity. GC-MS data showed that volatile portions of DCM and n-Hexane extracts of various organs from S. umbrosa are containing few identifiable compounds. In addition, Aucubin and Luteolin-7-O-rutinoside were also recognized from MeOH extracts. Conclusions: Based on the results, the fractions of extracts could be evaluated by 1HNMR for predicting of groups of natural compounds as well as interfacing chemical and biological assessments.

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Scrophularia subaphylla ( S. subaphylla ) L., a medicinal plant from the Scrophulariaceae family, has been reported to possess potential profits in the treatment and prophylaxis of different diseases. Some phenolic compounds in this genus have been displayed decent effects on different types of cancer via multiple mechanisms. The current study aimed to bioassay guided isolation of cytotoxic constituents from the aerial parts of S. subaphylla against breast (MCF-7) and colon (HT-29) cancer cell lines as well as normal cells (L929). Different extracts of S. subaphylla were acquired by Soxhlet apparatus and then subjected to brine shrimp lethality test and MTT assay for assessing their cytotoxic characteristics. Cytotoxic extract subjected to further phytochemical fractionation using solid phase extraction, reversed-phase high pressure liquid chromatography (RP-HPLC), and one dimensional nuclear magnetic resonance (1D-NMR) spectroscopy. The biological activity of the isolated pure components, verbascoside and 3‘ O rhamnosyl -4‘ O para coumaryl 7- hydroxyl salidroside , was assessed using MTT assay against MCF-7 and HT-29 carcinoma cells. Two known phenylpropanoid compounds were isolated from this species. Their structures were elucidated by spectroscopic data (using 1 H-NMR and 13 C-NMR) and compared with the previous literature. Both pure compounds in comparison with control group demonstrated significant antiproliferative activity against cancerous cells ( P < 0.001). In our study, verbascoside and its derivative could inhibit proliferation of cancerous cells without any side effects on normal cells.

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