Sarfaraz Ahmed

High-performance thin-layer chromatography based concurrent estimation of biomarkers ent-phyllanthidine and rutin in the dried aerial parts of Flueggea virosa

The biomarkers are needed to be defined for standardization purposes so that safe and effective herbal formulations can be catered to the society. There is an urgent need for statistical support of herbal drugs because most of the herbal products are still used in the non-standardized form. This study is based on the development of a simple and sensitive RP-HPTLC method for concurrent estimation of two biomarkers ent-phyllanthidine and rutin in the methanol extract of aerial parts of Flueggea virosa. The developed method was found to be simple, economic and sensitive. Separation and quantification were performed with acetonitrile: water (4:6 V/V) used as the mobile phase on glass-backed RP-HPTLC plate. Detection of absorption maxima and quantification was done at 310 nm of UV region. The developed chromatographic system was found to give a sharp band for ent-phyllanthidine and rutin at Rf 0.73 ± 0.01 and 0.68 ± 0.01, respectively. The linearity ranges for ent-phyllanthidine, and rutin were found to be 200–1600 ngband−1 and 100–1400 ngband−1, respectively, with correlation coefficients (r2 values) of 0.998 and 0.997, respectively. The percentage of ent-phyllanthidine and rutin was found to be 9.121 ± 0.02% and 1.018 ± 0.04% (w/w), respectively. The resolution of bands and separation of constituents in FVME exhibited the perfect optimization of the developed method. The validation statistics supports the proposed method for standardizing crude drugs as well as formulations of a natural product containing ent-phyllanthidine and rutin.

Neoclerodane Diterpenoids from Reehal Fatima, Teucrium yemense

Teucrium yemense (Defl), locally known as Reehal Fatima, is a medicinal plant commonly grown in Saudi Arabia and Yemen. Phytochemical investigation of the aerial parts of T. yemense yielded six new neoclerodane diterpenoids, namely fatimanol A-E (1, 2, 3, 5, and 6) and fatimanone (4), and the known teulepicephin (7). As both the Teucrium genus and the related Lamiaceae family have previously been widely reported to possess anthelmintic and antimicrobial activities, the structural and biological characterization of the seven diterpenoids was pursued. The structures of the new compounds were elucidated from their 2D NMR and MS profiles and by comparison to related compounds. The structure of fatimanol D (5) was confirmed by X-ray crystallographic analysis. The new structures contribute to the breadth of knowledge of secondary metabolites in this genus.

Studies on wound healing activity of some Euphorbia species on experimental rats

Background: Plants of Euphorbiaceae are used in folkloric medicines in variety of ailments and well known for chemical diversity of their isoprenoid constituents. This study was carried out to explore the preliminary wound healing potential of four Euphorbia species (E. consorbina 1, E. consorbina 2, E. inarticulata, E. balsamifera and E. schimperi). Materials and Methods: Excision wound surface of the animals were topically treated with ethyl acetate and methanol extracts of plants at a dose of 400 mg/kg body weight for twenty days. Povidone-iodine ointment was used as a reference drug. Wound contraction measurement and period of epithelialization were used to assess the effect of plants extracts on wound repairing. Results: The groups treated with methanol extracts of E. balsamifera and E. schimperi showed profound effects, high rate of wound contraction (100%) and decrease in epithelization period 19.00±0.40 and 18.50±0.64 respectively, followed by methanol extracts of E. consorbina 2, ethyl acetate extract of E. inarticulata and ethyl acetate extracts of E. consorbina 2 which showed significant (P <0.001) wound contraction and decrease in epithelization period. Conversely ethyl acetate extract of E. consorbina 1, E. balsamifera and E. schimperi and methanol extract of E. Consorbina 1 and E. Inarticulata treated groups was not showing significant wound healing. Methanol extracts of E. balsamifera and E. schimperi were also tested for their safety margin and found safe up to dose of 2000mg/kg body weight. Conclusion: Topical application of methanol extracts of E. balsamifera and E. schimperi have potential wound healing activity which is identical with standard drug Povidone-iodine.

Simultaneous quantification of two bioactive flavonoids, homoeriodictyol and persicogenin, in the methanol extract of the aerial parts of two different species of genus Rhus by a validated high-performance thin-layer chromatographic-densitometric method

A simple and sensitive high-performance thin-layer chromatographic (HPTLC)—densitometric method was developed for the simultaneous quantification of two flavonoid compounds, persicogenin and homoeriodictyol, in the methanol extracts of the aerial parts of two species (Rhus retinorrhoea and Rhus tripartita) of the genus Rhus grown in the Kingdom of Saudi Arabia. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates using toluene-ethyl acetate-methanol (8:2:0.5, v/v) as the mobile phase. Scanning and quantification were done at 293 nm. The system was found to give compact spot for homoeriodictyol and persicogenin at, RF = 0.30 ± 0.01 and 0.48 ± 0.01, respectively. The linearity ranges for homoeriodictyol and persicogenin were found to be the same (100–800 ng spot−1) with correlation coefficients (r2 values) of 0.9989 and 0.9983, respectively. The limit of detection (LOD) for homoeriodictyol and persicogenin was found to be 26 and 31 ng band−1, respectively, while the limit of quantification (LOQ) was found to be 77 and 92 ng band−1, respectively. Homoeriodictyol (7.06%) and persicogenin (2.33%) were only found in R. retinorrhoea. The developed method was found to be accurate and precise; hence, it can be used as an important tool to assure the therapeutic dose of homoeriodictyol and persicogenin in herbal formulations as well as for the standardization and quality control of bulk drugs.

Inter-species comparative antioxidant assay and HPTLC analysis of sakuranetin in the chloroform and ethanol extracts of aerial parts of Rhus retinorrhoea and Rhus tripartita

Abstract Context: Extensive research on Rhus (Anacardiaceae) shows their antioxidant potential, which warrants further evaluation of its other species. Objective: To perform a comparative antioxidant assay on extracts of R. retinorrhoea and R. tripartita, including sakuranetin quantification by a validated HPTLC method. Materials and methods: In vitro antioxidant assay was performed on chloroform and ethanol extracts of R. retinorrhoea Steud. ex Oliv. (RRCE and RREE) and R. tripartita (Ucria) Grande (RTCE and RTEE) by DPPH radical scavenging (at 31.25, 62.5, 125, 250 and 500 μg/mL concentrations) and β-carotene-linoleic acid bleaching methods at 500 μg/mL concentration. Densitometric HPTLC method was developed and validated using toluene: ethyl acetate: methanol (8:2:0.2; v/v/v) as mobile phase, executed on glass-backed silica gel F254 plate and scanned at 292 nm. Results: Antioxidant activity of Rhus extracts tested by the two methods (DPPH/BCB) was found in order of RTEE > RREE > RTCE > RRCE with IC50 118.67/256.26, 315.75/82.35, 827.92/380.0 and 443.69/292.75, respectively. Scanning of the HPTLC plate provided an intense peak of sakuranetin at Rf = 0.59. The estimated sakuranetin content in the dry weight of the extracts was highest in RREE (27.95 μg/mg) followed by RRCE (25.22 μg/mg), RTEE (0.487 μg/mg) and RTCE (0.0 μg/mg). Presence of sakuranetin in RREE, RRCE and RTEE supported the highest antioxidant property of the two Rhus species. Nonetheless, low sakuratenin in R. tripartita indicated the presence of other bioactive constituents responsible for synergistic antioxidant activity. Conclusion: The developed HPTLC method therefore guarantees its application in quality control of commercialized herbal drugs and formulations containing sakuranetin.

Chemical Constituents of Cleome rupicola Growing in Saudi Arabia

Phytochemical investigations of extracts from the aerial parts of Cleome rupicola resulted in the isolation of two new and three known secondary metabolites corresponding to three secodammarane triterpenes: cleomdiolic acid (1), shoreic acid (3), foveolins B (4), and two flavonol glycosides: isorhamnetin-3,7-O-α-L-dirhamnoside-3′′-O-acetyl (2) and isorhamnetin-3-β-D-glucoside-7-α-L-rhamnoside (5). The structures of these compounds were characterized and identified by spectral 1D and 2D NMR and MS analyses in addition to comparison with spectral data of related known compounds. Compounds 1 and 2 are new. This is the first phytochemical investigation of the extract of the aerial part of Cleome rupicola. Bioactivity assay on HeLa cells showed the cytotoxicity of the new compounds (1) (IC50 = 64.7 μg/mL) and (2) (IC50 = 112.6 μg/mL).

Two New Sesquiterpene Alcohols Isolated from Senecio hadiensis Forssk. Grown in Saudi Arabia

Phytochemical investigation on the aerial parts of Senecio hadiensis Forssk. led to the isolation of two new sesquiterpenoids, presilphiperfolan‐2α,5α,8α‐triol (1) and presilphiperfolan‐2α,5α,8α,10α‐tetraol (2) featuring the rare presilphiperfolane‐type frameworks. The structures of 1 and 2 were elucidated on the basis of extensive spectroscopic (1D‐ and 2D‐NMR, HR‐ESI‐MS) methods and by comparison with the literature data. The isolates 1 and 2 were evaluated in‐vitro for antiinflammatory, cytotoxic, and peroxisome proliferator activated receptor alpha and gamma (PPARα and PPARγ) agonistic activities.