Sheeba Fareed

Current approaches toward production of secondary plant metabolites

Plants are the tremendous source for the discovery of new products with medicinal importance in drug development. Today several distinct chemicals derived from plants are important drugs, which are currently used in one or more countries in the world. Secondary metabolites are economically important as drugs, flavor and fragrances, dye and pigments, pesticides, and food additives. Many of the drugs sold today are simple synthetic modifications or copies of the naturally obtained substances. The evolving commercial importance of secondary metabolites has in recent years resulted in a great interest in secondary metabolism, particularly in the possibility of altering the production of bioactive plant metabolites by means of tissue culture technology. Plant cell and tissue culture technologies can be established routinely under sterile conditions from explants, such as plant leaves, stems, roots, and meristems for both the ways for multiplication and extraction of secondary metabolites. In vitro production of secondary metabolite in plant cell suspension cultures has been reported from various medicinal plants, and bioreactors are the key step for their commercial production. Based on this lime light, the present review is aimed to cover phytotherapeutic application and recent advancement for the production of some important plant pharmaceuticals.

Acute and subacute oral toxicity evaluation of Tephrosia purpurea extract in rodents

Abstract Objective To evaluate the acute and subacute toxicity of 50% ethanolic extract of Tephrosia purpurea (T. purpurea) in rodents. Methods The acute toxicity test was conducted in Swiss albino mice. The extract of T. purpurea was administrated in single doses of 50, 300 and 2000 mg/kg and observed for behavioral changes and mortality, if any. In subacute toxicity study, Wistar rats of either sex were administered two doses of T. purpurea i.e., 200 and 400 mg/kg (One-tenth and one-fifth of the maximum tolerated dose), p.o. for 4 weeks. During 28 days of treatment, rats were observed weekly for any change in their body weight, food and water intake. At the end of 28 days, rats were sacrificed for hematological, biochemical and histopathology study. Results In the acute toxicity study, T. purpurea was found to be well tolerated upto 2000 mg/kg, produced neither mortality nor changes in behavior in mice. In subacute toxicity study, T. purpurea at dose level of 200 and 400 mg/kg did not produce any significant difference in their body weight, food and water intake when compared to vehicle treated rats. It also showed no significant alteration in hematological and biochemical parameters in experimental groups of rats apart from a decrease in aspartate transaminase, alanine transaminase and alkaline phosphate content at the dose of 400 mg/kg. Histopathological study revealed normal architecture of kidney and liver of T. purpurea treated rats. Conclusions These results demonstrated that there is a wide margin of safety for the therapeutic use of T. purpurea and further corroborated the traditional use of this extract as an anti hepatocarcinogenic agent.

Hyphenated chromatographic analysis of bioactive gallic acid and quercetin in Hygrophila auriculata (K. Schum) Heine growing wildly in marshy places in India by validated HPTLC method

Abstract Objective A simple, accurate, and rapid high-performance thin-layer chromatographic (HPTLC) method for simultaneous quantification of the two biologically active flavonoidal compounds, gallic acid and quercetin, in Hygrophila auriculata (K. Schum) Heine (HA) has been established and validated. Methods Chromatography was performed on aluminium foil-backed silica gel 60 F254 HPTLC plates with the binary mobile phase toluene: ethyl acetate: formic acid (5:4:1, v/v/v). Ultraviolet detection was performed densitometrically at the maximum absorbance wavelength, 270nm. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with ICH guidelines. Results The system was found to give compact spots for gallic acid (GA) and quercetin (QE) (Rf value of 0.31 and 0.50, respectively). The limit of detection (23 and 41 ng band−1) limit of quantification (69 and 123 ng band−1), recovery (99.4–99.9 and 98.7–99.4%), and precision (i.e ≤1.98 and 1.97) were satisfactory for gallic acid and quercetin respectively. Linearity range for GA and QE were 100–1000 (r2= 0.9991) and 150–900 ng band−1 (r2= 0.9956) and the contents estimated as 0.28±0.01% and 0.41±0.01% w/w respectively. Conclusions This simple, precise and accurate method gave good resolution from other constituents present in the extract. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing Hygrophila auriculata (K. Schum) Heine.

Simultaneous HPTLC-UV530 nm analysis and validation of bioactive lupeol and stigmasterol in Hygrophila auriculata (K. Schum) Heine

Abstract Objective To analyse the two marker compounds lupeol (LP) and stigmasterol (ST) from methanolic extract of Hygrophila auriculata (H. auriculata). Methods Separation was achieved on aluminium plates precoated with silica gel 60F254 with toluene-methanol-formic acid (7.0: 2.7: 0.3 v/v/v) as mobile phase. Results Densitometric analysis was performed at 530 nm in the reflectance mode. Compact bands for LP and ST were obtained at RF 0.52 ± 0.02 and 0.28 ± 0.05. Linearity (r2=0.998 5 and 0.993 7), limit of detection (45 and 18 ng/band) limit of quantification (135 and 54 ng/band), recovery (98.2%–99.7% and 97.2.2%–99.6%), and precision ( Conclusion The method demonstrated efficient analysis testing of LP and ST in samples; therefore it can be used for routine analysis.

Nanoemulsion: for improved oral delivery of repaglinide

Abstract Repaglinide (RPG) is a fast-acting prandial glucose regulator. It acts by stimulating insulin release from pancreatic β-cells. Recurrent dosing of RPG before each meal is burdensome remedy. Hence the plan of the present study was to evaluate nanoemulsion as a hopeful carrier for RPG for persistent hypoglycemic effect. The drug was incorporated into oil phase of nanoemulsion to give improved biopharmaceutical properties as compared to the lipid-based systems. Pseudo ternary phase diagrams were prepared by aqueous titration method. Formulations were selected at a difference of 5% w/w of oil from the o/w nanoemulsion region of phase diagrams. The optimized nanoemulsion formulation constituted sefsol-218 (5% v/v) as an oil phase, 30% v/v of Tween-80 and transcutol as a surfactant and co-surfactant to restrain nanodroplet size and low viscosity and distilled water (65%). In vitro dissolution studies showed higher drug release (98.22%), finest droplet size (76.23 nm), slightest polydispersity value (0.183), least viscosity (21.45 cps) and immeasurable dilution capability from the nanoemulsion as compared with existing oral tablet formulation. The optimized RPG nanoemulsion formulation showed better hypoglycemic effect in comparison to tablet formulation in experimental diabetic rats. No significant variations were also observed in the optimized formulation when subjected to accelerated stability study at different temperature and relative humidity over a period of 3 months.

Validation of the method for the simultaneous estimation of bioactive marker gallic acid and quercetin in Abutilon indicum by HPTLC

Abstract Objective To establish and validate an simultaneous estimation of the two biomarker compounds gallic acid (GA) and quercetin (QE) from methanolic extract of Abutilon indicum (AI). Methods Chromatography was performed on aluminium foil-backed silica gel 60 F254 HPTLC plates with the binary mobile phase toluene-ethyl acetate-formic acid (5:4:1, v/v/v). Ultraviolet detection was performed densitometrically at the maximum absorbance wavelength, 270nm. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with ICH guidelines. Results The system was found to give compact spots for GA and QE (Rf value of 0.31 and 0.50, respectively). The limit of detection (23 and 41 ng band-1) limit of quantification (69 and 123 ng band-1), recovery (99.4–99.9 and 98.7–99.4%), and precision (≤1.98 and 1.97) were satisfactory for gallic acid and quercetin respectively. Linearity range for GA and QE were 100-1000 (r 2 = 0.9991) and 150–900 ng band-1 (r 2 = 0.9956) and the contents estimated as 0.69% ± 0.01% and 0.57% ± 0.01% w/w respectively. Conclusions This simple, precise and accurate method gave good resolution from other constituents present in the extract. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing AI.

Chemopreventive effect of Fumaria indica that modulates the oxidant-antioxidant imbalance during N-nitrosodiethylamine and CC14-induced hepatocarcinogenesis in Wistar rats

Abstract Objective To investigation the chemopreventive efficiency of Fumaria indica extract (FIE) on the antioxidant status of N-nitrosodiethylamine (NDEA) and CCl 4 -induced hepatocarcinogenesis in Wistar rats. Methods The experimental animals were divided into six groups ( n = 6). HCC was induced by single intraperitoneal injection of NDEA in normal saline at a dose of 200 mg/kg body weight followed by weekly subcutaneous injections of CCl 4 (3 mL/kg/week) for 6 weeks, as the promoter of carcinogenic effect. After administration of the carcinogen, 200 and 400 mg/kg of FIE were administered orally once a day throughout the study. At the end of 20 weeks, the body weight, liver weight and relative liver weight were measured. The level of hepatic malondialdehyde (MDA) formation, reduced glutathione (GSH) and the activities of antioxidant enzymes viz. CAT, SOD, GPx, and GST in the liver of NDEA and CCL 4 -treated rats were assessed. Results Obtained results demonstrated that the cotreatment with FIE (200 and 400 mg/kg) significantly prevented the decrease of the body weight and also increased in relative liver weight caused by NDEA and CCl4 administration. FIE also significantly prevented hepatic malondialdehyde (MDA) formation and reduced glutathione (GSH) in NDEA-treated rats which were dose dependent. Additionally, FIE also markedly increased the activities of antioxidant enzymes such as CAT, SOD, GPx, and GST in the liver of NDEA and CCL 4 -treated rats. Conclusions These finding powerfully supports that Fumaria indica exert a chemopreventive effect by reversing the oxidant-antioxidant imbalance during hepatocarcinogenesis induced by NDEA and CCl 4 .

Evaluation of chemopreventive effect of Fumaria indica against N-nitrosodiethylamine and CCl4-induced hepatocellular carcinoma in Wistar rats

OBJECTIVE To investigation the chemopreventive potential of Fumaria indica (F. indica) extract (FIE) on N-nitrosodiethylamine and CCl(4)-induced hepatocarcinogenesis in Wistar rats. METHODS The experimental animals were divided into six groups (n=6). Hepatocellular carcinoma was induced by single intraperitoneal injection of N-nitrosodiethylamine (NDEA) in normal saline at a dose of 200 mg/kg body weight followed by weekly subcutaneous injections of CCl(4)(3 mL/kg/week) for 6 weeks, as the promoter of carcinogenic effect. After administration of the carcinogen, 200 and 400 mg/kg of FIE were administered orally once a day throughout the study. At the end of 20 weeks, the body weight, liver weight and relative liver weight were measured. The percentage of nodule incidence and liver cancer markers such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), γ-glutamyl transferase (γ-GT), total bilirubin level (TBL), α-feto protein (AFP) and carcinoembryonic antigen were estimated along with histopathological investigation in experimental groups of rats. RESULTS Obtained results demonstrated that the cotreatment with FIE significantly prevented the decrease of the body weight and also increased in relative liver weight caused by NDEA. The treatment with FIE significantly reduced the nodule incidence and nodule multiplicity in the rats after NDEA administration. The levels of liver cancer markers such as AST, ALT, ALP, γ-glutamyl transferase, TBL, AFP and carcinoembryonic antigen were substantially increased by NDEA treatment. However, FIE treatment significantly reduced the liver injury and restored the entire liver cancer markers. Histological observations of liver tissues too correlated with the biochemical observations. CONCLUSIONS These finding powerfully supports that F. indica exert chemopreventive effect by suppressing the tumor burden and restoring the activities of hepatic cancer marker enzymes on NDEA and CCl(4)-induced hepatocarcinogenesis in Wistar rats.

Chemopreventive evaluation of Tephrosia purpurea against N-nitrosodiethylamine-induced hepatocarcinogenesis in Wistar rats

Objectives  The chemopreventive potential of Tephrosia purpurea extract (TPE) on N‐nitrosodiethylamine (NDEA)‐induced hepatocellular carcinoma (HCC) in Wistar rats was assessed.

Nanomulsion as a Carrier for Efficient Delivery of Metformin

Metformin (MTF) improves hyperglycemia primarily by suppressing glucose production by the liver. The objective of our investigation was to evaluate nanoemulsion as a promising carrier for MTF for sustained hypoglycemic effect. The drug was incorporated into oil phase of nanoemulsion, which finally improved biopharmaceutical properties achieved when compared with lipid based systems. Pseudo ternary phase diagrams were prepared by aqueous titration method. Formulations were selected at a difference of 5% v/v of oil from the o/w nanoemulsion region of phase diagrams, and then thermodynamic stability and dispersibility tests were performed. The composition of optimized formulation was hydrogenated castor oil (5% v/v), 30% v/v of surfactant (tween 80), co-surfactant (transcutol) and distilled water (65% v/v) as an aqueous phase. The preparation showed maximum drug release (98.70%), optimal globule size (92.25 nm), lowest polydispersity value (0.172), lesser viscosity (22.124 cps) and infinite dilution capability. The antidiabetic activity of optimized MTF nanoemulsion formulation evaluated by blood glucose estimation showed significant hypoglycemic effect which was comparable to that observed with conventional marketed formulation in experimental diabetic rats. Optimized formulation was subjected to stability studies at different temperature and relative humidity and was found to be stable. No significant variations were observed in the formulation over a period of 3 months at accelerated storage conditions.