Abstract Background The purpose of the present study was to enhance the memory-boosting activity of the standardized hydroalcoholic Camellia sinensis extract (CSE) by the formation of nanophytosomes with Leciva S70 phospholipid. The central composite design was used to optimize the solvent evaporation method for the formulation of C. sinesis phytosomes (CSP). Results The optimized formulation had a mean particle size of 212.3 nm ± 0.39, PDI of 0.238 ± 0.0197, and zeta potential of −42.02 ± 0.995 mV. C. sinensis phytosome formation was confirmed by analytical techniques. The aqueous solubility of the developed CSP was 95.92 ± 0.31, which is 7.34 times greater than that of pure CSE (13.07 ± 0.19). CSP was found more effective than either pure CSE (26.42 ± 0.4654%) or the physical mixture (32.15 ± 0.4596%) in releasing the CSE from the formulation (72.16 ± 0.5248%). Acute toxicity study corroborated the safety of CSP in rats. CSP demonstrated a significant (p < 0.05) reduction in escape and transferred latency on both days (15th and 16th) as compared to CSE, indicating the improvement of the memory-boosting activity. Furthermore, CSP-treated rats significantly improved acetylcholine (Ach) levels and brain tissue concentration compared with CSE. Moreover, the phytosomal formulation of CSP exhibited its rationality with an improvement of bioavailability by 3.21 folds compared with pure CSE. Conclusion The presence of phospholipids in the CSP formulation and the formation of smaller particles may aid in crossing the blood–brain barrier, increasing brain tissue concentration and bioavailability. This, in turn, leads to an increase in memory-boosting activity. Graphical abstract

Purpose. The present study aimed to improve the aqueous solubility, permeability, bioavailability, and nootropic potential of standardized Emblica officinalis extract (EOE) by developing a novel phytosomal formulation. Method. Emblica officinalis extract-loaded phytosomes (EOPs) were prepared using solvent evaporation. The EOP was prepared at different molar ratios of extract and phospholipid. Herein, the effects of phospholipid extract ratio (A), temperature (B), and reaction time (C) were systematically investigated on entrapment efficiency using Box-Behnken design. In vitro and in vivo characterizations of the optimized formulation were performed. Results. Optimized EOP formulation (89.90 ± 0.24 μg/ml) exhibited improved aqueous solubility than plain EOE (11.85 ± 0.25 μg/ml). The optimized formulation’s particle size and Zeta potential were 198.4 ± 0.20 nm and −39.0 ± 0.40 mv. DSC and XRD studies confirmed the partial amorphization of EOE in phytosomes. Optimized formulation exhibited 69.82 ± 0.17% of EOE release at 12 h and followed zero-order release kinetics. Moreover, the phytosomal formulation of EOE exhibited its rationality with an improvement of bioavailability by 2.7 folds compared with pure EOE. Compared to EOE, EOP showed significantly (p<0.05 lower escape and transfer latencies on both days in MWMT and EPMT, indicating more effective memory-enhancing activity. Furthermore, EOP-treated rats exhibited improved acetylcholine (Ach) levels than EOE. Brain tissue concentrations measured following EOP oral administration (1.06 ± 0.04 μg/ml) were substantially greater (p<0.05) than those following EOE (0.32 ± 0.07 μg/ml). The brain dopamine and serotonin concentration were found to be higher (16.27 ± 1.209 and 43.28 ± 1.550 ng/ml) in the EOP-treated group as compared to the pure extract-treated group (10.40 ± 1.185 and 32.79 ± 1.738 ng/ml). Conclusion. Improvement of aqueous solubility, permeability, dissolution, bioavailability, and narrower particle size distribution could facilitate enhancement in the nootropic potential of EOE phytosomal formulation.

The goal of this study was to provide a new, easy, accurate, cost-effective, exact, sensitive, specific, robust, and rugged method for quantifying Epigallocatechin 3 gallate in wistar rat plasma using reverse phase high-performance chromatography (RP-HPLC). The stationary phase was a Zorbax SB C18 5 mu (4.6*150) mm column, while the mobile phase was water with 0.1 percent formic acid (A) and acetonitrile (ACN) with 0.08 percent formic acid (B). The experiment was conducted at 30 degrees C with a flow rate of 1.0 ml/min with PDA detectors at 274 nm. With an r2 of 0.9999, the method was shown to be linear in the concentration range of 0.2-25 mu g/ml. At the same retention time (Rt) of epigallocatechin 3 gallate, no interference of co-eluting peaks of endogenous chemicals from the biological matrix was found. The intraday and interday precision RSD (%) was found to be within acceptable limits (less than 2%). The overall mean recovery percentage was determined to be 96.92 %. The LOD and LOQ were determined to be 0.0682 +/- 0.0011 mu g/ml and 0.205 +/- 0.004 mu g/ml, respectively. In short-term and long-term stability tests, auto sampler, bench-top, and freeze-thaw stability tests were found to be stable. The developed approach reported was determined to be well within acceptable limits. As a result, in the future, this method can be successfully employed in clinical laboratories to estimate epigallocatechin 3 gallate alone or in conjunction with other analytes or markers in pharmacokinetic, bioequivalence, and therapeutic drug monitoring.

Objective: Present study aimed to develop and validate a novel, unique, simple, quick, cost-effective, sensitive, specific, accurate, precise, rugged, and robust bioanalytical method for the quantification of gallic acid in rat plasma by reverse phase high-performance liquid chromatography (RP-HPLC) using gradient elution technique. Methods: The stationary phase was a Zorbax SB C18 5 µ (4.6*150) mm column, with the mobile phase being water with 0.1 percent formic acid (A): acetonitrile (ACN) with 0.08 percent formic acid (B). Gradient chromatographic method was used throughout this experiment from the point of view of the estimation of gallic acid from herbal formulations when present along with other phytoconstituents. So at the gradient method, all the present phytoconstituents has cleared off from the column and no any strongly adsorption of phytoconstituents occurred. The experiment was carried out at a flow rate of 1.0 ml/min at 30 °C utilising PDA detectors at 271 nm. The proposed method was validated for different parameters. Results: The approach was found to be linear in the concentration range of 0.5-100 µg/ml, with a r2 of 0.9998. There was not observed any interference of co-eluting peaks of endogenous compounds from the biological matrix at the same retention time (Rt) of gallic acid. The RSD (%) of intra and interday precision was found to be within acceptable limit. The overall % mean recovery was found to be 99.97%. LOD and LOQ were found to be 0.1 and 0.5 μg/ml, respectively. In terms of fluctuation in essential parameters and operating settings, the devised bioanalytical approach was shown to be rugged and resilient. Short-term, long-term, autosampler, bench-top, and freeze-thaw stability experiments revealed that gallic acid is stable. Conclusion: The developed method described in this report was found to be well within an acceptable range. Hence, in the future, this method can be used successfully for the estimation of gallic acid alone or in combination with another analyte or marker present in bulk or an extract containing various phytoconstituents in pharmacokinetic, bioequivalence, and therapeutic drug monitoring studies in clinical laboratories.

Acute toxicity study was carried out for evaluation of toxic effect of formulation, safe acute dose for human and potential target organ for toxicity. Tea (Camellia sinensis), a cultivated evergreen plant and belonging to Theaceae family. The goal of present investigation was to perform acute toxicity study of Camellia sinensis phytosome formulation (CSP) for estimation of LD 50 or Median fatal dose and safe acute dose for human. A total of 12 female wistar rats weighing 90 to 100 gm were used in this investigation and divided into four groups, each with three animals, in accordance with 423 OECD guidelines. Group 1, 2, 3 and 4 animals were orally administered with 300 mg/kg bw of CSP, 300 mg/kg bw of CSP, 2000 mg/kg bw of CSP and 2000 mg/kg bw of CSP respectively. All the animals were observed for clinical sign, symptoms, body weight changes and mortality for 14 days. There was no death in any of the animals, and there were no significant variations in clinical signs or body weight in the experimental group. There were no abnormal alterations found during a gross necropsy.

Emblica officinalis, commonly known as amla, belongs to the Euphorbiaceae family. The herbal medicine phytosome was used to create a novel drug delivery system to extract E. officinalis fruit. E. officinalis fruit extract phytosome (EOP) formulation was tested for toxicity, and the LD50 cut-off value was determined using female wistar rats weighing 95 to 105 g. Acute toxicity testing was performed on a total of 12 female wistar rats, with three rats in each of the four groups as per 423 organisation for economic co-operation and development (OECD) guidelines. A single oral dose of the EOP formulation at 300, 300, 2000, and 2000 mg/kg body weight was administered to groups I, II, III, and IV. For 14 days, we monitored all of the animals to see if any of them died, developed any clinical symptoms, or put on any significant weight. There were no fatalities among the animals, and notable distinctions between the two groups were not found in terms of observed clinical symptoms or animal body weight. At the time of the gross necropsy, no abnormalities were discovered. The phytosome (EOP) formulation of E. officinalis has been shown to be non-lethal and tolerable at 2000 mg/kg bw. 5000 mg (mg/kg) of LD50 were selected for this investigation.

A versatile, accurate, precise and economic method for simultaneous determination of Domperidone and Omeprazole in fixed dose combination products was developed. The absorbance values at 284.8 nm and 301.2 nm and 290.9nm (isoabsorptive point) were used for the estimation of Domperidone and Omeprazole, respectively without mutual interference. This method obeyed Beer's law in the concentration range of 18–28 μg/ml for Domperidone and 5–30 μg/ml for Omeprazole. The results of analyses have been validated statistically for linearity, accuracy and precision, LOD and LOQ of the proposed method.

versatile, accurate, precise and economic method for simultaneous determination of simvastatin and ezetimibe in fixed dose combination products was developed. The absorbance values at 238.2 nm and 247.6 nm and 243.3nm (isoabsorptive point) were used for the estimation of simvastatin and ezetimibe, respectively without mutual interference. This method obeyed Beer's law in the concentration range of 3-18 μg /ml for simvastatin and 5-30 μg /ml for ezetimibe. The results of analyses have been validated statistically for linearity, accuracy and precision, LOD and LOQ of the proposed method. Keywords: Simvastatin (SMV), Ezetimibe (EZE), methanol, distilled water, Simultaneous equation method, Absorption ratio method.

The present work describes a simple Reverse Phase HPLC method for the determination of Omeprazole and Domperidone from capsule formulations. The determination was carried out on a Gasco, ODS, C-18 (250× 4.5 mm, 5 micron) column using a mobile phase of Acetonitrile: 0.05M ammonium acetate buffer (pH- 4) in the ratio of (85:15). The flow rate and runtime were 1 ml/min and 10 min, respectively. The eluent was monitored at 280 nm. The method was reproducible, with good resolution between Omeprazole and Domperidone. The detector response was found to be linear in the concentration range of 4-12 µg/ml for Omeprazole and 8-16 µg/ml for Domperidone. Keyword: Domperidone (DOM), Omeprazole (OME), Acetonitrile, Jasco HPLC (2080 Plus) and 0.05 ammonium acetate buffer (pH - 4)