Biodegradation of Alprazolam in Pharmaceutical Wastewater Using Mesoporous Nanoparticles-Adhered Pseudomonas stutzeri

The release of pharmaceutical wastewaters in the environment is of great concern due to the presence of persistent organic pollutants with toxic effects on environment and human health. Treatment of these wastewaters with microorganisms has gained increasing attention, as they can efficiently biodegrade and remove contaminants from the aqueous environments. In this respect, bacterial immobilization with inorganic nanoparticles provides a number of advantages, in terms of ease of processing, increased concentration of the pollutant in proximity of the cell surface, and long-term reusability. In the present study, MCM-41 mesoporous silica nanoparticles (MSN) were immobilized on a selected bacterial strain to remove alprazolam, a persistent pharmaceutical compound, from contaminated water. First, biodegrading microorganisms were collected from pharmaceutical wastewater, and Pseudomonas stutzeri was isolated as a bacterial strain showing high ability to tolerate and consume alprazolam as the only source for carbon and energy. Then, the ability of MSN-adhered Pseudomonas stutzeri bacteria was assessed to biodegrade alprazolam using quantitative HPLC analysis. The results indicated that after 20 days in optimum conditions, MSN-adhered bacterial cells achieved 96% biodegradation efficiency in comparison to the 87% biodegradation ability of Pseudomonas stutzeri freely suspended cells. Kinetic study showed that the degradation process obeys a first order reaction. In addition, the kinetic constants for the MSN-adhered bacteria were higher than those of the bacteria alone.

The Standardized Wood Extract of Albizia myriophylla: Its Potential as an Active Ingredient in an Anti-inflammatory Herbal Gel Formulation

Albizia myriophylla has been used in Thai folk medicine for treating inflammation-related diseases. The wood of this medicinal plant is traditionally used as a single herbal drug in the form of an aqueous decoction and as a component in several Thai herbal formulations for the remedy of fever, sore throat, and aphthous ulcers. This study aimed to evaluate in vivo the anti-inflammatory potential and possible mechanism of action of the standardized wood extract of A. myriophylla as well as to investigate the anti-inflammatory activity and physicochemical properties of the developed herbal gel formulation containing standardized wood extract of A. myriophylla. Results of quantitative HPLC analysis demonstrated that the standardized wood extract of A. myriophylla contained 22.95 mg/g of 8-methoxy-7,3′,4′-trihydroxyflavone, a bioactive marker compound of A. myriophylla. The standardized wood extract of A. myriophylla (1% w/v) exhibited remarkable inhibition (54.4 – 80.3%) in the croton oil model of topical inflammation at all assessment times, comparable to standard indomethacin (55.3 – 63.6%). Real-time quantitative reverse transcription-polymerase chain reaction was performed to clarify the anti-inflammatory mechanism of standardized wood extract of A. myriophylla, and the result showed that this standardized extract decreased lipopolysaccharide-induced nitric oxide synthase mRNA levels in a dose-dependent manner. The developed herbal gel containing standardized wood extract of A. myriophylla (1% w/w) had good physicochemical characteristics and exhibited potent inhibition (51.4 – 77.8%) of inflammation in a rat ear edema model at all assessment times, comparable to indomethacin gel (33.3 – 40.5%). The notable anti-inflammatory activity of standardized wood extract of A. myriophylla and its developed herbal gel formulation indicates their potential application as natural anti-inflammatory agents.

Development of Simultaneous Analysis Method for Multi-Compounds Content of New Shilajit Using HPLC-UV and the Cognitive Enhancing Effect: Mongolian Shilajit

Shilajit has a longstanding use as an anti-aging and memory enhancing drug. It is known to have excellent anti-bacterial effects and is believed to be effective for cognitive enhancement, but is difficult to standardize because of the lack of quality control standards. This study, for the first time, proposes a quality control standard using a simultaneous analytical method for the drug’s multi-compound content using high-performance liquid chromatography-ultraviolet detection (HPLC-UV) as an aid for the internationalization of Mongolian Shilajit. Phenolic compounds 1-6 were isolated from Mongolian Shilajit extract using bioassay-guided isolation, and the isolated compounds were evaluated for cognitive-related anti-Alzheimer’s disease (AD) activities using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), β-site amyloid precursor protein-cleaving enzyme 1 (BACE1), and advanced glycation end-product (AGE) formation assays. The isolated compounds showed good effects for each activity. In addition, the isolated compounds were successfully quantified using a validated quantitative HPLC analysis method. As a result, the isolated compounds were suggested as standard marker compounds for Mongolian Shilajit. Also, we proved that the original material of Mongolian Shilajit is a lichen named Xanthoparmelia somloensis (Gyel.) Hale using HPLC-UV, ultra-high-performance liquid chromatography-electrospray ionization/hybrid linear trap-quadruple-orbitrap-high-resolution mass spectrometry (UHPLC-ESI/LTQ-HRMS).

A novel comparative chromatographic research of secoiridoid glycosides in two species of centaury herb

The aim: A novel comparative analysis of the secoiridoid glycosides composition in Centaurium erythraea Rafn. and Cеntaurium pulchellum (Sw.) Druce has being described. Swertiamarin has been chosen as an active marker and its comparative quantitative estimation in two species of the raw material has been carried out by HPLC method. Materials and methods: The quantitative HPLC analysis of swertiamarin was conducted using a chromatographic column ACE 5 C18. Methanol and 0,5 % acetic acid aqueous solution were used as mobile phases; the chromatographic procedure was carried out in a gradient mode. Results: The content of swertiamarin in the C. pulchellum herb ranges from 2.51 to 3.07 mg/100 mg. In that time swertiamarin content in C. erythraea herb varies and depends from 3.83 to 8.94 mg/100 mg. The content of swertiamarin in C. pulchellum herb is much lower than in C. erythraea herb. Taking into account this fact the possibility of using of C. pulchellum herb for medicine instead of C. erythraea herb must be proven additionally by biological experiments. Also it could be preliminary concluded that the most appropriate climate for swertiamarin accumulation in C. erythraea herb is located in Central and Eastern parts of Ukraine. Conclusions: It was discovered that the main representative of secoiridoid glycosides in C. erythraea is swertiamarin when in C. pulchellum Druce that are represented by both sweroside and swertiamarin. A more perspective raw material – C. erythraea herb, according to the content of swertiamarin, was chosen

Development of the technique for quality control of 1,3-bis(3,4-dicyanophenoxy)benzene by HPLC

Objectives. Determination of target products and byproducts is necessary for the quality control of phthalonitrile monomer synthesis as well as production scaling and performing related kinetic studies. High-performance liquid chromatography (HPLC) is a simple and affordable method for quantitative chemical analysis, which also verifies the quality of raw materials. The objective of this study was to develop an HPLC technique for determining the composition of the reaction mixture in the synthesis of 1,3-bis(3,4-dicyanophenoxy)benzene (DPB).Methods. Reversed-phase HPLC was used to quantitatively analyze the reaction mixture.Results. A simple and rapid method for the quantitative HPLC analysis of phthalonitrile monomers and their mixtures with reagents was developed. Reaction times and the accumulation of byproducts were also studied.Conclusions. The successful performance of the developed technique allows us to recommend it for practical applications. The results obtained for reactors of different sizes have good convergence, and DPB synthesis was successfully scaled up to intermediate scale equipment.

Comparison of Secondary Metabolite Contents and Metabolic Profiles of Six Lycoris Species

Quantitative HPLC analysis was performed on six different species of Lycoris herbs to investigate variation in phytochemical content, especially galantamine and phenylpropanoid-derived compounds. The contents of these compounds differed widely among the Lycoris species, with L. radiata and L. chinensis containing the lowest and highest galantamine contents, respectively. Specifically, the galantamine content of L. radiata was 62.5% higher than that of L. chinensis. Following L. radiata, L. sanguinea contained the next highest galantamine content, which was 59.1% higher than that of L. chinensis. Furthermore, a total of 12 phenylpropanoid-derived compounds were found in the different Lycoris species, where L. sanguinea, L. squamigera, and L. uydoensis had the largest accumulation of these compounds. The total phenylpropanoid content of L. sanguinea was the highest, while that of L. radiata was the lowest. Seven of the phenylpropanoid-derived compounds, rutin, quercetin, catechin, epicatechin gallate, chlorogenic acid, benzoic acid, and kaempferol, were dominant. L. sanguinea, L. uydoensis, and L. squamigera showed amounts of these seven compounds that were 5–6 times greater than those of the other species in the study. To the best of our knowledge, our results provide the most detailed phytochemical information on these species to date, which is valuable for future applications using these medicinal plants.

Establishing pinostrobin reference standard for quantitative analysis of the rhizomes of Boesenbergia pandurata

Boesenbergia pandurata Roxb. Schlecht. (Zingiberaceae), called ``Ngai bun'' in Viet Nam, is one of the Southeast Asian medicinal plants and its rhizomes are used primarily as a spice. This is a perennial, short-stemmed plant, formed by leaf sheaths and can grow up to 50 cm. The leaves are 7-11 cm wide and 25-50 cm long. Its rhizome surfaces are light brown in color, the inner rhizome is yellow, oval-shaped, and has a very aromatic odor. In folklore, Boesenbergia pandurata rhizomes are used as a spice for food processing. This plant contains pinostrobin as the major constituents. Previously showed that pinostrobin compound is the main ingredient together with a variety of biological activities such as antibacterial, inhibition of free radicals, ... Pinostrobin is necessary composition for the screening, testing, and quality evaluation of the rhizomes of B. pandurata species and others in the Zingiberaceae family. This research had conducted a reference standard of pinostrobin isolated from the rhizomes of B. pandurata had 99.26 % purity, which is reliable in medicinal testing. An HPLC method for pinostrobin determination was conducted and The quantitative HPLC analysis was validated for system suitability, selectivity, linearity ranges, and precision. Application of the process to investigate the preparation of extract shown that reflux extraction with ethanol obtained the highest pinostrobin content with 22.05 % in extracts and 2.89 % in dried rhizomes of B. pandurata.

CHEMOMETRICS ASSISTED VALIDATED HPLC METHOD DETECTING SEASONAL VARIANCES OF SECONDARY METABOLITES OF Camellia sinensis CONSIDERING THE ABIOTIC STRESSES

Climatic changes have great impact on the crops and agro-eco systems and such changes influences the concentration of secondary metabolites. IIT Kharagpur, India is a non-traditional tea growing zone where Tocklai Vegetative 25 variety was used as the research material. This study reports the development of a chemometrics assisted HPLC method validated as per ICH guidelines to explore the effect of seasonal variations in polyphenolics viz. catechins and methyl xanthenes like caffeine in fresh tea leaves and processed CTC black tea prepared from them. Further study was done on the variances amongst the concentration of secondary metabolites and abiotic stress factors. Good resolutions of secondary metabolites were obtained using 92% of 0.2% acetic acid and 8% of acetonitrile as the mobile phase, with a flow rate of 1 mL/ min, injection volume of 20 µl, PDA detector was set at 200-600 nm and chromatograms were recorded at 274 nm. Results of quantitative HPLC analysis have clearly shown that highest yield of catechins and caffeine were observed in fresh tea leaves plucked during spring (24.3ºC temperature and average rainfall of 34 mm) and also the processed black tea made from it, followed by tea leaves plucked during monsoon (28.8 ºC temperature and 282 mm rainfall) and processed black tea prepared from it. The lowest concentrations of secondary metabolites were found in leaves plucked during autumn (26.2 °C temperature and 132 mm rainfall) and the processed tea prepared from it. The developed quantitative HPLC method showed an inter day precision of 0.3, intraday precision of 0.2, repeatability value of 0.31, ruggedness value of 0.33 and robustness value of 0.2. Considering temperature and rainfall as abiotic stress factors, highest total polyphenolic content was obtained during spring and lowest in autumn. From our experimental findings, the fresh tea leaves of spring season and also the processed black tea prepared from it showed higher yield of catechins.

Chemical Analysis and Cytotoxic and Cytostatic Effects of Twelve Honey Samples Collected from Different Regions in Morocco and Palestine

The aim of this in vitro study is to characterize the phenolic compounds of twelve honey samples collected from different locations in Palestine (H1-6) and Morocco (H7-12) and to evaluate their cytotoxic and cytostatic effects in cells from the human colorectal carcinoma cell line HCT-116 and breast cancer cell line MCF-7. Quantitative HPLC analysis revealed nine phenolic compounds in three Moroccan honey samples, namely, syringic acid, tannic acid, caffeic acid, ferulic acid, coumaric acid, gallic acid, rosmarinic acid, epicatechin, and pyrogallol. Syringic acid, abundant in numerous types of honey with strong antioxidant capacities, was present at values ranging between 0.10 mg/100 g and 1.24 mg/100 g of Daghmos (H11) and Kabbar (H10) samples, respectively. No significant reductions in cell viability were observed in both cell lines treated with the Palestinian samples as measured with MTT assay. Significant cytostatic effects were after treatment of HCT cells with Morar honey H1 with IC50 of 1789 μg/ml. Three Moroccan samples, H7 (Zaâtar), H9 (Bochnikha), and H10 (Kabbar), showed slight, but significant cytostatic effects in HCT cells. A strong correlation was observed between cytostatic activity of MCF cells and antioxidant content (phenols, flavonoids, and flavonol). Furthermore, a strong negative correlation was detected between the cytostatic activity in HCT cells and the contents of syringic acid (r= -0.756) and tannic acid (r= -0.610). These results indicate that the traditionally known anticancer effects of honey might be mediated in part through cytostatic effects.