A green approach for metoclopramide quantification in pharmaceutical products: new HPLC and spectrophotometric methods.

Green spectrophotometric and HPLC methods have been developed for the quantification of metoclopramide. In the spectrophotometric method, it was determined by direct absorbance measurement at 273 nm wavelength using ultrapure water as solvent. The Extend C18 column was used for the HPLC method. The mobile phase system consisted of a combination of ethanol and formic acid solution (pH 2.0; 30:70 v/v). Isocratic elution was applied and the flow rate was set at 1.0 mL min-1. Metoclopramide was detected at 273 nm. The methods performed were economical, rapid, environmentally friendly, and simple, providing metoclopramide analysis within 5 min. The methods have been successfully applied in pharmaceutical products without matrix interference. The results of the application of the developed methods to pharmaceutical products were statistically compared and no significant difference was observed between the methods. In addition, the greenness assessment of the developed methods was performed using AGREE software. Our developed methods, based on the use of solvents such as ethanol and water, are proposed as a more environmentally and analyst-friendly option for the quantification of metoclopramide in pharmaceutical products than other methods currently in use.

Screening and classification of rosehip (Rosa canina L.) genotypes based on horticultural characteristics.

BACKGROUND: During the pandemic, the interest in colorful wild small fruits increased due to their positive effects on health. Also it has become very important to offer species with high nutritional value as fresh or processed products for human consumption due to increasing world population and decreasing arable land. In this context, we characterized the horticultural characteristics of 11 rosehip genotypes grown from seeds. RESULTS: Citric acid was determined as the main organic acid in all the genotypes investigated. The mean values of the organic acids obtained from all the genotypes were found to be as follows: citric acid (7177 mg L-1), malic acid (3669 mg L-1), tartaric acid (1834 mg L-1), oxalic acid (1258 mg L-1), carboxylic acid (631.9 mg L-1), shikimic acid (157.8 mg L-1), ascorbic acid (155 mg L-1), and acetic acid (20.9 mg L-1). Ellagic acid was the dominant phenolic compound (90.1 mg L-1 - 96.2 mg L-1) in all genotypes. The average values obtained from all genotypes for total phenolics, total flavonoids, and antioxidant activity were 37 261 mg GAE L-1, 526.2 mg quercetin L-1, and 93.6%, respectively. These characteristics had the lowest coefficients of variation, which indicated that all genotypes were similar regarding high biochemical with antioxidant effect. In addition, fruit width, fruit length, and fruit weight varied between 13.0 and 17.3 mm, 20.7 and 25.5 mm, and 1.4 and 2.7 g, respectively. CONCLUSIONS: The genotypes were categorized according to different purposes, such as suitability for wine production, making vinegar, etc. While the pomological characteristics were strongly positively correlated among themselves, they were generally found to be negatively correlated with the phytochemical characteristics. Categorizing genotypes according to different usage purposes can improve the agricultural and industrial application of rosehip and enhance their breeding efficacy.

Biological Activities of Galanthus fosteri Extracts: First Demonstration of the Interaction between Chlorogenic Acid and DNA Ligase by Molecular Docking.

Within the Amaryllidaceae family, the bulbous plant species Galanthus fosteri (G. fosteri) belongs to the Galanthus genus. Alkaloids with a broad variety of biological functions are typically found in the flora of this family. The G. fosteri plant's organs' antioxidant activity, antibacterial impact, and antimicrobial qualities were examined in this study. Total flavonoid contents (TFC) and total phenolic contents (TPC) of plant extracts were measured with spectrophotometric methods, and antioxidant activity was determined using the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging technique. The HPLC method was used to determine the phenolic compounds on a component basis. The antibacterial properties of the extracts were assessed using the Kirby-Bauer disc diffusion method, and the minimum inhibitory concentration method against the pathogens Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. Additionally, combination tests were performed between the extract and antibiotics. Leaf and stem extracts demonstrated greater antioxidant qualities than bulb extracts, despite the fact that extracts of plant organs did not exhibit appreciable levels of TPC, TFC, or antioxidant qualities. According to the HPLC analysis results, it was determined that chlorogenic acid was present in all of the extracts. In fact, it was determined that only chlorogenic acid was 8.02 (mg/10 g) in G. fosteri bulb peel, which has antimicrobial and antioxidant properties. A molecular docking study has demonstrated for the first time that the antibacterial effect of chlorogenic acid might be due to DNA replication inhibition.

[The Effect of Oleuropein on The Mitochondrial Membrane Potential and Generation of Reactive Oxygen Species on Leishmania tropica Promastigotes]. / Oleuropein'in Leishmania tropica Promastigotlari Üzerinde Mitokondri Membran Potansiyeli ve Reaktif Oksijen Türlerinin Olusumuna Etkisi.

Leishmania parasites, which are reported to be endemic in 98 countries around the world, infect humans as well as wild and domestic carnivores and small mammals, and are transmitted by sand flies (Phlebotomus, dwarf sandflies). It is reported that 350 million people are at risk and two million new cases are seen in the world every year. It has been reported that different drugs (topical paromomycin, oral miltefosine, ketoconazole, rifampin, and zinc) have been tried in studies especially in endemic regions in the treatment of cutaneous leishmaniasis, and response to treatment has been obtained at different rates. Today, the search for alternative treatments continues and many studies have been carried out for this purpose. For centuries, olive leaf extracts have been used to maintain health. Oleuropein has numerous health benefits, including antioxidant, antimicrobial, anti-inflammatory, antiatherogenic, anticarcinogenic, antiviral activities, cardio- and neuroprotective, hepatoprotective effects. The aim of this study was to determine and understand the mode of action of oleuropein, the cell death mechanisms caused by oleuropein in L.tropica promastigotes. In this study, the phenolic and flavonoid content of oleuropein was determined by HPLC method. The antioxidant capacity and the amount of oleuropein were determined. Afterwards, morphological and physiological (mitochondrial membrane potential, formation of reactive oxygen species, Annexin V binding) changes triggered by oleuropein in L.tropica promastigotes were investigated using flow cytometry. Our studies revealed that apoptotic properties such as mitochondrial dysfunction, production of reactive oxygen species, flip-flop action of phosphatidylserine could induce cell death in L.tropica promastigotes. It has been observed that oleuropein induced typical apoptotic morphological features in L.tropica promastigotes. Total phenolic content and total flavonoid content values of oleuropein extract were determined as 33 mg/g and 229 mg/g. The radical removal method was used to investigate the antioxidant capacity of methanol extracts against free radicals. Total antioxidant content of oleuropein extract was determined as 87%. In addition, the amount of oleuropein in the oleuropein extract was determined as 21. 1% by HPLC. The oleuropein dose that killed 50% of L.tropica promastigotes, that is the IC50 value, was detected as 46.6 µg/mL after 24 hours. It was observed that the parasites in the control group preserved their typical morphological features with a single nucleus, flagella, kinetoplast and narrow cell body at both 24 and 48 hours. It was observed that as oleuropein concentrations increased, the and kinetoplasts of L.tropica promastigotes could not be distinguished from each other, they moved away from the narrow cell body structure, they lost their flagella and turned into a round form, and they moved away from the typical form of the parasite. The percentage of Annexin V+ apoptotic cells was found to be 2.9 ± 0.4% in the untreated control group, and 38.1 ± 6.9% in the oleuropein-treated group. Polarization in the mitochondrial membrane of healthy promastigotes caused an approximately 1.7-fold change in the direction of depolarization in oleuropein-treated promastigotes. According to these findings, oleuropein triggered mitochondria-related death in L.tropica promastigotes. Moreover, 1.4 ± 0.2 fold increase in reactive oxygen species production was detected in oleuropein-treated promastigotes compared to the untreated control group. Comparisons between groups were made using the independent sample t test method. In conclusion, phenolic compounds of olive leaf extract oleuropein induced apoptotic cell death in L.tropica promastigotes. Our results support that olive products such as oleuropein may have anti-parasitic effects.

Screening of Naturally Grown European Cranberrybush (Viburnum opulus L.) Genotypes Based on Physico-Chemical Characteristics.

It has become very important to offer species with high nutritional value as fresh or processed products for human consumption in their daily diet for balanced nutrition. In the scope of this study, 15 naturally grown European Cranberry bush (ECB) genotypes that naturally grown were characterized in terms of horticultural characteristics. Fruit length, fruit width, fruit weight, the number of fruits per each cluster and cluster weight were determined within the ranges of 8.78−10.96 mm, 7.93−10.84 mm, 0.21−0.70 g, 31−121, and 7.70−66.67 g, respectively. Ranking of the average values of examined organic acids obtained from all genotypes found as; malic acid (11,419 mg L−1) > citric acid (1926 mg L−1) > ascorbic acid (581 mg L−1) > oxalic acid (561 mg L−1). Total phenolic content (TPC) and total flavonoid content (TFC) were found at high levels in ECB with 2922−3475 mg gallic acid equivalent (GAE) L−1 and 1463−3163 mg quercetin equivalents (QE) L−1, respectively. While pomological characteristics were found to be highly positive correlated with each other, they were negatively correlated with chemical properties. Low pH was found to be an important parameter to obtain higher amounts of phytochemicals such as TPC, TFC, organic and phenolic acids correlated with strong antioxidant effects. The obtained results will be useful for both germplasm enrichment and cultivation.

Comparison of HPLC and UV Spectrophotometric Methods for Quantification of Favipiravir in Pharmaceutical Formulations.

There is no currently successful method to treat Covid-19 infection. Nevertheless, previously licensed pharmaceuticals to treat other virus infections are used on an off-label basis either alone or in combination. One of them is favipiravir. Favipiravir, also known as favilavir, is an antiviral drug that is active against many viruses. Spectrophotometric and liquid chromatographic methods have been developed and validated for the quantitative determination of favipiravir in pharmaceutical formulations. Chromatographic method has been performed using reverse-phase technique on a C-18 column with a mobile phase consisting of sodium acetate solution (pH adjusted to 3.0 with glacial acetic acid) and acetonitrile (85:15, v/v) at 30 oC. The mobile phase flow rate was 1.0 mL min-1. For the determination of favipiravir, UV spectrum has been recorded between 200 and 800 nm using deionized water as solvent and the wavelength of 227 nm has been selected. Both methods have been validated in terms of their specificity, linearity, limits of detection and quantification, precision, accuracy, and robustness. Both methods have demonstrated good linearity, precision and recovery. No spectral and chromatographic interferences from the tablet excipients were found in spectrophotometric and liquid chromatographic methods. In both methods, correlation coefficients were greater than 0.999 within a concentration range of 10-60 mg mL-1 using spectrophotometry and chromatography. Intra-day and inter-day precision were observed with low relative standard deviation values. The accuracy of the methods were within the range 99.57-100.10% for LC and from 99.83-100.45% for UV. Therefore, both methods gave the most reliable outcomes for the determination of favipiravir in pharmaceutical formulation.

Extraction, development and validation of HPLC-UV method for rapid and sensitive determination of colchicine from Colchicum autumnale L. Bulbs.

Colchicum autumnale L. also known as the autumn crocus, contains colchicine alkaloid having antifungal properties. The tuber of this plant is rich in terms of colchicine. In this research an ultrasound-assisted extraction (UAE) method was optimized for the extraction of colchicine from Colchicum autumnale L. bulbs before high-performance liquid chromatography with UV detection (HPLC-UV). Optimization of various extraction parameters was performed using response surface methodology (RSM) to evaluate the maximum colchicine yield from Colchicum autumnale L. bulbs. The Box-Behnken design (BBD) and RSM were used to investigate the effect of three key parameters (extraction time (20-60 min), extraction temperature (40-80 °C) and ultrasound power (500-700 W) on extraction efficiency. The variance analysis suggested that the dependent response variable of yield of colchicine may be expressed by a quadratic polynomial model. The optimal theoretical extraction conditions were found to be an ultrasonication power of 602.4 W, an extraction time of 42 min and a temperature of 64 °C. Under these conditions, the optimum foreseen yield was 0.237%. The experimental colchicine yield obtained by following the optimized conditions was found to be 0.238%. These values are very well compatible with each other.

Development and validation of a capillary zone electrophoretic method for rapid and sensitive determination of galanthamine: Application in plant and pharmaceuticals.

Galanthamine is a second-generation cholinesterase inhibitor that has begun to be used in the treatment of Alzheimer's disease. In the presented research, a simple, accurate, precise and cost-effective capillary zone electrophoretic (CZE) method was used for the qualitative and quantitative estimation of Galanthamine from bulbs, leaves and fringes of Leucojum aestivum L. (summer snowflake) grown in Turkish habitats (Black Sea Region) and pharmaceutical dosage forms by capillary zone electrophoresis with direct UV detection form. Ultrasonic assisted extraction (UAE) and response surface methodology (RSM) were used to estimate optimum experimental conditions on the content extraction of Leucojum aestivum L. Metoprolol was used as a suitable internal standard. A linear relationship between the ratio and concentrations of Galanthamine in the range of 0.25µgmL-1-15.00µgmL-1was determined with a regression coefficient of 0.9996, for which the limit of detection (LOD) and limit of quantitation (LOQ) were 0.027 and 0.081µgmL-1, respectively. The high percentage recovery results showed that the matrix effect did not influence the developed method for analysis of pharmaceutical preparations. Validation parameters were carried out according to the guidelines of the International Conference for Harmonization (ICH). This method also allows for a number of cost- and time-saving benefits and can be readily employed for the analysis of plants and pharmaceutical formulations. The method can be used in industries for the determination of Galanthamine to analyze the quality of extraction and formulation without interference.

Optimization of Ultrasound-Assisted Extraction of Morphine from Capsules of Papaver somniferum by Response Surface Methodology.

In this study, amount of morphine from poppy capsules (Papaver somniferum) was investigated using ultrasonic assisted extraction (UAE). Response surface methodology was used to estimate effective experimental conditions on the content extraction of poppy capsules. For this purpose, solvent/solid ratio (10-20 mL/500 mg sample), pH (1-13), time (30-60 min), and temperature (30-50°C) were chosen as experimental variables. The affected response is extraction recovery values for morphine from poppy straw. For interpreting the relationship between experimental factors and response, a design table was established with combinations of three different concentrations levels of this compound in 29 trials. The second order quadratic model gave a satisfactory description of the experimental data. In our study, R-Squared (0.96), Adj-R-Squared (0.92), and Pred R-Squared (0.78) values for extraction yield display good accuracy of the derived model. The predicted optimal conditions for the highest morphine level (3.38 mg morphine/500 mg-sample) were found at 19.99 mL solvent/500 mg solid ratio, 59.94 min extraction time, 1.10 pH, and 42.36°C temperature. In the optimal extraction conditions, the experimental values are very close to the predicted values. Consequently, the response surface modeling can be achieved sufficiently to predict extraction yield from poppy straw by ultrasound assisted extraction.

Radiolabeling of morphine with (131)i and its biodistribution in rats.

This study was conducted to determine the possible radiopharmaceutical potential of morphin labeled with (131)I. Morphine was extracted from dry capsules of the opium poppy (Papaver somniferum L.), purified by high-performance liquid chromatography, and characterized with nuclear magnetic resonance and infrared spectroscopy. The purified compound was labeled with (131)I. Male Albino Wistar rats (18) were used for receptor blockage and unblockage biodistribution studies. Tissue distribution studies showed that radiolabeled morphine had higher uptake in lung, liver, small intestines, large intestines, and stomach than the other tissues. The highest uptake of radiolabeled compounds in rats' brain was found to be in the midbrain and hypothalamus. After receptor blockage with morphine, uptake of (131)I-morphine decreased in the lungs, liver, kidney, testis, prostate, spinal cord, cerebellum, hippocampus, striatum, and temporal cortex with respect to receptor unblockage studies of rats. This study concludes that the labeling yield of (131)I-morphine was high, high amount of (131)I-morphine was found in the hypothalamus, and (131)I-morphine has enough stability for diagnostic scanning.

Human recombinant anti-La (SS-B) autoantibodies demonstrate the accumulation of phosphoserine-366-containing la isoforms in nucleoplasmic speckles.

Using the recombinant La (SS-B) protein or a phosphorylated peptide derived thereof 27 La-specific human recombinant autoantibodies were selected from anti-La-positive systemic lupus erythematosus and systemic sclerosis patient-derived combinatorial phage display antibody libraries. Binding of these anti-La antibodies to various isoforms of the La protein present in normal and apoptotic cell extracts was analysed by Western blotting. Twenty-four of the selected antibodies recognize most, if not all isoforms of La, whereas three are exclusively reactive with the protein phosphorylated at serine-366. Sequence analysis of the selected antibodies showed a restricted spectrum of diversity in their VH germline gene usage. Remarkably, the recombinant antibodies recognizing exclusively the phosphoserine-366-containing isoform of La displayed a spleckled nucleoplasmic staining pattern in immunofluorescence analysis of HeLa and HEp-2 cells. This pattern differed markedly from those obtained with anti-La antibodies recognizing all isoforms of the La protein. Colocalization experiments with marker antibodies for spliceosomal UsnRNPs and RNA polymerase III subunits revealed that the anti-phosphorylated La antibodies stain the same nucleoplasmic speckles as anti-UsnRNP antibodies. In contrast to anti-UsnRNP antibodies the anti-phosphorylated La antibodies did not stain the Cajal bodies. In addition, no colocalization of phosphorylated La with RNA polymerase III was observed. Potential functional implications of the accumulation of phosphorylated La in nucleoplasmic speckles are discussed.