High-performance Liquid Chromatography Technique Research Articles

DETERMINATION OF THE OPTIMAL CONDITIONS FOR LEVAN PRODUCTION FROM THE LOCAL ISOLATE OF Bacillus subtilis Strain AE77

Background: Levan is a promising safe, renewable, and environmentally friendly exocellular homopolysaccharides. It has recently gained increasing research interest because of its significant impact on biotechnology applications due to its biocompatibility and ability to biodegradability. Aim: The study aimed to determine some of the optimal cultural conditions for levan production by studying many parameters affecting production by local isolation Bacillus subtilis (B. subtilis Strain AE77). Methods: B. subtilis Strain AE77 It was obtained from the Department of Food Science/College of Agricultural Engineering Sciences as a local levan-producing isolate isolated from rotten sweets. The Mineral Salts Broth (MSB) medium was approved to study the optimized conditions for levan production. Then precipitation of levan from the fermentation medium. Then the identification of levan with High-Performance Liquid Chromatography (HPLC) technology by estimating the retention time of levan, compared with the standard model produced from the bacterial isolate Erwinia herbicola (E.herbicola). Besides, each of the standard solutions of sugars (fructose, glucose, and sucrose). Results: The study results proved that the polysaccharide produced from this isolate is pure and consists of fructose and is a homopolysaccharide with the HPLC technique. The best levan production was from B. subtilis Strain AE77 using a modified MSB medium of 20% sucrose in a 48 hours incubation period. An inoculum volume of 1×108 cell/ml at a concentration of 2% at an incubation temperature of 35 °C and pH = 7.0 and agitation speed of 150 rpm reached 4.80 g/100ml. Conclusion: HPLC was used to separate, identify and quantify both B.subtilis Strain AE77 and standard levan produced from the bacterial isolate E. herbicola, as well as standard fructose, glucose, and sucrose, respectively. The results showed that levan is a homopolysaccharide consisting of one type of monomer (the fructose sugar units). Conditions for levan production were determined by isolating B. subtilis Strain AE77 in a sucrose medium. The results showed that the optimal medium for production contained 20% sucrose without any nitrogen source, pH7.0, and inoculum volume 1x108 cell/ml at a concentration of 2% at a temperature of 35 °C agitation speed 150 rpm, and incubation period of 48 hours.

Characterization of 6-Gingerol for In Vivo and In Vitro Ginger (Zingiber officinale) Using High Performance Liquid Chromatography

Ginger (Zingiber officinale Rosco) belonging to the family Zingiberaceae is one of the world’s most important spices and produces a pungent, aromatic rhizome that is valuable all over the world. Qualitative and quantitative analysis of 6-gingerol in different parts (in vivo and in vitro) of Zingiber officinale using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) have been performed. Data of TLC showed spots having identical Rf value (0.15), according to the synthetic standards of 6-gingerol in all samples extract. 6-gingerol was detected in all extracts of different parts of ginger derived from in vivo and in vitro culture conditions. Quantitative determination of 6-gingerol using HPLC technique was carried out. Comparing with the peaks of 6-gingerol in synthetic standards, in vivo rhizomes and in vitro cultures of different ginger parts was showed similar UV spectra characteristics. The quantity of 6-gingerol in rhizomes (in vivo and in vitro) and in vitro microrhizomes (45.37; 42.64; 28.11 mg/g respectively), were showed a higher value than that of in vitro calli, shoots and roots (7.89; 7.46; 6.40 mg/g respectively).

Gastroprotective effect of hydroalcoholic extract from Agaricus blazei Murill against ethanol-induced gastric ulcer in mice

Ethnopharmacological relevanceThe use of mushrooms in medicine is quite old and the first report about the use of genus Agaricus in treatment of ulcers occurred in Byzantine period. This mushroom is widely consumed as food, tea, food supplements, as well as nutraceutical and cosmeceutical applications, being cultivated and appreciated in several countries such as Brazil, Korea, Japan and China. Aim of the studyThis study aimed to characterize the chemical profile and the potential gastroprotective effect of hydroalcoholic extract from Agaricus blazei Murill (HEAb). Materials and methodsThe extract was chemically characterized by elemental analysis, UPLC-QTOF-MSE, Nuclear Magnetic Resonance (NMR) and high-performance liquid chromatography (HPLC) techniques to elucidate the metabolites present in the extract. The quantification of phenolic compounds and the in vitro antioxidant activities were performed and the gastroprotective effect of this extract was evaluated against ethanol-induced gastric ulcer model. HEAb was administered by gavage at 5, 25 and 50 mg kg−1 and N-acetylcysteine at 300 mg kg−1 (positive control). Furthermore, the pathways of nitric oxide (NO), Cyclic Guanylate Monophosphate (cGMP), prostaglandins (PGs) and the involvement of ATP-sensitive K+ Channels were modulated. ResultsMannitol, malic acid, pyroglutamic acid, L-agaritine and L-valine were putatively identified by UPLC-QTOF-MSE in HEAb. In addition, it was possible to identify mannitol by the intense signals in the NMR spectra, being still quantified as the main compound in the extract by HPLC. The contents of total phenols and flavonoids corroborated with the good antioxidant activity of HEAb. This study observed that HEAb at 25 and 50 mg kg−1 had gastroprotection effect demonstrated by the reduction of histopathological parameters and the reduction of mastocytosis in the stomach of mice. ConclusionsIn this study was possible to conclude that HEAb has gastroprotective effect related to the involvement of NO and PG pathways in the ethanol-induced gastric ulcer model in mice.

Development of Urinary Assay Methods for the Estimation of Paracetamol Glucuronide and Paracetamol Sulphate in Preterm Neonates with Patent Ductus Arteriosus

Aims: This study aimed to develop a high-performance liquid chromatography (HPLC) technique for estimating paracetamol glucuronide and paracetamol sulphate in the urine samples of preterm neonates. Background: Validated methods exist for estimating the principal metabolites of paracetamol in older children and those with liver disease. Here, we have developed and validated a simple technique for estimating the same in urine samples of preterm neonates. Objective: The study aims to develop and validate a simple, reliable, and accurate HPLC technique for estimating urinary paracetamol glucuronide and paracetamol sulphate metabolites. Methods: Preterm neonates of either sex diagnosed with patent ductus arteriosus (PDA) receiving paracetamol intravenously at the dose of 15 mg/kg every six hours were recruited. We ran the samples under standardized chromatographic conditions and using various dilutions of the calibration standards. Measures of assay selectivity, linearity, accuracy, and precision were estimated. Results: We observed that the peaks for paracetamol glucuronide and paracetamol sulphate were distinguished from those of the drug-free urine samples. The results for both metabolites revealed good reproducibility, with a percent coefficient of variation (% CV) of 4.3 and 4.9 for the slope for paracetamol glucuronide and paracetamol sulphate, respectively. Similarly, we observed good linearity, as indicated by the correlation coefficients of 0.99 for the metabolites. The validation assays revealed that the method is linear, accurate, and precise over the defined concentration ranges. Conclusion: We demonstrated that HPLC has good accuracy, reliability, and precision, and it can be used for estimating the principal metabolites from urine samples in neonates for defining the ontogeny of conjugation enzymes and in paracetamol overdose.

The potential of common duckweed (Lemna minor) in phytoremediation of phenanthrene and pyrene

The distribution of polycyclic aromatic hydrocarbons (PAHs) as a group of toxic and persistent aromatic pollutants in the environment is rapidly enhancing. These compounds have adverse impacts on the health of living organisms. Hence, in the present study, we investigated the potential of duckweed (Lemna minor) as an aquatic plant species for uptake, accumulation, and biodegradation of phenanthrene and pyrene under controlled conditions. L. minor plants were treated with 10 and 20 mg L-1 concentrations of phenanthrene and pyrene at the experimental duration of ten days. According to the results obtained, the toxicity of phenanthrene and pyrene contaminants on L. minor was influenced by the different initial PAHs concentrations. An increase in phenanthrene and pyrene concentration significantly decreased all studied growth parameters such as fresh weight, dry weight, and RFN and also photosynthetic pigment contents of the plant. Phenanthrene and pyrene concentrations were measured using high-performance liquid chromatography (HPLC) technique after 10 days of exposure to the PAHs. The results revealed that L. minor species could bioaccumulate effectively both typical PAHs. Furthermore, the gas chromatography-mass spectroscopy (GC/MS) technique explained the biological degradation of phenanthrene and pyrene by L. minor in the present research, and accordingly, several intermediate by-products were identified.

Extraction of Bioactive Compounds from Ulva lactuca

Macroalgae Ulva lactuca, has been employed as a natural source for the production of extracts with potent bioactivity. The biochemical characterization showed that the macroalgae biomass contains a remarkable amount of the polysaccharide Ulvan (49.9 wt%) which is a valuable chemical compound well known for its benefits in human health. Four nontoxic solvents, water, ethyl acetate, ethanol, and an ethanol/water mixture (70:30 v/v) were examined for their recovery efficiency of total carotenoid and phenolic contents. Experimental results showed that the aqueous mixture of ethanol was the most efficient solvent in the recovery of bioactive compounds with extraction yield of 10–15% dw. The effect of extraction parameters, namely time, temperature, and the ratio of biomass to solvent, on the carotenoid and phenolic compounds’ content, antioxidant activity, and extraction yield, was investigated, using the ethanol/water mixture as a solvent. The extract obtained under 60 °C, 3 h of extraction time and 1:10 biomass to solvent mass ratio showed the highest antioxidant activity. This extract maintained its antioxidant capacity almost stable for five days of storage under cool and dark conditions. Finally, specific phenolic and carotenoid compounds in the U. lactuca extracts were identified using the High-Performance Liquid Chromatography (HPLC) technique.

Greenness estimation of chromatographic assay for the determination of anthracycline-based antitumor drug in bacterial ghost matrix of Salmonella typhimurium

The determination of an anthracycline-based antitumor drug, doxorubicin (DOX), in the bacterial ghost (BG) matrix of Salmonella typhimurium and commercial injectable dosage form was performed using a rapid, sensitive, and environmentally greener “high-performance liquid chromatography (HPLC)” assay. DOX was determined chromatographically on a Nucleodur (150 mm × 4.6 mm) RP C18 column with a particle size of 5 μm. The greener solvent system consisted of ethanol, methanol, and acetic acid (50:49:1 percent v/v/v), which was delivered with a flow rate of 1.0 mL/min. DOX was detected at a wavelength of 480 nm. With a determination coefficient value of 0.9992, the suggested HPLC assay was linear in the range of 1–100 μg/mL. In addition, the suggested HPLC assay for DOX analysis was rapid (Rt = 2.19 min), accurate (percent recoveries = 98.12–101.55), precise (percent CV = 0.87–1.97), and sensitive. The applicability of suggested HPLC technique was demonstrated by measuring DOX in S. typhimurium BG matrix and commercial injectable dosage form. DOX recovery in commercial injectable dosage form was calculated to be 101.02 percent. The amount of DOX in BG matrix was determined to be 48.51 μg/mL. Using the AGREE metric technique, the overall analytical GREEnness (AGREE) score for the proposed HPLC assay was calculated to be 0.79, indicating an outstanding greenness profile. These findings suggested that the proposed HPLC assay might be used to analyze DOX in BG matrices and commercial dosage forms on a regular basis.

A Kinetic Study of Photocatalytic Degradation of Phenol over Titania–Silica Mixed Oxide Materials under UV Illumination

A set of titania–silica mixed oxide materials were prepared by a cosolvent-induced gelation method using ethanol and toluene as solvent and cosolvent, respectively. These materials were extensively characterized by utilizing several characterization techniques and assessed for phenol degradation under UV illumination. The degradation of phenol follows first-order kinetics, and fragmented products formed during the phenol degradation were qualitatively identified by using high performance liquid Chromatographic (HPLC) and atomic pressure chemical ionization mass spectroscopic (APCI-MS) techniques. The complete mineralization of phenol was further evidenced by the measurement of the total organic contents that remained in the solution after irradiation. The pore diameter of the materials was found to be the key factor for phenol degradation, whereas surface area and pore volume play a role among the mixed oxide materials. In addition, in the mixed oxide system there was an inverse correlation obtained with the particle size of the materials and the degradation efficiency. The smaller particle size of titania in the mixed oxide material was found to be a requirement for an effective degradation of phenol.

Relationship between Oxidative Stress and the Blood Iron Concentration and Antioxidant Status in Major ß-thalassemia in Iraq.

Reduction or total lack of beta-globin chains caused by a congenital disease called ß-thalassemia major is one of the lives threatening diseases. Patients who suffer from ß-thalassemia need a repeated blood transfusion for survival. The repeated blood transfusion in ß-thalassemia patients may cause oxidative stress and tissue injury due to iron overload, altered antioxidant enzymes, and other essential trace element levels. The current study aimed to investigate the correlation of oxidative stress with serum trace element levels and antioxidant enzyme status in ß-thalassemia major patients. A total of 130 serum samples were obtained from ß-thalassemia major patients (n=100; 50 males and 50 females) and healthy individuals (n=30; 15 males and 15 females). Hematological parameters were measured on both groups by a comprehensive blood test that included the amount of hemoglobin Hb, packed cells volume, number of red blood cells, mean corpuscular volume ratio, mean corpuscular hemoglobin ratio, mean corpuscular hemoglobin concentration, red cell distribution width, white blood cells, and platelets counts. All of these blood parameters showed a clear decrease in thalassemia patients, except for red blood cells and platelets counts, which demonstrated a significant increase. The highest significant mean for iron in males and females were 233.768 and 219.150 µgm\dL in patients, respectively, while the mean level of iron significantly reduced in the control group (113.40 and 103.33 µgm\dL in males and females, respectively). The results indicated a significant decrease in uric acid in males and females in the patient group (41.042 and 40.582 mg\L in males and females, respectively), compared to the control group (53.866 and 43.60 mg\L in males and females, respectively). Allantoin concentration was detected by high-performance liquid chromatography technique, the results of which showed that the highest values in patients were 62.822 and 25.480 mg\L in males and females, respectively, compared to the control group 2.342 and 1.481 mg\L in males and females, respectively. Superoxide dismutase concentration decreased in patients (129.635 and 111.848 U\mL in males and females, respectively), compared to the control group (208.623 and 190.413 U\ml in males and females, respectively).

Protein extract of kenaf seed exhibits anticoagulant, antiplatelet and antioxidant activities

Objective: To explore the anticoagulant, antiplatelet and antioxidant activities of protein extract of kenaf seed (PEKS). Methods: Sodium dodecyl sulphate polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography techniques were employed for protein characterization. Antioxidant activity of PEKS was assessed using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The protective effect of PEKS on sodium nitrite (NaNO2) induced oxidative stress was evaluated using the in vitro red blood cell model, while the effect of PEKS on diclofenac-induced oxidative stress was examined in vivo in rats. Platelet-rich plasma and platelet-poor plasma were used for anticoagulant and antiplatelet activities of PEKS. Results: PEKS revealed similar protein bands on SDS-PAGE under reduced and non-reduced conditions. Several acidic proteins were present in native PAGE. PEKS showed antioxidant properties by scavenging DPPH with an IC50 of 24.58 μg. PEKS exhibited a protective effect on NaNO2 induced oxidative stress in red blood cells by restoring the activity of stress markers. In addition, PEKS alleviated diclofenac-induced tissue damage of the liver, kidney, and small intestine. PEKS showed an anticoagulant effect in both in vivo and in vitro experiments by enhancing normal clotting time. PEKS did not affect prothrombin time but increase activated partial thromboplastin time. Furthermore, PEKS inhibited adenosine diphosphate and epinephrine-induced platelet aggregation. Conclusions: PEKS protects tissues from oxidative stress and exhibits antithrombotic activity.

Analytical Method Development And Validation Parameters Of Drug Ivermectin

An accurate, easy, detailed, selective and fast RP-HPLC stability representative technique was developed and validated for assessment of ivermectin in tablet dosage form. The Reverse Phase High Performance Liquid Chromatographic technique was developed for routine quantification of ivermectin in laboratory prepared mixtures as well as in combined dosage forms. The chromatographic separation was accomplished with INERTSIL C-18 ODS 250×4.6mm, 5µm particle size column along with acetonitrile and methanol as the mobile phase at a flow rate of 1ml/min. Quantification was completed by using a UV detector at 245 nm and the run time was 10 minutes. The retention time was found to be 4.198 min for ivermectin. The linearity was observed in the range of 1-32µg/ml with correlation coefficient r= 0.9798. The % RSD for intraday and interday precision was 1.352 and 1.589 respectively. The LOD and LOQ values were found to be 2.93 and 8.79, respectively. The system suitability parameters for ivermectin such as theoretical plates and tailing factor were found to be 129.949 and 2.0, respectively. Robustness was also studied and there was no significant variation in the system suitability of the analytical method by incorporating small changes in experimental parameters. The technique has been validated for linearity, precision, accuracy and other parameters as approved by ICH guidelines. The results obtained by RP- HPLC methods are found to be fast, detailed, selective and accurate. Therefore, proposed analytical method can be used for regular analysis of ivermectin in injection, tablet and other formulations.

A sustainable approach on biomining of low-grade bauxite by P. simplicissimum using molasses medium.

In order to find a sustainable and low-cost alternative route to the traditional recovery of aluminum, the filamentous fungus Penicillium simplicissimum was evaluated for aluminum recovery from low-grade bauxite ore. The oat-agar medium was carefully chosen as the foremost solid medium for fungal sporulation due to lower cost, ease in preparation, and high spore production in a short incubation time. To examine the acid production capability in submerged fermentation, P. simplicissimum was inoculated in a medium augmented with glucose and molasses as an energy source. High-performance liquid chromatography (HPLC) technique was used for the determination of the produced organic acids. Three different bioleaching approaches were evaluated using 1% bauxite pulp density. The culture containing P. simplicissimum spores grown in a medium supplemented with molasses leached 86.6% Al in the direct two steps on the fifth day, 56.5% in the direct one step on the fourth day, and 71.7% in the indirect bioleaching on the fourth day, while in the controlled sterile flasks, Al leaching was almost negligible. A maximal amount of Al was leached by the fungal strains using low-cost molasses as a substrate.

Correlation of whole blood hydroxychloroquine concentration with cutaneous lupus erythematosus and factors associated with it: First multicenter, cross-sectional analysis in Malaysia.

Hydroxychloroquine (HCQ) is the first-line systemic treatment for cutaneous lupus erythematosus (CLE). Whole blood HCQ concentration (WBHCQ) was found to correlate with CLE severity among Caucasians. However, studies on Asians are scarce. We aim to explore the relationship of WBHCQ with CLE disease activity among multi-racial Malaysians and the factors associated with WBHCQ. A cross-sectional study targeting patients with CLE was conducted from 1 June till 30 November 2019. Disease activity was assessed using Cutaneous Lupus Erythematosus Disease Area and Severity Index - Activity Score (CLASI-AS). Blood was analyzed for WBHCQ concentration using a high-performance liquid chromatography technique. Statistical analysis was done using R studio version 1.2.1335. A total of 88 subjects (male:female, 4.5:1) with a median age of 41years old were recruited. The median duration CLE was 5years. The majority had acute cutaneous lupus (n=45, 51.1%). The median WBHCQ was 946.8ng/mL. Indians were found to have the highest WBHCQ (median±interquartile range [IQR], 1515.4±1494.8ng/mL). Males had a lower WBHCQ (median±IQR, 733.5±573.8ng/mL) than females (995.5±925.1ng/mL). However, no statistically significant association between race and sex with WBHCQ was demonstrable (p=0.247, p=0.066). No correlation was demonstrated between WBHCQ and CLASI-AS (r=-0.02, p=0.851). A positive correlation was found between HCQ dosage (ideal bodyweight) and WBHCQ (r=0.24, p=0.027). No other factors were found associated with WBHCQ. Indians and females were observed to have higher WBCHQ; however, no significant correlation was identified. Further study is required to confirm the finding.

Identification of Antibacterial Components in the Methanol-Phase Extract from Edible Herbaceous Plant Rumex madaio Makino and Their Antibacterial Action Modes.

Outbreaks and prevalence of infectious diseases worldwide are some of the major contributors to morbidity and morbidity in humans. Pharmacophageous plants are the best source for searching antibacterial compounds with low toxicity to humans. In this study, we identified, for the first time, antibacterial components and action modes of methanol-phase extract from such one edible herbaceous plant Rumex madaio Makino. The bacteriostatic rate of the extract was 75% against 23 species of common pathogenic bacteria. The extract was further purified using the preparative high-performance liquid chromatography (Prep-HPLC) technique, and five separated componential complexes (CC) were obtained. Among these, the CC 1 significantly increased cell surface hydrophobicity and membrane permeability and decreased membrane fluidity, which damaged cell structure integrity of Gram-positive and -negative pathogens tested. A total of 58 different compounds in the extract were identified using ultra-HPLC and mass spectrometry (UHPLC-MS) techniques. Comparative transcriptomic analyses revealed a number of differentially expressed genes and various changed metabolic pathways mediated by the CC1 action, such as down-regulated carbohydrate transport and/or utilization and energy metabolism in four pathogenic strains tested. Overall, the results in this study demonstrated that the CC1 from R. madaio Makino are promising candidates for antibacterial medicine and human health care products.

Carbon-11 Radiotracing Reveals Physiological and Metabolic Responses of Maize Grown under Different Regimes of Boron Treatment.

In agriculture, boron is known to play a critical role in healthy plant growth. To dissect the role of boron in maize metabolism, radioactive carbon-11 (t½ 20.4 min) was used to examine the physiological and metabolic responses of 3-week-old B73 maize plants to different levels of boron spanning 0 mM, 0.05 mM, and 0.5 mM boric acid (BA) treatments. Growth behavior, of both shoots and roots, was recorded and correlated to plant physiological responses. 11CO2 fixation, leaf export of [11C]-photosynthates, and their rate of transport increased systematically with increasing BA concentrations, while the fraction of [11C]-photosynthates delivered to the roots under 0 mM and 0.5 mM BA treatments was lower than under 0.05 mM BA treatment, likely due to changes in root growth. Additionally, solid-phase extraction coupled with gamma counting, radio-fluorescence thin layer chromatography, and radio-fluorescence high-performance liquid chromatography techniques applied to tissue extracts provided insight into the effects of BA treatment on ‘new’ carbon (as 11C) metabolism. Most notable was the strong influence reducing boron levels had on raising 11C partitioning into glutamine, aspartic acid, and asparagine. Altogether, the growth of maize under different regimes of boron affected 11CO2 fixation, its metabolism and allocation belowground, and altered root growth. Finally, inductively coupled plasma mass spectrometry provided insight into the effects of BA treatment on plant uptake of other essential nutrients. Here, levels of boron and zinc systematically increased in foliar tissues with increasing BA concentration. However, levels of magnesium, potassium, calcium, manganese, and iron remained unaffected by treatment. The rise in foliar zinc levels with increased BA concentration may contribute to improved 11CO2 fixation under these conditions.

Botanical description, bioactivity guided isolation and in silico mode of action of anti-diabetic constituents of Pterocarpus dalbergioides flowers

Diabetes mellitus, a serious disorder with associated life-threatening complications, needs effective preventive and therapeutic interventions. There is an utmost need for an effective, safe and economical anti-diabetic drug, thereby medicinal plants can be considered as a useful intervention. Pterocarpus dalbergioides, an evergreen tree native to the Andaman Island, has been employed as a folk remedy for diabetes. Botanical features of the flowers of P. dalbergioides were described by macroscopic and microscopic analysis of both the fresh flowers and their fine powder. Bioactivity-guided isolation focusing on the assessment of anti-diabetic activity of the flowers of the plant in alloxan-induced diabetic rats was performed. The molecular activity of three isolated compounds against six important targets having a prominent role in diabetes pathophysiology was also done. The results revealed that the n-butanol fraction prevented the diabetogenic effect of alloxan and decreased significantly the blood glucose level (p < 0.01) in treated rats. Three flavones, luteolin (F1), luteolin-7-O-glucoside (F2), and luteolin-7-O-rutinoside (F3) were isolated from the flower of the studied plant for the first time. A high-performance liquid chromatography technique (HPLC) was used to quantify luteolin as a major isolated phenolic compound. Docking-based mode of action prediction of isolated compounds revealed that all three compounds can act as an inhibitor of PTP1B, DPP4, α-amylase, and also an activator of PPAR-γ and FFAR1. Consequently, the current research includes botanical characteristics of the flowers of the plant which facilitate its authentication and indicates that P. dalbergioides flower could be of therapeutic benefit for the management of diabetes mellitus.

Chiral separation of calcium channel antagonists by SFC and HPLC using different immobilized chiral stationary phases.

Supercritical fluid chromatography and high-performance liquid chromatography techniques are popular for the chiral separations of different drugs and pharmaceuticals. Therefore, this article describes a comparative study of the chiral separation of some calcium channel antagonists such as verapamil, gallopamil, and nisoldipine. The columns used were Chiralpak IG and Chiralpak ID (250 mm × 4.6mm, 5.0μm). The separation was achieved by using a variety of mobile phases in both techniques. The retention, separation, and resolution factors in SFC were in the range of 1.36-7.30, 1.09-1.72, and 1.16-3.47, while these values in the case of HPLC were 1.03-2.42, 1.12-1.35, and 0.49-2.46. The complete resolution of gallopamil and verapamil was achieved successfully. The chiral recognition was controlled by hydrogen bondings, π-π interactions, dipole induced dipole interactions, van der Waal forces, and steric effects. SFC was found to be a better technique than HPLC because of quick separation, good separation power, economic, environment-friendly, and green technology.

Development and Validation of Analytical Test Method for the Determination of Oseltamivir Phosphate Residues on the Manufacturing Equipment Surfaces Equipment Using Swab and Rinse Sampling and by High Performance Liquid Chromatographic Technique

Development and Validation of Analytical Test Method for the Determination of Oseltamivir Phosphate Residues on the Manufacturing Equipment Surfaces Equipment Using Swab and Rinse Sampling and by High Performance Liquid Chromatographic Technique

High Performance Liquid Chromatographic Analysis and Antibacterial Activity of Methanolic Seed Extract of Syzygium cumini L.

Syzygium cumini is a medicinally important plant widely used in traditional systems of medicines specifically Ayurveda followed in India. The seeds of the plant are recognized in various bio medicines for the treatment of diabetes. It is reasonable source of natural antioxidants used in dietary supplement of food and bioactive substances in cosmetics and pharmaceutical industries. In the present study, Syzygium cumini (jamun) seeds were collected from the Alagar Kovil Hills and Procured from local Madurai central market of Madurai district, Tamil Nadu. The seed extract was analysed for checking the antibacterial activity and phytocomponents of seed extract by High Performance Liquid Chromatography technique. Different solvents like acetone, methanol, ethanol, butanol, petroleum ether and distilled water used for extracting the seed extract. The inhibition efficiency of seed extract was treated with bacterial pathogens namely: Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. Among these solvents, methanol was a suitable one for extracting the phytocompounds efficiently. The methanolic seed extract of Syzygium cumini analysed with high performance liquid chromatography (HPLC) and it reported that the presence of phenolic compounds specifically gallic acids in the crude extract. The presence of such compounds in Syzygium cumini seeds which confirmed that the positive effect of seed component could control the growth of bacterial pathogens under laboratory condition.

Evaluation of cardioprotective potential of isolated swerchirin against the isoproterenol-induced cardiotoxicity in wistar albino rats

Background: Cardiotoxicity is one of the emerging health-care issues worldwide and mortality due to this is increasing exponentially. Oxidative stress, myocardial inflammation, and damage to the cardiac membrane are major causative attributes for this toxicity. Isoproterenol (ISO)-induced cardiotoxicity is a well-established and accepted model to estimate innovative cardioprotective agents, as it exhibits several morphological and biochemical aberrations in the myocardium of rat that is comparable with those detected in clinical practice. Objectives: In the present study, we explored the cardioprotective effect of swerchirin (SW) extracted from Swertia chirayita in the ISO model. Materials and Methods: SW was isolated from S. chirayita, characterized by high-performance thin-layer chromatography-mass spectrometry, 1H-nuclear magnetic resonance (NMR), 13C-NMR, and high-performance liquid chromatography techniques. The in-vitro study was performed for 2,2-Diphenyl-1-picrylhydrazyl and nitric oxide scavenging activity to explore the antioxidant capacity. In silico study was performed to ascertain the binding affinity of SW with antioxidant enzymes. The in-vivo study was performed to explore the cardioprotective activity in the ISO-induced cardiotoxic model. Results: The in vitro studies showed the significant antioxidant potential of SW whereas the in silico study revealed its effective binding into the catalytic pocket domain of superoxide dismutase and catalase. The in vivo study showed a significant improvement in antioxidant enzymes and a reduction in serum glutamic-oxaloacetate transaminase and lipid profile. Further, SW also significantly reversed the histopathological aberrations in a dose-dependent manner. Conclusion: Findings of the study showed a significant cardioprotective effect of SW against ISO-induced cardiac damage.