NOVEL RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF SAXAGLIPTIN AND GLIMEPIRIDE

Objective: The objective of the present study was to develop, and validate a novel reverse phase high performance liquid chromatographic (RP-HPLC) method, for simultaneous determination of saxagliptin (SAXA) and glimepiride (GLIM), in bulk mixtures, and in tablets.Methods: Determination of the drugs, SAXA and GLIM, was carried out employing ODS C18 column (250 mm X 4.6 mm i. d, 5 µm particle size), with diode array detector at λmax of 230 nm. The mobile phase employed for the current study, composed of two solvents, i.e., A (acetonitrile), and B (0.1 % w/v sodium di-hydrogen orthophosphate buffer, pH 3.8 adjusted with orthophosphoric acid). The mobile phase was pumped at a flow rate of 0.75 ml/min in the gradient mode. The validation study with respect to specificity, linearity, precision, accuracy, robustness, limit of detection (LOD), and limit of quantification (LOQ), was carried out employing the ICH Guidelines.Results: The developed method was selective and linear for both the drugs, i.e., between 15.63 µg/ml and 250.00 µg/ml for SAXA, and 7.81 µg/ml and 125.00 µg/ml for GLIM, with a correlation coefficient (R2) 0.9977 and 0.9982, for SAXA, and GLIM, respectively. The % recovery obtained was 102.98±0.14% for SAXA, and 101.84±1.96% for GLIM. The LOD and LOQ values for SAXA were obtained to be 1.30 µg/ml, and 3.94 µg/ml, respectively, while for GLIM, LOD was 0.82 µg/ml and LOQ was 2.48 µg/ml. The method also exhibits good robustness for different chromatographic conditions like wavelength, flow rate, mobile phase and injection volume.Conclusion: The method was successfully employed, for the quantification of SAXA and GLIM, in the quality control of in-house developed tablets, and can be applied for the industrial use

Acid Rain and Seed Germination: A Predictive Model Using ML-based CART Algorithm

The impact of acid rain on the germination of seeds is a significant concern in agricultural and environmental studies. Acid rain, characterized by elevated acidity levels due to pollutants like sulfur dioxide and nitrogen oxides, can adversely affect the germination process of various plant species. The objective of this study was to evaluate the impact of simulated acid rain (SAR) on the germination of Brinjal (Solanum melongena Linn.) and Cowpea (Vigna unguiculata ssp. cylindrica L. Walpers) crops. The experiments were conducted using eight plastic trays of approximately 25 cm. x 30 cm dimensions. Four trays were used for experiments with brinjal seeds (Set I), while the other four were used for cowpea seeds (Set II). One tray from each set was used as positive control and treated with normal pH 5.6, while the other three trays from each batch were treated with SAR solutions of pH 4.5, 3.5, and 2.5. Brinjal seed germination percentage and seed vigor were inferior to Cowpea seeds. The seeds treated with SAR (pH 4.5, 3.5, and 2.5) showed hindered seed germination. Furthermore, a more significant inhibitory effect was observed at lower pH values. The mean germination percentage of seeds was highest for standard SAR (pH 5.6) in the case of Brinjal seeds, while it was recorded lowest for Cowpea seeds. The results indicate that plants do not respond uniformly to SAR. To investigate the behavior of the simulated acid rain data, a Machine Learning-based Decision Tree Algorithm was employed to identify and optimize conditions. Cowpea was predicted to get 95% seed germination, whereas brinjal would only be 64% in acid rain of pH value 5.05 for 36 hours. In conclusion, utilizing a Machine Learning-based CART algorithm has provided valuable insights into predicting the germination behavior of seeds under the influence of acid rain

Understanding of colistin usage in food animals and available detection techniques: a review

Progress in the medical profession is determined by the achievements and effectiveness of new antibiotics in the treatment of microbial infections. However, the development of multiple-drug resistance in numerous bacteria, especially Gram-negative bacteria, has limited the treatment options. Due to this resistance, the resurgence of cyclic polypeptide drugs like colistin remains the only option. The drug, colistin, is a well-known growth inhibitor of Gram-negative bacteria like Acinetobacter baumanni, Enterobacter cloacae, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Technological advancements have uncovered the role of the mcr-1(mobilized colistin resistance) gene, which is responsible for the development of resistance in Gram-negative bacteria, which make them distinct from other bacteria without this gene. Additionally, food animals have been determined to be the reservoir for colistin resistance microbes, from which they spread to other hosts. Due to the adverse effects of colistin, many developed countries have prohibited its usage in animal foods, but developing countries are still using colistin in animal food production, thereby imposing a major risk to the public health. Therefore, there is a need for implementation of sustainable measures in livestock farms to prevent microbial infection. This review highlights the negative effects (increased resistance) of colistin consumption and emphasizes the different approaches used for detecting colistin in animal-based foods as well as the challenges associated with its detectio

Azelaic acid and Melaleuca alternifolia essential oil co-loaded vesicular carrier for combinational therapy of acne

Aim: Azelaic acid (AzA), a comedolytic, antibacterial, anti-inflammatory anti-melanogenic agent, prescribed against acne vulgaris is safe on skin. Its combination with another widely used anti-acne agent, tea tree oil (EO) whose delivery is limited by volatility, instability and lipophilicity constraints was attempted. Method: Solvent injection was used to prepare AzA-EO integrated ethosomes. Result: Ethosomes were transformed into carbopol hydrogel, which exhibited pseudo-plastic properties with appreciable firmness, work of shear, stickiness and work of adhesion. The hydrogel showed better permeation and retention characteristics vis-a-vis commercial formulation (AzidermTM), when evaluated in Wistar rat skin. Further, ethosome hydrogel composite was better tolerated with no side effects. Conclusion: The findings suggests that the aforementioned strategy could be a potential treatment used for acne management. </jats:p

Hydrogel composite containing azelaic acid and tea tree essential oil as a therapeutic strategy for Propionibacterium and testosterone-induced acne.

Azelaic acid (AzA) is a USFDA bioactive prescribed against acne vulgaris. It possesses delivery challenges like poor aqueous solubility, low skin-penetrability, and dose-dependent side effects, which could be overcome by its synergistic combination with tea tree oil (TTO) as a microemulsion (ME)-based hydrogel composite. AzA-TTO ME was prepared to employ pseudo-ternary phase diagram construction. The best AzA-TTO ME was of uniform size (polydispersity index  90%), and negative zeta potential (-1.42 ± 0.25% mV) values. ME hydrogel composite with optimum rheological and textural attributes showed better permeation, retention, and skin-compliant characteristics, vis-a-vis marketed formulation (Aziderm™) when evaluated in Wistar rat skin. In vitro antibacterial efficacy in bacterial strains, i.e., Staphylococcus aureus, Propionibacterium acne, and Staphylococcus epidermidis, was evaluated employing agar well plate diffusion and broth dilution assay. ME hydrogel has shown an increase in zone of inhibition by two folds and a decrease in minimum inhibitory concentration (MIC) by eightfold against P. acnes vis-a-vis AzA. Finally, ME hydrogel composite exhibited a better reduction in the papule density (93.75 ± 1.64%) in comparison to Aziderm™ 72.69 ± 4.67%) on acne as developed in rats by inducing testosterone. Thus, the developed AzA-TTO ME hydrogel composite promises an efficacious and comparatively safer drug delivery system for the topical therapy of acne vulgaris

Laparoscopic Versus Open Sigmoid Loop Colostomy: A Comparative Study from a Cohort of 62 Patients Requiring Temporary Faecal Diversion at a Tertiary Care Center in North India

Background:&nbsp;Over decades, colostomies have been done through open method, but laparoscopic creation of an intestinal stoma is safe, feasible and has distinct advantages over conventional techniques in specific procedures. The aim of this study compares operative and short-term outcomes of laparoscopic and open sigmoid loop colostomy formation for temporary fecal diversion.&nbsp;Subjects and Methods:&nbsp;A single institution, comparative study conducted in the department of surgery for patients who underwent either laparoscopic or open sigmoid loop colostomy. The 2 years' study was from December 1, 2013, to November 30, 2015. Subjects were prospectively enrolled in the study after informed consent, both genders of &gt;12 years of age. Data analysis was done using Statistical Package for Social Sciences version 21.0. Variables were tested by Kolmogorov–Smirnov test, compared using unpaired&nbsp;t-test/Mann–Whitney Test, Chi-square test/Fisher's exact test.&nbsp;P&nbsp;&lt; 0.05 was considered statistically significant.&nbsp;Results:&nbsp;Sixty-two patients were enrolled; laparoscopy group – 29 patients (46.77%) versus open group – 33 patients (53.22%). Laparoscopic group/open surgery group showed less blood loss&nbsp;(20.69 + 17.71 ml / 121.97 + 35.29ml,&nbsp;P-value 0.0005), lower requirement of analgesics (4.28 ± 1.76 days/6.88 ± 2.75 days), shorter hospital stay (8.79 ± 5.57 days and 11.73 ± 6.61 days,&nbsp;P = 0.001), early return of the bowel function and tolerance to diet. Complications and readmission requirement for any complication was lower in the laparoscopic group. Conclusions:&nbsp;Laparoscopic sigmoid loop colostomy is a simple alternative to open sigmoid loop colostomy with respect to postoperative pain, earlier return of bowel function, lower analgesic requirement, and lesser hospital stay

In-vitro osteoblast proliferation and in-vivo anti-osteoporotic activity of Bombax ceiba with quantification of Lupeol, gallic acid and β-sitosterol by HPTLC and HPLC

Abstract Background Bombax ceiba is used traditionally to treat bone disorders, rheumatism, and joint pain. The aim of the study is to carry out osteogenic activity in-vitro and anti-osteoporotic activity in-vivo of stem bark of B. ceiba in surgical ovariectomy model in female rats. Methods Plant drug: B. ceiba stem bark was extracted with solvents petroleum ether and methanol using Soxhlet extraction. In-vitro osteoblastic proliferation study was performed using UMR-106 cell lines. Both the extracts were undergone to acute toxicity study as per OECD423 guidelines. Female Wistar albino rats 180-240 g were used (n = 6). Surgical ovariectomy was performed under anesthesia to induce bone porosity and loss in all animals except normal control and sham control. Each extract was administered at two dose level: 100 and 200 mg/kg and the standard Raloxifene was given at 1 mg/kg orally for 28 days. The phytochemical study of both the extracts was performed using HPLC and HPTLC. Results A significant osteoblast cell proliferation and alkaline phosphatase activity were observed with B. ceiba extracts in UMR-106 cell lines. Surgical removal of ovaries produced significant (p < 0.05) decline in bone mineral density, bone breaking strength, serum ALP, calcium, phosphorus, and estradiol level and marked bone tissue destruction in histology. Administration of petroleum ether and methanolic extract for 28 days significantly (p < 0.05) ameliorated the consequences of ovariectomy induced bone porosity and restored the normal architecture of bone, as compared to OVX control. The phytochemical screening of both the extracts were also carried out. The quantification of phytoconstituents showed the presence of β-sitosterol and lupeol in petroleum ether extract, whereas the lupeol is also quantified in the methanolic extract. The presence of gallic acid was quantified in methanolic extract using HPLC. Conclusion B. ceiba: stem bark ameliorated the state of bone fragility and fracture possibly due to estrogenic modulation, as also confirmed by in-vitro osteogenic activity which may be due to the presence of lupeol, gallic acid and β-sitosterol constituents of the plant

Determination of Colistin B in Chicken Muscle and Egg Using Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Colistin, an imperative member of the polymyxin group, is a cationic peptide antibiotic. Itis also known as polymyxin E, but this peptide antibiotic has been forbidden for human consumption due to its high toxicity. Regrettably, this antibiotic is utilized as a feed additive and veterinary drug for animals. Due to the toxicity of colistin, the presence of its residue in the animal system represents a threat to human health regarding the consumption of meat, especially chicken. A novel method was proposed for quantifying colistin B in chicken muscles and eggs using ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS). In this method, extraction of colistin B from samples was achieved by mixing the sample with acidified methanol:water (1/1, v/v), followed by centrifugation and filtration by a membrane filter excluding solid-phase extraction (SPE) clean up, as well as evaporation steps. The analysis was conducted by optimized liquid chromatography–tandem mass spectrometry (LC–MS/MS), and method performance was assessed in terms of the limit of quantitation, specificity, selectivity, precision, linearity and recovery in coherence with the guidelines of SANTE and the Commission Decision 2002/657/EC. The result obtained from the study showed the limit of quantitation (LOQ) as 10 µg Kg−1 for muscles and 5 µg Kg−1 for eggs, with acceptable recoveries along with precision. The linearity was plotted in the range of 5–25 µg L−1 (solvent) for egg and 10–50 µg Kg−1 (matrix-matched) for muscles. The result of average recoveries showed the value of 70–94% (3.3–12% relative standard deviation (RSD)) for chicken muscles and 88–107% (2.5–18.6% RSD) for egg samples, which meets the criteria for acceptability of method according to both SANTE and 2002/657/EC guidelines. This proposed protocol provides a cost-effective solution for food testing labs by reducing the cost of the sample preparation by 60% along with the time required for SPE cleanup. Further, the optimized method was also tested on real samples collected from nearby provinces in Solan city, Himachal Pradesh, India, and three out of 20 muscles were found to have colistin B in the range of 50–560 µg Kg−1

Gastric ulcer healing by chebulinic acid solid dispersion-loaded gastroretentive raft systems: preclinical evidence

Background: Chebulinic acid (CA), a component in Terminalia chebula, exhibits antiulcer activity, but has poor aqueous solubility. Raft-forming systems incorporating solid dispersions (SDs) of CA, were developed to overcome its poor biopharmaceutical properties and to prolong the gastric residence time for maximum activity. Methods: SDs were formulated by a solvent evaporation method using Eudragit EPO. Raft formulations consisted of sodium alginate as a polymer. Results: Release of CA in the dissolution medium was 40%, whereas SDs showed 95.45% release. The CA raft system (20 mg/kg) showed curative efficacy in an alcohol-induced gastric ulcer model and increased protection when compared with omeprazole (10 mg/kg) and CA suspension (20 mg/kg). Conclusion: These studies demonstrated SD raft systems to be a promising approach for antiulcer therapy by CA. </jats:p

Phytochemistry, Pharmacology, and Nutraceutical Profile of <i>Carissa</i> Species: An Updated Review

Carissa, a genus of the Apocynaceae family, consists of evergreen species, such as shrubs as well as small trees that are native to Asia, Africa, and Oceania’s subtropical and tropical regions. Most of the Carissa species are traditionally used to treat various diseases, such as chest pain, headaches, gonorrhoea, rheumatism, syphilis, oedema, rabies, stomach pain, hepatitis, cardiac diseases, and asthma. The pharmacological studies on Carissa species revealed its antioxidant, antimicrobial, anticancer, cardioprotective, antipyretic, analgesic, wound healing, anticonvulsant, antiarthritic, adaptogenic, anti-inflammatory, and antidiabetic activities, thus validating its use in indigenous medicine systems. The review article summarised the comprehensive literature available, including morphology, indigenous uses, bioactive composition, nutraceutical, and pharmacological activities of Carissa species. A total of 155 research papers were cited in this review article. The Carissa fruits are rich in dietary fibre, lipids, proteins, carbohydrates, vitamin C, and macro- and micro-elements. A total of 121 compounds (35 polyphenols (flavonoids and phenolic acids), 30 lignans, 41 terpenoids, 7 steroids, 2 coumarins, and 6 cardiac glycosides) have been extracted from C. spinarum, C. carandas, and C. macrocarpa. Among all chemical constituents, lupeol, carissol, naringin, carisssone, scopoletin, carissaeduloside A, D, J, carandinol, sarhamnoloside, carissanol, olivil, carinol, 3β-hydroxyolean-11-en-28,13β-oilde, ursolic acid, and carissone are the key bioactive constituents responsible for pharmacological activities of genus Carissa. The gathered ethnopharmacological information in the review will help to understand the therapeutic relevance of Carissa as well as paving a way for further exploration in the discovery of novel plant-based drugs