Studies of Phytochemicals, Antioxidant, and Antibacterial Activities of Pinus gerardiana and Pinus roxburghii Seed Extracts.

Pine seeds are considered as nonwood forest products (NWFP) with regularly increasing market's demand. They can be eaten in various ways such as roasted or raw. In addition, they are included in various traditional dishes like in cookies, sauces, candies, cakes, breads, and other bakery items and, moreover, for medicinal purposes. GC-MS study is performed to analyze the phytochemical compounds present in the seed extracts of Pinus roxburghii (Chir) and Pinus gerardiana (Chilgoza). In total, 25 compounds were identified each in Chir and Chilgoza. In Chir seeds, abundantly present compounds were 2,4-di-tert-butylphenol (16.6%), followed by ç-Terpinene (9.9%) and cyclohexanol, 4-ethenyl-4-methyl-3-(1-methylethenyl)-, (1à,3à,4á) (9.8%), whereas in Chilgoza seeds, the maximum amount of compound was 1-hexyl-1-nitrocyclohexane (17.3%), followed by phenol, 2,6-bis(1,1-dimethylethyl) (15.4%), and heptadecane, 2-methyl (8.4%). The total phenolic content of Chir seed sample was 1536 ± 4.35 (mg GAE/100 g), whereas in the Chilgoza seed extract was 642.66 ± 2.08 (mg GAE/100 g). The application of RP-HPLC-DAD system revealed that Chir and Chilgoza seeds have maximum quantity of catechin (15.77 ± 0.16 µg/mg and 17.49 ± 0.32 µg/mg, respectively). Both Chir and Chilgoza seed extracts exhibited significant antioxidant (radical scavenging) potential, through H2O2 (618.94 ± 21.45 µg/mL and 575.16 ± 19.88 µg/mL) and DPPH (552.60 ± 13.03 µg/mL and 429.15 ± 3.80 µg/mL) assays, respectively. Additionally, a well-known antibacterial potential was also found in both plants' dichloromethane extracts, with 64 to 256 µg/mL of minimum inhibitory concentrations. As a whole, result shows the importance of both plants as a naturally occurring phytochemical source with significant antibacterial and antioxidant activity.

Conifers Phytochemicals: A Valuable Forest with Therapeutic Potential.

Conifers have long been recognized for their therapeutic potential in different disorders. Alkaloids, terpenes and polyphenols are the most abundant naturally occurring phytochemicals in these plants. Here, we provide an overview of the phytochemistry and related commercial products obtained from conifers. The pharmacological actions of different phytochemicals present in conifers against bacterial and fungal infections, cancer, diabetes and cardiovascular diseases are also reviewed. Data obtained from experimental and clinical studies performed to date clearly underline that such compounds exert promising antioxidant effects, being able to inhibit cell damage, cancer growth, inflammation and the onset of neurodegenerative diseases. Therefore, an attempt has been made with the intent to highlight the importance of conifer-derived extracts for pharmacological purposes, with the support of relevant in vitro and in vivo experimental data. In short, this review comprehends the information published to date related to conifers' phytochemicals and illustrates their potential role as drugs.

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.

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 detection.

Bioassay guided fractionation of rhizome extract of Rheum emodi wall as bio-availability enhancer of antibiotics against bacterial and fungal pathogens.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheum emodi Wall., is an important medicinal plant extensively used in Ayurvedic and Unani systems of traditional medicine. It is known to possess antioxidant, antibacterial, antifungal, anticancer, wound healing and immune enhancing activities. AIM OF THE STUDY: The aim of the current study was to investigate the antimicrobial activity and synergistic potential of different solvent fractions and phytocompounds of Rheum emodi rhizome against bacterial and fungal pathogens. MATERIAL AND METHODS: The antimicrobial and synergistic potential of the crude methanolic extract, different solvent fractions (n-hexane, chloroform, ethyl acetate, and residual aqueous) and isolated phytocompounds of the rhizome of Rheum emodi were assayed by broth microdilution method. The bioactive phytocompounds were isolated through silica TLC and quantified using HPTLC and HPLC. The bioactive phytocompounds were identified by LC-MS analysis. RESULTS: Phytochemical analysis of the sub-fractions showed that the TPC (417.94 ± 1.2 mg g-1 GAE) and TFC (187.40 ± 0.5 mg g-1 RE) were highest in residual aqueous extracts. The chloroform sub-fraction possessed the highest antimicrobial activity against bacterial (Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae) and fungal strains (Candida albicans MTCC 277 and ATCC 90028). The MIC of chloroform sub-fraction against S. aureus, K. pneumoniae, E. coli, C. albicans was 1.95, 3.91, 15.62 and 62.5 µg ml-1, respectively. TLC and LC-MS analysis of chloroform sub-fraction identified phytocompounds namely emodin D4 (m/z 274.262), rhein13c6 (m/z 290.176), chrysophanol dimethyl ether (m/z 282.291), and resveratrol (m/z 340.456). Quantification of emodin content showed that the chloroform sub-fraction (101.4543 µg mg-1, 194.8037 µg mg-1 measured through HPTLC and HPLC, respectively), and its TLC fraction (II) (75.18 µg mg-1, 232.384 µg mg-1 measured through HPTLC and HPLC, respectively) are rich in emodin. Furthermore, chloroform sub-fraction, its TLC fractions and emodin showed profound synergistic activity in combination with antibacterial and antifungal antibiotics and lowered the dosage of antibiotics by 4-257 folds. CONCLUSIONS: The bioassay guided fractionation of R. emodi rhizome methanolic extract identified phytocompounds (emodin, rhein13c6, chrysophanol dimethyl ether and resveratrol) that act as bioavailability enhancers of antibacterial and antifungal antibiotics, hence revealing their potential in treating multidrug resistance.

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

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 ß-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 ß-sitosterol constituents of the plant.