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      Policosanol fabrication from insect wax and optimization by response surface methodology

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          Abstract

          Background

          Insect wax is a famous biological resource for the role in economic production in China. Insect wax is a good source of policosanol, which may is a candidate supplement in foodstuff and pharmaceuticals that has important physiological activities. Therefore, this work aims to investigate a high-yield and rapid method for policosanol fabrication from insect wax.

          Results

          The conditions for policosanol fabrication were optimized as follows: an oil bath temperature of 112.7°C and reductant dosage of 0.97 g (used for the reduction of 10.00 g of insect wax). The yield of policosanol reached 83.20%, which was 4 times greater than that of existing methods, such as saponification. The total content of policosanol obtained under the optimal conditions reached 87%. In other words, a high yield of policosanol was obtained from insect wax (723.84 mg/g), that was 55 times higher than that generated from beeswax-brown via saponification. The concentrations of metal residues in policosanol were within the limits of the European Union regulations and EFSA stipulation. The LD50 values for oral doses of insect wax and policosanol were both > 5 g/kg.

          Conclusion

          Policosanol was fabricated via solvent-free reduction from insect wax using LiAlH 4 at a high yield. The fabrication conditions were optimized. Policosanol and insect wax showed high security, which made them potential candidates as supplements in foods, pharmaceuticals and cosmetics. The rapid and high-yield method has great potential for commercial manufacturing of policosanol.

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          Most cited references29

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          Chitosan-sodium alginate nanoparticles as submicroscopic reservoirs for ocular delivery: formulation, optimisation and in vitro characterisation.

          Management of extraocular disease is mainly limited by the inability to provide long-term extraocular drug delivery without avoiding the systemic drug exposure and/or affecting the intraocular structures and poor availability of drugs, which may be overcome by prolonging the contact time with the ocular surface, for instance with bioadhesive polymers. In the present study, mucoadhesive chitosan (CS)-sodium alginate (ALG) nanoparticles were investigated as a new vehicle for the prolonged topical ophthalmic delivery of antibiotic, gatifloxacin. A modified coacervation or ionotropic gelation method was used to produce gatifloxacin-loaded submicroscopic nanoreservoir systems. It was optimised using design of experiments by employing a 3-factor, 3-level Box-Behnken statistical design. Independent variables studied were the amount of the bioadhesive polymers: CS, ALG and the amount of drug in the formulation. The dependent variables were the particle size, zetapotential, encapsulation efficiency and burst release. Response surface plots were drawn, statistical validity of the polynomials was established and optimised formulations were selected by feasibility and grid search. Nanoparticles were characterised by FT-IR, DSC, TEM and atomic force microscopy. Drug content, encapsulation efficiency and particle properties such as size, size distribution (polydispersity index) and zetapotential were determined. The designed nanoparticles have average particle size from 205 to 572 nm (polydispersity from 0.325 to 0.489) and zetapotential from 17.6 to 47.8 mV. Nanoparticles revealed a fast release during the first hour followed by a more gradual drug release during a 24-h period following a non-Fickian diffusion process. Box-Behnken experimental design thus facilitated the optimisation of mucoadhesive nanoparticulate carrier systems for prolonged ocular delivery of the drug.
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            Comparison of the up-and-down, conventional LD50, and fixed-dose acute toxicity procedures.

            The up-and-down procedure (UDP), fixed-dose procedure (FDP) and conventional LD50 tests were compared to determine their consistency in chemical hazard classification for acute oral toxicity according to the European Economic Community (EEC) system. There was consistent classification for 23 out of 25 cases between the UDP and the conventional LD50 results, in 16 out of 20 cases between the FDP and the conventional LD50, and in seven out of 10 cases between the UDP and the FDP. The UDP needed only between six and 10 animals of one sex (fewer than either the LD50 or the FDP). Available literature indicates that the sexes are usually similar in their acute toxicity responses and that of females are often more sensitive than males when acute toxicity differences do exist, thus obviating the need for both sexes to be tested in most cases. Unlike the FDP, the UDP also estimates an LD50, thus providing data directly applicable to all current hazard classification systems based on acute oral toxicity.
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              Acute toxicity and sub-chronic toxicity of steroidal saponins from Dioscorea zingiberensis C.H.Wright in rodents.

              Steroidal saponins from Dioscorea zingiberensis are widely used in China for curing cardiovascular diseases. However, there was little toxicological information available on them. The study evaluated potential toxicity of the steroidal saponins and analyzed the metabolites in rats. For the acute study, the steroidal saponins were administered to kunming mice in single doses of 112.5-9000 mg/kg given by gavage. General behavior adverse effects, mortality and liver histopathological changes were examined. For the sub-chronic toxicity study, Sprague-Dawley rats were gavaged at doses of 127.5, 255 and 510 mg/kg/day for 30 days, then examined the biochemical and hematological parameters. Metabolites in serum were analyzed by HPLC-MS. The steroidal saponins caused dose-dependent general behavior adverse effects, mortality and liver histopathological changes in the acute toxicity study. In the sub-chronic toxicity study, 510 mg/kg/day of steroidal saponins increased total bilirubin (TBIL) in serum and decreased protein content in liver significantly. The metabolic process of TBIL in liver includes TBIL intaking, conjugated bilirubin forming, conjugated bilirubin excreting to biliary passage. Treatment with high dose of the steroidal saponins in vivo may lead to vacuolization of the cytoplasm of hepatocytes and canalicular cholestasis. In all doses used in the experiment, the steroidal saponins decreased aspartate aminotransferase (GOT), alanine aminotransferase (ALT) and alkaline phosphatase (AKP) in serum and increased reduced glutathione hormone (GSH), glutathione reductase (GR) and glutathione S-transferases (GST) in liver. Diosgenin was the main metabolite in serum. The steroidal saponins did not show any sign of toxicity up to oral dose of 562.5mg/kg in mice. No significant changes of biochemical and hematological parameters in rats (except at 510 mg/kg/day), it was concluded that the steroidal saponins did not appear to have significant toxicity in their traditional uses.
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                Author and article information

                Contributors
                Role: Formal analysisRole: InvestigationRole: VisualizationRole: Writing – original draft
                Role: Data curationRole: Project administrationRole: Validation
                Role: Methodology
                Role: Writing – review & editing
                Role: Software
                Role: Writing – review & editing
                Role: Funding acquisition
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                15 May 2018
                2018
                : 13
                : 5
                : e0197343
                Affiliations
                [001]Research Institute of Resources Insects, Chinese Academy of Forestry, Kunming, Yunnan, China
                College of Agricultural Sciences, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0001-6543-4449
                Article
                PONE-D-17-36630
                10.1371/journal.pone.0197343
                5953464
                29763430
                9d172760-afe8-4f32-8bb0-0db0ba361f56
                © 2018 Ma et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 12 October 2017
                : 21 April 2018
                Page count
                Figures: 5, Tables: 3, Pages: 14
                Funding
                Funded by: the Fundamental Research Funds for the Central Non-profit Research Institution of CAF
                Award ID: CAFYBB2018SY025
                Award Recipient :
                Funded by: National High Technology Research and Development Program of China (863 Program)
                Award ID: 2014AA021801
                Award Recipient :
                This work was financially supported by the Fundamental Research Funds for the Central Non-profit Research Institution of CAF (CAFYBB2018SY025) http://www.caf.ac.cn/, and National High Technology Research and Development Program of China (863 Program) (2014AA021801) http://program.most.gov.cn/.
                Categories
                Research Article
                Physical Sciences
                Materials Science
                Organic Materials
                Waxes
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Invertebrates
                Arthropoda
                Insects
                Biology and Life Sciences
                Biochemistry
                Lipids
                Oils
                Research and Analysis Methods
                Separation Processes
                Fractionation
                Organic Fractionation
                Saponification
                Biology and Life Sciences
                Zoology
                Entomology
                Insect Physiology
                Biology and Life Sciences
                Zoology
                Animal Physiology
                Invertebrate Physiology
                Insect Physiology
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Grasses
                Rice
                Research and Analysis Methods
                Experimental Organism Systems
                Plant and Algal Models
                Rice
                Biology and Life Sciences
                Ecology
                Plant Ecology
                Plant-Animal Interactions
                Plant-Insect Interactions
                Ecology and Environmental Sciences
                Ecology
                Plant Ecology
                Plant-Animal Interactions
                Plant-Insect Interactions
                Biology and Life Sciences
                Plant Science
                Plant Ecology
                Plant-Animal Interactions
                Plant-Insect Interactions
                Physical Sciences
                Chemistry
                Chemical Compounds
                Organic Compounds
                Chloroform
                Physical Sciences
                Chemistry
                Organic Chemistry
                Organic Compounds
                Chloroform
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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