Improved up-and-down procedure for acute toxicity measurement with reliable LD50 verified by typical toxic alkaloids and modified Karber method

Background Up-and-down procedure (UDP) was recommended to replace traditional acute toxicity methods. However, it was limited due to the long experimental period (20–42 days). To improve UDP, an improved UDP method (iUDP) was developed by shortening observation time between sequence dosages. The aim of this study was to test the reliability of iUDP to provide a reliable method for the acute toxicity measurement of valuable or minor amount compounds. Methods Oral median lethal dose (LD50) of nicotine, sinomenine hydrochloride and berberine hydrochloride were measured both by iUDP and modified Karber method (mKM). Results LD50 of the three alkaloids measured by iUDP with 23 mice were 32.71 ± 7.46, 453.54 ± 104.59, 2954.93 ± 794.88 mg/kg, respectively. LD50 of the three alkaloids measured by mKM with 240 mice were 22.99 ± 3.01, 456.56 ± 53.38, 2825.53 ± 1212.92 mg/kg, respectively. The average time consumed by the two methods were 22 days and 14 days respectively. Total grams of the alkaloids used by the two methods were 0.0082 and 0.0673 (nicotine), 0.114 and 1.24 (sinomenine hydrochloride), 1.9 and 12.7 (berberine hydrochloride). Conclusion iUDP could replace mKM to detect acute toxicity of substances with comparable and reliable result. And it is suitable for valuable or minor amount substances.

Microwave Assisted Extraction of Berberine and Preparation of Berberine Hydrochloride from Berberis Aristata Variety of Nepal, and Quantification using RP-HPLC and HPTLC Methods

Berberis aristata a Himalayan woody spiny shrub with yellow flowers and red berries commonly called as Daruharidra in Sanskrit and locally in Nepal is called as Chutro or Chitra. The root and stem are the two parts widely used in traditional medicines of India and China. Berberine is the key active ingredient present in stem and root parts. Berberine hydrochloride is the derivative of berberine. The present study aimed to study the microwave assisted extraction of berberine and its conversion into berberine hydrochloride and quantifying by RP-HPLC and HPTLC methods.In the present paper we have mentioned microwave assisted extraction of berberine and preparation of berberine hydrochloride in detail. Berberine extracted from roots of Berberis aristata using microwave assisted extraction in 80% ethanol to obtain 20% pure berberine crude by HPTLC densitometry at 350 nm absorption. The crude berberine was further purified to berberine hydrochloride by adding 10% Hcl in aqueous solution of berberine and allowed to crystallize at 5 0C over 24 hours. The crystals were further purified and recrystallized in ethanol and subjected to RP-HPLC. Reverse phase HPLC was carried out on Shimadzu UV detector at wave length of 265 nm using Acetonitrile-0.1% phosphoric acid solution (50:50) (add 0.1g sodium dodecyl sulfonate per 100ml) as the mobile phase; Phenomenex RP-column (250 mm × 4.6 mm, 5 μm), With flow rate 1.0 mL/minute, 5 µL injection volume, column temperature 25 0C for run time of 35 minutes, and retention time of berberine hydrochloride was 12.008 minutes with purity of 82% and recovery of 90% yield obtained.

Molecular Characterization of Prdx Family and Response of Antioxidant Enzymes in Berberine Hydrochloride-Treated Charybdis japonica Infected With Aeromonas hydrophila

Berberine hydrochloride is an isoquinoline alkaloid, which has antitumoral, antibacterial, and antiviral activities in vivo and in vitro. Charybdis japonica is one of the main economic species of crab in Southeast Asia. We studied the molecular mechanism of oxidative stress in berberine hydrochloride-treated C. japonica infected with Aeromonas hydrophila. C. japonica were infected with A. hydrophila after being submerged in different concentrations (0, 100, 200, and 300 mg/L) of berberine hydrochloride for 48 h. The full-length cDNA of Prx6 and the ORFs of Prx5 and PXL2A were cloned. Prx6 and PXL2A each have one conserved domain, Cys44, and Cys81. The Prx5 conserved domain contains three important Cys loci, Cys75, Cys100, and Cys76. Prx6 was different from Prx5 and PXL2A in the Peroxiredoxin family. The transcription levels of PXL2A infected with A. hydrophila were all higher than the control. The transcription levels of C. japonica were further increased by adding berberine hydrochloride and were increased the highest at a concentration of 300 mg/L. The activities of glutathione peroxidase, superoxide dismutase, and catalase in the hepatopancreas of berberine hydrochloride-treated C. japonica infected with A. hydrophila were significantly increased compared with those only infected with A. hydrophila and the control group. The glutathione transferase activity in the hepatopancreas was significantly increased in berberine hydrochloride-treated C. japonica. The results of this study provide a new understanding of the potential role of berberine hydrochloride on the oxidative stress mechanisms of C. japonica.

Effect of Berberine Hydrochloride Treatment on Transcriptomic Response in Gill Tissue of Charybdis japonica Infected With Aeromonas hydrophila

Berberine hydrochloride is the main active ingredient of Coptis chinensis, which has demonstrated significant antibacterial activity against bacteria. However, the molecular mechanism underlying its effects is unclear in crabs. In this study, we used transcriptome sequencing to investigate the molecules involved in the therapeutic and defensive responses of Charybdis japonica infected with 105 colony-forming units (CFU)/L of Aeromonas hydrophila following treatment with berberine hydrochloride. A total of 5,409 differentially expressed genes were observed after exposure of C. japonicus for 72 h to 100 mg/L berberine hydrochloride, 100 mg/L berberine hydrochloride plus injection of 105 CFU/L A. hydrophila, and a control group injected with equal amount of physiological saline solution. Enrichment analysis revealed that these genes were involved in metabolism, cellular processes, signal transduction, and immune function. The transcriptomic results indicated that exposure to berberine hydrochloride activated glutathione metabolism, oxidative phosphorylation, fatty acid degradation, retinol metabolism, pyruvate metabolism, and the hypoxia-inducible factor 1 signaling pathway. Transcriptomic analysis and relative mRNA level analysis also identified acid hydrolase genes and phagosomal genes as key factors regulating the adaptation of acid berberine-infused C. japonicus to infection with A. hydrophila, Based on the ability of berberine hydrochloride to induce acid hydrolase and phagosomal gene expression, which can in turn remove bacteria, immersion in berberine hydrochloride may remove A. hydrophila and thus improve the survival of C. japonica. The results of this study provided a new scientific basis for the potential role of berberine hydrochloride in the immune mechanisms of crabs.

Rice husk hydrochars from metal chloride-assisted hydrothermal carbonization as biosorbents of organics from aqueous solution

Hydrochar a carbon-rich material resulting from hydrothermal carbonization of biomass, has received substantial attention because of its potential application in various areas such as carbon sequestration, bioenergy production and environmental amelioration. A series of hydrochars were prepared by metal chloride-assisted hydrothermal carbonization of rice husk and characterized by elemental analysis, zeta potential, X-ray diffraction, Brunauer–Emmett–Teller measurements, Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray photoelectron spectroscopy and scanning electron microscopy. The results reveal that the prepared hydrochars have carbon contents ranging from 45.01 to 58.71%, BET specific areas between 13.23 and 45.97 m2/g, and rich O-containing functional groups on the surfaces. The metal chlorides added in the feedwater could improve the degree of carbonization and show significant effects on the physical, chemical and adsorption properties of the hydrochars. The adsorption of the selected organics on the hydrochars is a spontaneous and physisorption-dominated process. The hydrochars possess larger adsorption capacities for 2-naphthol than for berberine hydrochloride and Congo red, and the modeling maximum adsorption capacities of 2-naphthol are in the range of 170.1–2680 mg/g. The adsorption equilibrium could be accomplished in 10, 40 and 30 min for 2-naphthol, berberine hydrochloride and Congo red, respectively. These results suggest metal chloride-assisted hydrothermal carbonization a promising method for converting biomass waste into effective adsorbents for wastewater treatment.