Statistical mixture design investigation of fractionated and total extracts from Erythrina speciosa Andrews leaves

The simplex centroid mixture design for the ethanol, dichloromethane, hexane and acetone solvents has been applied to the extraction of crude mass and the fiber, organic, neutral and basic fractions as well as the fractionation residues of Erythrina speciosa Andrews leaves. Binary and ternary synergic solvent interactions are seen to provide dominant contributions to the extraction of both crude mass and all the fractions. Quadratic and special cubic mixture models precisely predict the extracted quantities of each fraction and the residue as a function of the proportions of the four solvents. Different solvent mixtures are found to be the most efficient extractors for the different fractions: binary dichloromethane‐hexane mixtures for the fiber fraction, ternary ethanol‐dichloromethane‐acetone mixtures for the neutral fraction, binary ethanol‐dichloromethane mixtures for the organic fraction, crude extract and residue values and ternary ethanol‐dichloromethane‐hexane mixtures for the basic fraction. Principal component analysis shows that the ethanol‐dichloromethane mixtures are important for extracting large quantities of the basic and organic fractions as well as of the residue and crude masses.     

Surfactant-Mediated Ultrasonic-Assisted Extraction and Purification of Antioxidants from Chaenomeles speciosa (Sweet) Nakai for Chemical- and Cell-Based Antioxidant Capacity Evaluation

In this study, a surfactant-mediated ultrasonic-assisted process was used for the first time to produce an antioxidant-enriched extract from Chaenomeles speciosa (Sweet) Nakai (C. speciosa, a popular fruit grown widely in the temperate regions of China). Ultrasonic treatment at 51 °C and 200 W for 30 min with sodium dodecyl sulfate as the surfactant led to a phenolic yield of 32.42 mg/g from dried C. speciosa powder, based on single-factor experiments, the Plackett–Burman design and the Box–Behnken design. The phenolic content increased from 6.5% (the crude extract) to 57% (the purified extract) after the purification, using LSA-900C macroporous resin. Both the crude and purified extracts exhibited a significant total reducing power and DPPH/ABTS scavenging abilities, with the purified extract being more potent. The purified extract exerted significant antioxidant actions in the tert-butyl hydroperoxide-stimulated HepG2 cells, e.g., increasing the activities of superoxide dismutase and catalase, while decreasing the reactive oxygen species and malondialdehyde levels, through the regulation of the genes and proteins of the Nrf2/Keap1 signaling pathway. Therefore, the extract from C. speciosa is a desirable antioxidant agent for the oxidative damage of the body to meet the rising demand for natural therapeutics.     

Seed Oils of Fifteen Ebenus Taxa Growing in Turkey

Oil yields and compositions from seeds of 15Ebenustaxa growing in Turkey were investigated. The yields were found between 4.0% and 13.0%. The seeds ofE. Barbigerashowed the highest whileE. Plumosavar.plumosashowed the lowest yield of oil among the 15 taxa investigated. GC/MS showed that linoleic acid (42.8-55.6%), palmitic acid (13.8-23.6%), and oleic acid (15.9-23.6%) are the main fatty acid components of all the species     

Phytochemistry and Antioxidant Activities of the Rhizome and Radix of Millettia speciosa Based on UHPLC-Q-Exactive Orbitrap-MS

The root of Millettia speciosa Champ. (MSCP) is used in folk medicine and is popular as a soup ingredient. The root is composed of the rhizome and radix, but only the radix has been used as a food. Thus, it is very important to compare the chemical components and antioxidant activities between the rhizome and radix. The extracts were analyzed by UHPLC-Q-Exactive Orbitrap-MS and multivariate analysis, and the antioxidant activities were evaluated by 2,20-azinobis (3-ethylbenzothiazo-line-6-sulfonic acid) diammonium salt (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. Ninety-one compounds were detected simultaneously and temporarily identified. Ten compounds were identified as chemical markers to distinguish the rhizome from the radix. The antioxidant activities of the radix were higher than the rhizome. Correlation analysis showed that uvaol-3-caffeate, 3-O-caffeoyloleanolic acid, and khrinone E were the main active markers for antioxidant activity, which allowed for the rapid differentiation of rhizomes and the radix. Therefore, it could be helpful for future exploration of its material base and bioactive mechanism. In addition, it would be considered to be used as a new method for the quality control of M. speciosa.     

LC/ESI/TOF-MS Characterization,Anxiolytic and Antidepressant-like Effects of Mitragyna speciosa Korth Extract in Diabetic Rats

In this study, the attenuative effects of the hydro-alcoholic extract from Mitragyna speciosa (MSE) against diabetes-induced anxiety and depression-like behaviors were examined. In addition, UPLC/ESI/TOF-MS analysis was performed to identify the phytochemical nature of MSE. DM was induced using a combination of high fructose/streptozotocin, and the diabetic rats were treated with MSE (50 and 200 mg/kg) for 5 weeks. After treatment, the animals were subjected to a forced swim test, open field test and elevated plus-maze tests. Additionally, proinflammatory cytokines and oxidative stress parameters were evaluated in the brain tissues of the rats. UPLC/ESI/TOF-MS analysis revealed that MSE is abundantly rich in polyphenolic constituents, notably flavonoid and phenolic glycosides. Behavioral tests and biochemical analyses indicated that diabetic rats showed significantly increased anxiety and depressive-like behavioral deficits, brain oxidative stress and pro-inflammatory cytokines levels (IL-1β, IL-6 and TNF-α). Treatment with MSE (50 and 200 mg/kg) significantly attenuated increased blood glucose level, depressive and anxiety-like behaviors in diabetic rats. Additionally, the antioxidant enzymes activities were markedly increased in MSE-treated animals, while TNF-α, IL-1β and IL-6 cytokines were notably suppressed. Taken together, these results suggested that MSE has potentials as antidepressant and anxiolytic-like effects and improves the brain oxido-inflammatory status in diabetic rats.     

Cytotoxicity of Callerya speciosa Fractions against Myeloma and Lymphoma Cell Lines

Callerya speciosa is widely distributed in tropical and subtropical countries and is traditionally used for preventing numerous disorders. In this study, a bioguided fractionation of ethyl acetate extract (SE) from C. speciosa root was carried out to target antioxidant and cytotoxic activities. Of the four fractions (SE1-SE4) obtained by column chromatography, SE4 had the strongest anti-radical ability in the DPPH and ABTS assays (IC₅₀ = 0.05 and 0.17 mg/mL, respectively), with results close to butylated hydroxytoluene (BHT), a common antioxidant agent. The cytotoxic activities against the selected cells were analyzed in this study by MTT assay. Accordingly, SE2, SE3, and SE4 significantly inhibited the viability of multiple myeloma cell lines, comprising U266 (IC₅₀ = 0.38, 0.09, and 0.11 mg/mL, respectively) and KMS11 (IC₅₀ = 0.09, 0.17, and 0.15 mg/mL, respectively), mantle cell lymphoma Mino (IC₅₀ = 0.08, 0.16, and 0.15 mg/mL, respectively), and the noncancerous cell line LCL (IC₅₀ = 0.40, 0.32, and 0.21 mg/mL, respectively). At a concentration of 125 µg/mL, SE2, SE3, and SE4 induced the cell apoptosis of U266 (32.2%, 53.2%, and 55.6%, respectively), KMS11 (36.9%, 40.8%, and 47.9%, respectively), Mino (36.6%, 39.8%, and 22.0%, respectively), and LCL (12.4%, 17.5%, and 23.5%, respectively) via annexin V assay. The dominant compounds detected in fractions by high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (HPLC-ESI-MS/MS), were identified as isoflavones. This is the first report describing C. speciosa as a promising natural source of antileukemia and antimyeloma agents, which may be useful for the development of blood cancer treatments.     

中药牛大力微量元素含量的测定

用电感耦合等离子体发射光谱仪(ICP)测定了湛江地区产的中药牛大力微量元素的含量,发现其Ca、Mg、Fe、Zn等元素的含量都比较丰富,相对标准偏差为0.32%~7.05%,回收率为93.0%~101%,结果令人满意。     

A high-performance liquid chromatographic method for determination of mitragynine in serum and its application to a pharmacokinetic study in rats

A simple HPLC technique for determining mitragynine levels in serum was developed. The separation system consisted of a C18 column heated to 35 degrees C, a methanol-water (80:20, v/v) mobile phase, a flow rate of 0.8 mL/min and detection in the ultraviolet at 225 nm. Mitragynine, with a retention time of 10.09 min, was well resolved from any interferences in human serum and the internal standard peak. The calibration curve was linear from 0.1 to 10 microg/mL (r = 0.9995). Extraction of mitragy-nine from alkalinized serum using diethyl ether gave a high recovery (>or=85%). The intra- and inter-day precisions of the method were 4.29-5.88%RSD and 7.06-8.45%RSD, respectively. The accuracy ranged from -9.54 to +0.67%DEV. The limit of detection was 0.03 microg/mL and the lower limit of quantification was 0.1 microg/mL. Mitragynine in the stock solution was stable during 30 days of storage at 4 degrees C. This method was successfully applied to determine the pharmacokinetic characteristics of mitragynine levels in the serum of rats after it was administered orally.     

Comparison of three chromatographic techniques for the detection of mitragynine and other indole and oxindole alkaloids in Mitragyna speciosa (kratom) plants

Leaves of the Southeast Asian plant Mitragyna speciosa are used to suppress pain and mitigate opioid withdrawal syndromes. The potential threat of abuse and ready availability of this uncontrolled psychoactive plant have led to the need for improved analytical techniques for the detection of the major active components, mitragynine and 7‐hydroxymitragynine. Three independent chromatographic methods coupled to two detection systems, GC with MS, supercritical fluid chromatography with diode array detection, and HPLC with MS and diode array detection, were compared for the analysis of mitragynine and other indole and oxindole alkaloids in M. speciosa plants. The indole alkaloids included two sets of diastereoisomers: (i) paynantheine and 3‐isopaynantheine and (ii) mitragynine, speciogynine, and speciociliatine. Two oxindole alkaloid diastereoisomers, corynoxine and corynoxine B, were also studied. The HPLC and supercritical fluid chromatography methods successfully resolved the major components with slightly different elution orders. The GC method was less satisfactory because it was unable to resolve mitragynine and speciociliatine. This separation was difficult by GC with a liquid stationary phase because these diastereoisomers differ only in the orientation of an interior hydrogen atom. The observed lack of resolution of the indole alkaloid diastereoisomers coupled with the likeness of the mass and tandem mass spectra, calls into question proposed GC methods for the analysis of mitragynine based on solely GC with MS separation and identification.     

Pigment of Ceiba speciosa (A. St.-Hil.) Flowers: Separation,Extraction, Purification and Antioxidant Activity

In this work, the extraction procedure of a natural pigment from the flower of Ceiba speciosa (A. St.-Hil.) was optimized by response surface methodology. It is the first time that the extraction of the flower pigment of C. speciosa (FPCS) has been reported, along with an evaluation of its stability and biological activity under various conditions, and an exploration of its potential use as a food additive and in medicine. Specifically, the effects of ethanol concentration, solid–liquid ratio, temperature and time on the extraction rate of FPCS were determined using a Box–Behnken design. The optimum extraction conditions for FPCS were 75% ethanol with a solid–liquid ratio of 1:75 mg/mL) at 66 °C for 39 min. The purification of FPCS using different macroporous resins showed that D101 performed best when the initial mass concentration of the injection solution was 1.50 mg/mL, resulting in a three-fold increase in color value. The yield of dry flowers was 9.75% of fresh petals and the FPCS extraction efficiency was 43.2%. The effects of light, solubility, pH, temperature, sweeteners, edible acids, redox agents, preservatives and metal ions on FPCS were also investigated. Furthermore, the characteristics of FPCS were determined by spectrophotometry at a specific wavelength using the Lambert–Beer law to correlate the mass of FPCS with its absorbance value. An acute toxicological test performed according to Horne’s method showed that FPCS is a non-toxic extract and thus may be used as a food additive or in other ingestible forms. Finally, western blotting showed that FPCS prevents lipopolysaccharide-induced hippocampal oxidative stress in mice. The study suggests that FPCS may function as an antioxidant with applications in the food, cosmetics and polymer industries.