Extraction, recovery, and characterization of lignin from industrial corn stover lignin cake.

Lignin utilization in value-added co-products is an important component of enabling cellulosic biorefinery economics. However, aqueous dilute acid pretreatments yield lignins with limited applications due to significant modification during pretreatment, low solubility in many solvents, and high content of impurities (ash, insoluble polysaccharides). This work addresses these challenges and investigates the extraction and recovery of lignins from lignin-rich insoluble residue following dilute acid pretreatment and enzymatic hydrolysis of corn stover using three extraction approaches: ethanol organosolv, NaOH, and an ionic liquid. The recovered lignins exhibited recovery yields ranging from 30% for the ionic liquid, 44% for the most severe acid ethanol organosolv condition tested, and up to 86% for the most severe NaOH extraction condition. Finally, the fractional solubilities of different recovered lignins were assessed in a range of solvents and these solubilities were used to estimate distributions of Hildebrand and Hansen solubility parameters using a novel approach.

Gastroprotective potential of hydro-alcoholic extract of Pattanga (Caesalpinia sappan Linn.).

ETHNO-PHARMACOLOGICAL RELEVANCE: Pattanga is botanically equated as Caesalpinia sappan Linn. (Family: Caesalpiniaceae) and is used in Ayurveda system of medicine since ages. According to Ayurveda, useful part is Heartwood, which is bitter, astringent and acrid and is useful in vitiated conditions of vata and pitta, burning sensation, wounds, ulcers, leprosy, skin diseases, menorrhagia, leucorrhea, and diabetes. It is used as a major ingredient in Ayurvedic formulations and preparations like Patrangasava, Chandanadya Thalia, and Karpuradyarka. AIM OF THE STUDY: The present study is planned to evaluate the gastroprotective activity of the selected Ayurvedic drug using three different in vivo gastric ulcer models, so as to provide scientific evidence for the Ayurvedic claims. MATERIALS AND METHODS: For this study, Wistar albino rats fasted overnight were selected. The hydroalcoholic extract of Caesalpinia sappan heartwood at the dose level 250 and 500mg/kg body weight was selected and administered orally before necrotizing agents. Antioxidant and antiulcer parameters were evaluated and the stomach samples were subjected for histopathological studies. In addition, PGE2 estimation and protein expressions of COX-1, COX-2 and iNOS were analyzed by Western blot. The plant extract was subjected to LCMS/MS analysis. In addition, Cytoprotective effect in isolated gastric mucosal cells, TUNEL Assay, Acid neutralizing capacity assay, H+/K+ ATPase inhibitory assay were performed. RESULTS: The ulcer protection was found to be 92%, 86% and 64% against ethanol, NSAID and pylorus ligation induced ulcer respectively. The hydro-alcoholic extract of C. sappan heartwood exhibited cytoprotective effect with 76.82% reduction against indomethacin-induced cytotoxicity at the concentration of 25µg/ml. C. sappan showed 63.91% inhibition in H+/K+ ATPase inhibitory assay at the concentration 500µg/ml. CONCLUSIONS: Our results depict that Caesalpinia sappan heartwood possesses gastroprotective activity, possibly mediated through cytoprotection and antioxidant mechanisms. The data obtained in the present study provides scientific support for the traditional use of Caesalpinia sappan in the management of peptic ulcer.

Do stress-whitening and optical clearing of collagenous tissue occur by the same mechanism?

Bone is biphasic with an organic matrix and an inorganic mineral component. As we age bone's susceptibility to fracture increases. It has been shown that there is no change in mean mineralization with aging, but bone nevertheless becomes less tough. This aging effect is therefore likely related to the organic phase. Under mechanical loading, immediately prior to failure, bone has been observed to visually become more opaque and has been termed stress-whitening. Stress-whitening is known to make materials tougher. The goal of this investigation was to investigate stress-whitening in the collagenous matrix of bone. Hydrogen bonds play a key role in collagen stability and we hypothesize that changes in hydrogen bonding will significantly affect matrix stiffness, toughness and stress whitening. Demineralized bone specimens were loaded in tension and stress-whitening was monitored. The effect of hydrogen bonding on mechanical properties and stress-whitening process was probed by altering the Hansen's hydrogen bonding parameter (δh) of the immersing solution. The Hansen's hydrogen bonding parameter of the immersing fluid affected the morphology, mechanical properties and stress whitening of specimens. Specimens were visually whiter in the absence of mechanical load in low δh solvents (the specimens solvent-whitened). Both the observed stress-whitening and solvent-whitening were reversible and repeatable processes. The observed solvent-whitening that occurred without the presence of load was consistent with solvent-induced optical clearing (the opposite of whitening) in skin caused by collagen fibril swelling. Stress whitening and solvent whitening can be explained by a common mechanism, collagen fibril densification and thinning, leading to an increased distinction between the collagen fibrillar phase and immersing fluid, ultimately leading to more scattering. Bones may be at a greater risk for fracture as we age because solubility of the matrix changes, thus making the collagen less hydrated (and more brittle) even in the same solvent.

Antinociceptive, anti-inflammatory and antiulcerogenic activities of ethanol root extract of Strophanthus hispidus DC (Apocynaceae).

BACKGROUND: Strophanthus hispidus DC (Apocynaceae) is a medicinal plant widely used in traditional African medicine in the treatment of rheumatic afflictions, ulcer, conjunctivitis, leprosy and skin diseases. This study sought to investigate the antinociceptive, anti-inflammatory and antiulcer properties of the ethanol root extract of S. hispidus. METHODS: Antinociceptive activity was evaluated using acetic acid-induced writhing and formalin tests in mice. The carrageenan- and egg albumin-induced rat paw edema tests were used to investigate the anti-inflammatory actions, whereas the antiulcer activity was investigated using ethanol-, HCl- and pyloric ligation-induced gastric ulcer models in rats. RESULTS: S. hispidus [100-800 mg/kg orally (po)] produced significant (p<0.05) inhibition of writhing reflex with peak effect of 74.13% inhibition observed at 800 mg/kg. Similarly, S. hispidus significantly (p<0.05) attenuated formalin-induced early and late phase of nociception with peak effect of 61.84% and 89.43%, respectively, at 200 mg/kg. S. hispidus (25-800 mg/kg po) caused significant (p<0.05) inhibition of edema development in the carrageenan and egg albumin models with peak effect (93.40% and 90.10% inhibition of edema formation) observed at 50 mg/kg. With respect to antiulcer activity, S. hispidus (100-800 mg/kg) showed potent antiulcer activity with respective peak effects of 96% (ethanol-induced), 99% (HCl-induced) and 70.60% inhibition of ulcer. CONCLUSIONS: The findings in this study suggest that the ethanol root extract of S. hispidus possesses antinociceptive, anti-inflammatory and antiulcerogenic activities. This justifies the use of the extract in folklore medicine for the treatment of ulcer and inflammatory disorders.

Nephroprotective activity of Cocculus hirsutus leaf extract in 5/6 nephrectomized rat model.

BACKGROUND: Cocculus hirsutus, commonly known as broom creeper, belongs to the family Menispermaceae. It is widely used in folk medicine to treat leprosy, skin diseases, dyspepsia, etc. Hence, an effort has been made to investigate the nephroprotective potential of C. hirsutus. METHODS: The nephroprotective activity of ethanolic C. hirsutus leaf extract (ECHE) in the 5/6 nephrectomized rat model was investigated. Different parameters like postoperative survival rate, change in body weight and levels of red blood corpuscles (RBCs), hemoglobin (Hb), triglyceride, cholesterol, creatinine, urea and uric acid were estimated in experimental rats. RESULTS: The findings revealed the postoperative survival rate of rats in the investigated novel method to be 100%. Change in the body weight of ECHE-treated groups I and II was found to be 32 and 30 g, respectively. These values suggested that ECHE treatment normalized the elevated body weight levels in experimental rats. Furthermore, ECHE treatment normalized the decreased RBC levels and the elevated Hb, triglyceride, cholesterol, creatinine, urea and uric acid levels in experimental rats. CONCLUSIONS: The results indicate that C. hirsutus has strong nephroprotective activity. However, further scrutiny is essential for isolation and characterization of the active components that can be employed to allay various human maladies.

Identification of active compounds from Caesalpinia sappan L. extracts suppressing IL-6 production in RAW 264.7 cells by PLS.

ETHNOPHARMACOLOGICAL RELEVANCE: Caesalpinia sappan L. is distributed in Southeast Asia and also used as herbal medicine for the treatment of various diseases such as burning sensations, leprosy, dysentery, osteoarthritis and rheumatoid arthritis (RA). The overproduction of IL-6 plays an important role in the prognosis of RA, but the active compounds from the extracts of Caesalpinia sappan L. suppressing IL-6 production remain unknown. AIMS OF THE STUDY: Identifying the main active compounds of Caesalpinia sappan L. extracts inhibiting the IL-6 production in LPS-stimulated RAW 264.7 cells by partial least squares (PLS). MATERIALS AND METHODS: Sixty-four samples with different proportions of compounds were prepared from Caesalpinia sappan L. by supercritical CO2 fluid extraction (SCFE) and refluxing. Each of 64 samples was applied to RAW 264.7 cells with LPS to evaluate whether IL-6 production by LPS is affected by addition of each sample. The IL-6 production in medium was determined by ELISA and the inhibitory activity of each sample was analyzed. In addition, the fingerprints of these 64 samples were also established by ultra-performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-MS). We used the PLS, a simplified method, to evaluate the results from IL-6 production and fingerprints. RESULTS: Each of 64 samples markedly suppressed LPS-induced IL-6 production in RAW cells. The fingerprints by UPLC-MS clearly revealed variations among 64 samples produced in different extract conditions. The PLS analysis with IL-6 production and fingerprints by UPLC-MS suggested that the peaks 71, 93, 150, 157, 168 have more influence on the inhibitory activity of Caesalpinia sappan L. extracts. The peaks 71, 93, 150 are likely representing sappanone A, protosappanin E and neoprotosappanin, respectively. The peaks 157 and 168 are still at large. CONCLUSION: This is the first report that sappanone A, protosappanin E, neoprotosappanin and two unidentified compounds can be considered as possible active compounds that might inhibit IL-6 production. Further studies are needed to confirm the effectiveness of these five compounds on IL-6 production and possible mechanism.

Combretum leprosum Mart. (Combretaceae): potential as an antiproliferative and anti-inflammatory agent.

ETHNOPHARMACOLOGICAL RELEVANCE: Combretum leprosum is a species that is popularly used in Brazil as a healing agent to treat skin problems and lesions. In this study we investigated the possible potential of this extract to treat inflammatory and hyperproliferative skin conditions. MATERIALS AND METHODS: Classical models of skin inflammation such as TPA- and croton oil-induced mouse ear oedema were applied in order to verify the potential topical anti-inflammatory activity of the ethanolic extract from flowers of Combretum leprosum. RESULTS: Topical application of ethanolic extract promoted a dose-dependent inhibition of phorbol ester-induced ear oedema, reduced myeloperoxidase activity and IL-6 tissue levels with inhibition comparable to dexamethasone (positive control). Histological and immunohistochemical analysis revealed that ethanolic extract also suppressed cell infiltration. Ethanolic extract altered inflammatory parameters on a chronic skin inflammation model induced by repeated applications of croton oil, decreasing ear oedema, epidermal hyperproliferation and cell infiltration. In addition, immunohistochemical analysis showed that the extract decreased PCNA expression on the epidermis. CONCLUSION: Taken together, these results suggest that the extract from flowers of Combretum leprosum could be considered as a new potential tool for the treatment of several skin inflammatory diseases since it reversed the skin inflammatory and hyperproliferative process in a very significant manner. Further investigations are needed in order to verify the cellular mechanism and safety of Combretum leprosum extract.

Effect of ethanol on the sorption of four targeted wine volatile compounds in a polyethylene film.

The objectives of this study were to demonstrate that the presence of ethanol in a solution containing two esters and two aromatic alcohols has several consequences on the sorption of these compounds into polyethylene (PE) film. First, sorption of ethanol into the PE film occurred at the same time as water and reached 8 kg m(-3) using 12% v/v of ethanol. This sorption was associated with an increase in PE crystallinity, which may have prevented the sorption of volatile compounds despite their strong affinity with PE film, as evaluated by Hansen solubility parameters. Moreover, increasing the ethanol concentration increased the solubility of the four volatile compounds. In the case of aromatic alcohols, the sorption was decreased in the presence of ethanol as expected. In the case of esters, as their hydrolysis was substantial in the presence of water, the consequence was a higher sorption into the PE film in the presence of ethanol than in its absence. Nevertheless, the sorption also depended on the concentration of ethanol and the heterogeneity of the ethanol-water mixture as well as the presence of other volatile compounds, as in the case of 4-ethylphenol. In conditions simulating wine packaging, losses of volatile compound by sorption and by permeation estimated after only 5 days of contact varied between 0.08 and 25% for 2-phenylethanol and ethyl hexanoate, respectively.

Evaluation of antimicrobial activity of ethanol extract and compounds isolated from Trichodesma indicum (Linn.) R. Br. root.

ETHNOPHARMACOLOGICAL RELEVANCE: The roots are reportedly used to treat diarrhoea, dysentery, leprosy, skin diseases and fever. AIM OF STUDY: The aim of present study was to investigate the in vitro antimicrobial potential of ethanol extract of Trichdesma indicum root, and its purified compounds and to validate scientifically its use in traditional medicine. MATERIAL AND METHODS: The root of Trichdesma indicum was extracted with ethanol and subjected to chromatographic separation for isolation of phytochemical compounds. Structures of isolated compounds were elucidated by spectroscopic methods. The antimicrobial activities of the ethanol extract of T. indicum and isolated compounds were primarily evaluated by a disc diffusion test. The anti-microbial efficacy of the ethanol extract or isolated compounds was then assessed in vitro by determining minimal inhibition concentration (MIC) and minimal bactericidal or fungicidal concentration (MBC/MFC). RESULTS: n-Decanyl laurate (1), n-tetradecanyl laurate (2), n-nonacosanyl palmitate (3), stigmast-5-en-3ß-ol-21(24)-olide (4), n-pentacos-9-one (5), n-dotriacont-9-one-13-ene (6), stigmast-5-en-3ß-ol-23-one (7) and lanast-5-en-3ß-D-glucopyranosyl-21 (24)-olide (8) were isolated from ethanol extract of T.indicum. The ethanol extract and isolated compounds (1-8) showed varying degrees of antimicrobial activities. The ethanol extract exhibited potent growth inhibitory activity against S. aureus, B. subtilis and C. albicans with an MIC value of 19.2 µg/ml. Among all the isolated compounds, lanast-5-en-3ß-D-glucopyranosyl-21 (24)-olide (8) displayed strongest antibacterial activity against S. aureus with MIC value of 2.4 µg/ml. CONCLUSIONS: The results of present study provide ground basis for the potential use of the ethanol extract Trichodesma indicum root as well as the some of the isolated compounds in the treatment of infections associated with the studied microorganisms.

Effect of nutrient supplementation of crude or detoxified concentrated distilled grape marc hemicellulosic hydrolysates on the xylitol production by Debaryomyces hansenii.

Biosynthesis of xylitol using the yeast Debaryomyces hansenii NRRL Y-7426 was carried out using distilled grape marc (DGM) hemicellulosic hydrolysates directly concentrated by vacuum evaporation or after detoxification with activated charcoal. The effect of nutrient supplementation with vinasses, corn steep liquor (CSL) or commercial nutrients was explored. Using crude concentrated hemicellulosic hydrolysates, the maximum xylitol concentration, 11.3 g/L, was achieved after 172 hr (Q ( xylitol ) = 0.066 g/L-hr; Y ( xylitol ) (/SC) = 0.21 g/g); meanwhile, using detoxified concentrated hydrolysates, the concentration increased up to 19.7 g/L after 72 hr (Q ( xylitol ) = 0.274 g/L-hr; Y ( xylitol ) (/SC) = 0.38 g/g). On the other hand, using crude or detoxified hydrolysates, the xylose-to-xylitol bioconversion was strongly affected by the addition of nutrients, suggesting that these hydrolysates present essential nutrients favouring the growth of D. hansenii.

Optimization of subcritical fluid extraction of bioactive compounds using Hansen solubility parameters.

Process engineering operations in food and nutraceutical industries pertaining to the design of extraction of value-added products from biomass using pressurized liquids involve a careful selection of the solvent and optimal temperature conditions to achieve maximum yield. Complex molecular structure and limited physical property data in the literature of biological solutes extracted from biomass compounds have necessitated the process modeling of such operations. In this study, we have applied the Hansen 3-dimensional solubility parameter concept to optimize the extraction of molecularly complex solutes using subcritical fluid solvents. Hansen solubility spheres characterized by the relative energy differences (RED) have been used to characterize and quantify the solute-subcritical solvent interactions as a function of temperature. The solvent power of subcritical water and compressed hydroethanolic mixtures above their boiling points has been characterized using the above-mentioned method. The use of group contribution methods in collaboration with computerized algorithms to plot the Hansen spheres provides a quantitative prediction tool for optimizing the design of extraction conditions. The method can be used to estimate conditions for solute-solvent miscibility, an optimum temperature range for conducting extractions under pressurized conditions, and approximate extraction conditions of solutes from natural matrices.