Dimethyl Sulfoxide Extraction Research Articles

Enhancement of terephthalic acid recovered from PET waste using a combination of citric acid and dimethyl sulfoxide extraction

This study aimed to develop an eco-friendly, cost-efficient, and practically viable method for extracting terephthalic acid (H2BDC) from polyethylene terephthalic (PET) waste. Dimethyl sulfoxide (DMSO) was combined with either citric acid (C6H8O7) or H2SO4 to enhance the particle size of H2BDC, and the optimum conditions during the acidification step were determined. Additionally, response surface methodology was employed to examine the influence and interaction of extractant (NaOH) concentration, hydrolysis temperature, and time on the optimal H2BDC yield and recovery ratio. Experimental results demonstrated that NaOH concentration significantly impacted both H2BDC yield and recovery ratio, surpassing the effects of hydrolysis temperature and time. Under optimal conditions involving a temperature of 200 °C and a 12 h reaction time with 5% NaOH, the model predicted a 100% yield and recovery ratio, which closely matched the experimental results of 99% and 100% for yield and recovery ratio, respectively. To enhance particle size, a combination of DMSO and C6H8O7 was more effective than H2SO4. The maximum particle size achieved was 57.4 µm under the following optimum conditions: premixing 5 M C6H8O7 with DMSO at a 35:75 mL ratio and maintaining a reaction temperature of 75 °C for 40 min. The study demonstrated the stability and consistency of the method. The H2BDC yield remained between 96 and 98% with high purity over eight consecutive cycles of using the DMSO and C6H8O7 mixture. The findings highlight the importance of integrating C6H8O7 and DMSO to enhance H2BDC quality, meeting commercial product criteria with evidence of high purity and large particle size. This method presents a promising solution for extracting H2BDC from PET waste, with potential implications for the recycling industry and a positive environmental impact.

The effects of holocellulose treatments on the yield and structure of dimethylsulfoxide-extracted birchwood xylans

Xylans with naturally occurring acetylation are vital to the plant cell wall functionalities and are also demonstrating excellent dispersing properties which can be applied in fine chemicals and advanced materials. The acetyl structure of xylan can be maintained by dimethyl sulfoxide (DMSO) extraction. A disadvantage of DMSO extraction is that its yield is much lower than that of alkali extraction, which is unfavorable to the research and application of xylans. In this study, sodium chlorite was used as the delignification method to produce holocellulose from birch wood. The effects of subsequent holocellulose solvent exchange, drying and ball-milling on xylan DMSO extraction were studied. The differences of three successive DMSO extractions from the same holocellulose sample and gradient xylan precipitation from DMSO were also evaluated. The majority of xylans are extracted in the first two extractions, while the xylans extracted in the third extraction usually had low yields and low molecular weights. Xylans extracted directly from aqueously wet holocellulose without drying had a very low yield of less than 1%, while xylans extracted from freeze or vacuum dried holocellulose have total yields of 4–9%. T-butanol or ether washed holocellulose shows slightly lower xylan yield than water washed holocellulose. Ball milled holocellulose has twice the xylan yield of holocellulose sample without ball milling. For gradient xylan precipitation, more than 95% of the xylans precipitate in 50% and 70% ethanol. Gradient ethanol precipitation fractionates xylans by molecular weight, but does not fractionate xylans by degree of acetylation. This study is significant for promoting the researches and applications of DMSO extraction of xylan.

High-yield dimethyl ether-based recovery of astaxanthin and fatty acids directly from wet Haematococcus pluvialis

Efficient and eco-friendly methods for extracting bioactive molecules are crucial for achieving economically viable microalgae biorefinery. In this study, high-value-added intracellular bioactive compounds were efficiently recovered directly from the red cysts of wet Haematococcus pluvialis. An integrated processes involving liquid dimethyl ether (l-DME) extraction at 45 °C and 2 MPa for 90 min and subsequent dimethyl sulfoxide (DMSO) extraction at 65 °C for 5 min enabled almost complete recovery of astaxanthin (99.6%) and total fatty acids (99.8%) from wet H. pluvialis cyst cells. Notably, this green and sustainable strategy did not involve high-cost and energy-intensive drying, or cell wall-disruption steps. l-DME extraction produced a solvent-free, dry extract with high astaxanthin (43.9 mg g−1 dry extract) and essential fatty acids (ω3, ω6, and ω9; 290.1 mg g−1 dry extract) contents; whereas the astaxanthin and fatty acid contents of the DMSO extract produced from raw H. pluvialis cyst cells (freeze-dried and ball-milled at 200 rpm for 30 min) were 1.5- and 1.7-fold lower, respectively. The dry extract also exhibited 1.6-fold greater antioxidant activity than the DMSO extract, thereby indicating its potential for direct application in nutraceutical, pharmaceutical, and cosmeceutical formulations with high bioactivity and safety.

Structural characterization of corn fiber hemicelluloses extracted by organic solvent and screening of degradation enzymes

An integrated treatment coupling peracetic acid delignification, dimethyl sulfoxide extraction, and ethanol precipitation were performed to isolate hemicellulose from de-starched corn fiber. Based on chemical composition, molecular weight distribution, methylation, and nuclear magnetic resonance spectroscopy, it is proposed that hemicelluloses in corn fiber were composed of two polysaccharides, glucuronoarabinoxylan (about 80 %) and xyloglucan (about 20 %). Xylose (about 46 %) and arabinose (about 32 %) were the main components in glucuronoarabinoxylan. More than half of the xylose units in the glucuronoarabinoxylan backbone chain were substituted at O-2 and/or O-3 by various monomers or oligomeric side chains. Based on structure analysis, five hemicellulases were selected and added to Penicillium oxalicum MCAX enzymes for enzymatic hydrolysis of corn fiber. The results showed that the addition of hemicellulases increased the sugar yield of corn fiber. These results demonstrate the effectiveness of enzyme consortium constructed by elucidating the chemical structure of hemicellulose in corn fiber for the degradation of corn fiber and also provide a general solution for the rational construction of targeted and efficient enzyme systems for the degradation of lignocellulosic biomass.

Optimizing yield and chemical compositions of dimethylsulfoxide-extracted birchwood xylan

Dimethylsulfoxide (DMSO) extraction is commonly used to study the chemical structures of original xylan in the plant cell wall, since the DMSO can preserve the original structure of the xylan as much as possible during the extracting process. In addition, the DMSO-extracted xylans have unique properties allowing their potential applications in emulsifying or filming materials. However, the yield of DMSO-extracted xylan is always low and the effects of different DMSO extraction conditions on the chemical compositions of xylan have not been fully studied, which greatly hinders its researches and applications. In this study, we have found that extensive delignification before DMSO extraction results in destruction of lignin-carbohydrate complex (LCC), leading to xylan yield and xylose unit content increased by up to 220% and 20%, respectively. Tert-butanol washing of the holocellulose can further increase the DMSO extracted xylan yield by ∼10%. The yield of xylan extracted by the DMSO at 80 °C for 7 h was obviously higher than that at room temperature for 3 d by 30%–40%. Thermal analysis showed that the xylans extracted at different conditions had thermal stability without obvious differences. The results indicate that the DMSO-extracted xylan with a high yield, a high purity and a high degree of acetylation can be extracted at a high delignification level, a high reaction temperature and a short reaction time. This study is of great significance for studying xylan structure-property relationships and promoting the applications of DMSO-extracted xylan.

Extraction of acetylated glucuronoxylans and glucomannans from Okoume (Aucoumea klaineana Pierre) sapwood and heartwood by steam explosion

Aucoumea klaineana Pierre (okoume) is a tropical hardwood of industrial importance, used for veneers and plywood production. Various conditions of Steam Explosion (SE) treatment (temperature, duration, pH) have been applied on okoume co-products, sapwood (SW) and heatwood (HW) for the extraction of polymeric hemicelluloses. Conventional extractions with alkali or dimethyl sulfoxide (DMSO) were also performed from delignified SW and HW for comparison. The hemicelluloses extracts have been characterized by HSQC-NMR, FTIR, size exclusion chromatography. The acetyl contents have been determined by chromatography after saponification and from HSQC spectra. Using basic impregnation prior to SE (NaOH-SE), high molecular mass glucuronoxylans (GX, Mw = 65–100 KDa) were recovered in 3–6 % % yields. A neutral impregnation (H2O-SE) produced O2 and/or O3 acetylated GX and glucomannans (GM), preserving the acetyl moieties in a greater extent compared to the mild DMSO extraction (% Ac ≈ 6.6–8.4 % and 1.0–4.1 % w/w respectively). This study provides insights on the composition of SW and HW okoume with a view to further innovative processing and valorization including the preparation of films with advanced properties for food packaging.

In Vitro Bioavailability of the Hydrocarbon Fractions of Dimethyl Sulfoxide Extracts of Petroleum Substances.

Determining the in vitro bioavailable concentration is a critical, yet unmet need to refine in vitro-to-in vivo extrapolation for unknown or variable composition, complex reaction product or biological material (UVCB) substances. UVCBs such as petroleum substances are commonly subjected to dimethyl sulfoxide (DMSO) extraction in order to retrieve the bioactive polycyclic aromatic compound (PAC) portion for in vitro testing. In addition to DMSO extraction, protein binding in cell culture media and dilution can all influence in vitro bioavailable concentrations of aliphatic and aromatic compounds in petroleum substances. However, these in vitro factors have not been fully characterized. In this study, we aimed to fill in these data gaps by characterizing the effects of these processes using both a defined mixture of analytical standards containing aliphatic and aromatic hydrocarbons, as well as 4 refined petroleum products as prototypical examples of UVCBs. Each substance was extracted with DMSO, and the protein binding in cell culture media was measured by using solid-phase microextraction. Semiquantitative analysis for aliphatic and aromatic compounds was achieved via gas chromatography-mass spectrometry. Our results showed that DMSO selectively extracted PACs from test substances, and that chemical profiles of PACs across molecular classes remained consistent after extraction. With respect to protein binding, chemical profiles were retained at a lower dilution (higher concentration), but a greater dilution factor (ie, lower concentration) resulted in higher protein binding in cell medium, which in turn altered the ultimate chemical profile of bioavailable PACs. Overall, this case study demonstrates that extraction procedures, protein binding in cell culture media, and dilution factors prior to in vitro testing can all contribute to determining the final bioavailable concentrations of bioactive constituents of UVCBs in vitro. Thus, in vitro-to-in vivo extrapolation for UVCBs may require greater attention to the concentration-dependent and compound-specific differences in recovery and bioavailability.

Development of SPAD value-based linear models for non-destructive estimation of photosynthetic pigments in wheat (Triticum aestivum L.)

Chlorophyll content in leaf which can be measured and expressed as SPAD values, is a good indicator of plant health. The present study aimed to develop linear models for studying relationship between SPAD values and spectrophotometrically determined chlorophylls and carotenoid contents in leaves of 468 wheat genotypes. The SPAD values were taken in the flag leaves at heading stage. Chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll (total Chl) and total carotenoids (total Car) contents were assessed using dimethyl sulfoxide (DMSO) extraction method. To estimate photosynthetic pigment content in leaves, SPAD value-based linear models were developed and Chla (Chlb = 0.0690 x SPAD Value - 1.082), Chla (Chlb = 0.021 x SPAD Value 0.396), total Chl (Total chlorophyll = 0.090 x SPAD Value - 1.477) and total carotenoids (Total Carotenoids = 0.013 x SPAD Value 0.074) were estimated. The present study reports development of linear model for quantification of the photosynthetic pigment content in wheat leaves using chlorophyll meter.

Accurate quantification of astaxanthin from Haematococcus pluvialis using DMSO extraction and lipase-catalyzed hydrolysis pretreatment

Haematococcus pluvialis accumulates high contents of astaxanthin and is considered as an important source of natural astaxanthin production. Fast and accurate quantification of astaxanthin is needed to evaluate the quality of microalgal sources for astaxanthin extraction and to monitor the H. pluvialis cultivation process. In the present study, a less-invasive method for astaxanthin accurate quantification was developed. Dimethyl sulfoxide (DMSO) was used as an extraction solvent to improve the membrane permeability of H. pluvialis. Cholesterol esterase was used to hydrolyze astaxanthin esters via incubation at 36 °C for 30 min. The detection of astaxanthin after hydrolysis was achieved by high performance liquid chromatography (HPLC) within 10 min using a C18 column. The astaxanthin content measurements after enzymatic pretreatment were 1.5–1.7 times higher than that measured after saponified pretreatment. DMSO solvent extraction combined with lipase-catalyzed hydrolysis followed by HPLC detection is proposed for the accurate quantification of astaxanthin in H. pluvialis.

Short communication: Aqueous-acetone extraction improves the drawbacks of using dimethylsulfoxide as solvent for photometric pigment quantification in Quercus ilex leaves

Aim of study: We evaluated the use of dimethylsulfoxide (DMSO) for the photometric determination of chlorophyll (Chla and Chlb) and carotenoids in Quercus ilex L. leaves by comparative analysis using aqueous-acetone extraction.Area of study: a Q. ilex dehesa in Las Majadas del Tietar, Cáceres, SpainMaterial and methods: Q. ilex leaves were sampled during two vegetative periods. Field SPAD-502 Chlorophyll measurements and photometric chlorophyll determinations were performed. Two procedures were used: extraction of intact foliar discs at 65ºC with DMSO and fine foliar powder with cold aqueous-acetone.Main results: DMSO produced Chlb overestimation and different fitting performance for SPAD vs pigment calibrations (R2=0.64, RMSE=0.20, p<0.0001 for Chla (µg/cm2); R2=0.33, RMSE=0.23, p<0.0001 for Chlb (µg/cm2) and R2=0.50, RMSE=0.23, p<0.0001 for carotenoids (µg/cm2)). Aqueous-acetone provided more accurate predictions (R2=0.90, RMSE=0.16, p<0.0001 for Chla and R2=0.91, RMSE=0.16, p<0.0001 for Chlb, R2=0.90, RMSE=0.02, p<0.0001 for carotenoids) and mean ratio Chla/Chlb=3.6 inside the range for sun exposed leaves.Research highlights: Oxidizing conditions and polyphenol concentrations in Q. ilex leaves generated brown colorations in the DMSO extraction procedure, interfering with the photometric measurements in the red-orange region. Aqueous-acetone extraction was free from interference. DMSO should be avoided for pigment determination in Q. ilex leaves or when comparing different tree species.

In vitro–ex vivo correlations between a cell-laden hydrogel and mucosal tissue for screening composite delivery systems

Composite delivery systems where drugs are electrospun in different layers and vary the drug stacking-order are posited to affect bioavailability. We evaluated how the formulation characteristics of both burst- and sustained-release electrospun fibers containing three physicochemically diverse drugs: dapivirine (DPV), maraviroc (MVC) and tenofovir (TFV) affect in vitro and ex vivo release. We developed a poly(hydroxyethyl methacrylate) (pHEMA) hydrogel release platform for the rapid, inexpensive in vitro evaluation of burst- and sustained-release topical or dermal drug delivery systems with varying microarchitecture. We investigated properties of the hydrogel that could recapitulate ex vivo release into nonhuman primate vaginal tissue. Using a dimethyl sulfoxide extraction protocol and high-performance liquid chromatography analysis, we achieved >93% recovery from the hydrogels and >88% recovery from tissue explants for all three drugs. We found that DPV loading, but not stacking order (layers of fiber containing a single drug) or microarchitecture (layers with isolated drug compared to all drugs in the same layer) impacted the burst release in vitro and ex vivo. Our burst-release formulations showed a correlation for DPV accumulation between the hydrogel and tissue (R2= 0.80), but the correlation was not significant for MVC or TFV. For the sustained-release formulations, the PLGA/PCL content did not affect TFV release in vitro or ex vivo. Incorporation of cells into the hydrogel matrix improved the correlation between hydrogel and tissue explant release for TFV. We expect that this hydrogel-tissue mimic may be a promising preclinical model to evaluate topical or transdermal drug delivery systems with complex microarchitectures.

Isolation and characterization of acetylated glucuronoarabinoxylan from sugarcane bagasse and straw

Sugarcane bagasse and straw are generated in large volumes as by-products of agro-industrial production. They are an emerging valuable resource for the generation of hemicellulose-based materials and products, since they contain significant quantities of xylans (often twice as much as in hardwoods). Heteroxylans (yields of ca 20% based on xylose content in sugarcane bagasse and straw) were successfully isolated and purified using mild delignification followed by dimethyl sulfoxide (DMSO) extraction. Delignification with peracetic acid (PAA) was more efficient than traditional sodium chlorite (NaClO2) delignification for xylan extraction from both biomasses, resulting in higher extraction yields and purity. We have shown that the heteroxylans isolated from sugarcane bagasse and straw are acetylated glucuronoarabinoxylans (GAX), with distinct molecular structures. Bagasse GAX had a slightly lower glycosyl substitution molar ratio of Araf to Xylp to (0.5:10) and (4-O-Me)GlpA to Xylp (0.1:10) than GAX from straw (0.8:10 and 0.1:10 respectively), but a higher degree of acetylation (0.33 and 0.10, respectively). A higher frequency of acetyl groups substitution at position α-(1→3) (Xyl-3Ac) than at position α-(1→2) (Xyl-2Ac) was confirmed for both bagasse and straw GAX, with a minor ratio of diacetylation (Xyl-2,3Ac). The size and molecular weight distributions for the acetylated GAX extracted from the sugarcane bagasse and straw were analyzed using multiple-detection size-exclusion chromatography (SEC-DRI-MALLS). Light scattering data provided absolute molar mass values for acetylated GAX with higher average values than did standard calibration. Moreover, the data highlighted differences in the molar mass distributions between the two isolation methods for both types of sugarcane GAX, which can be correlated with the different Araf and acetyl substitution patterns. We have developed an empirical model for the molecular structure of acetylated GAX extracted from sugarcane bagasse and straw with PAA/DMSO through the integration of results obtained from glycosidic linkage analysis, 1H NMR spectroscopy and acetyl quantification. This knowledge of the structure of xylans in sugarcane bagasse and straw will provide a better understanding of the isolation-structure-properties relationship of these biopolymers and, ultimately, create new possibilities for the use of sugarcane xylan in high-value applications, such as biochemicals and bio-based materials.

Structural characterization and anti-inflammatory activity of a linear β-d-glucan isolated from Pleurotus sajor-caju

Glucans comprise an important class of polysaccharides present in basidiomycetes with potential biological activities. A (1→3)-β-d-glucan was isolated from Pleurotus sajor-caju via extraction with hot water followed by fractionation by freeze-thawing and finally by dimethyl sulfoxide extraction. The purified polysaccharide showed a 13C-NMR spectrum with six signals consisting of a linear glucan with a β-anomeric signal at 102.8ppm and a signal at 86.1ppm relative to O-3 substitution. The other signals at 76.2, 72.9, 68.3, and 60.8ppm were attributed to C5, C2, C4, and C6, respectively. This structure was confirmed by methylation analysis, and HSQC studies. The β-d-glucan from P. sajor-caju presented an immunomodulatory activity on THP-1 macrophages, inhibited the inflammatory phase of nociception induced by formalin in mice, and reduced the number of total leukocytes and myeloperoxidase levels induced by LPS. Taken together, these results demonstrate that this β-d-glucan exhibits a significant anti-inflammatory activity.

A high-throughput biliverdin assay using infrared fluorescence.

Biliverdin is an intermediate of heme degradation with an established role in veterinary clinical diagnostics of liver-related diseases. The need for chromatographic assays has so far prevented its wider use in diagnostic laboratories. The current report describes a simple, fast, high-throughput, and inexpensive assay, based on the interaction of biliverdin with infrared fluorescent protein (iRFP) that yields functional protein exhibiting infrared fluorescence. The assay is linear in the range of 0-10 µmol/l of biliverdin, has a limit of detection of 0.02 μmol/l, and has a limit of quantification of 0.03 µmol/l. The assay is accurate with relative error less than 0.15, and precise, with coefficient of variation less than 5% in the concentration range of 2-9 µmol/l of biliverdin. More than 95% of biliverdin was recovered from biological samples by simple dimethyl sulfoxide extraction. There was almost no interference by hemin, although bilirubin caused an increase in the biliverdin concentration, probably due to spontaneous oxidation of bilirubin to biliverdin. The newly developed biliverdin assay is appropriate for reliable quantification of large numbers of samples in veterinary medicine.

Hematopoietic SCT with cryopreserved grafts: adverse reactions after transplantation and cryoprotectant removal before infusion

Transplantation of hematopoietic stem cells (HSCs) has been successfully developed as a part of treatment protocols for a large number of clinical indications, and cryopreservation of both autologous and allogeneic sources of HSC grafts is increasingly being used to facilitate logistical challenges in coordinating the collection, processing, preparation, quality control testing and release of the final HSC product with delivery to the patient. Direct infusion of cryopreserved cell products into patients has been associated with the development of adverse reactions, ranging from relatively mild symptoms to much more serious, life-threatening complications, including allergic/gastrointestinal/cardiovascular/neurological complications, renal/hepatic dysfunctions, and so on. In many cases, the cryoprotective agent (CPA) used-which is typically dimethyl sulfoxide (DMSO)-is believed to be the main causal agent of these adverse reactions and thus many studies recommend depletion of DMSO before cell infusion. In this paper, we will briefly review the history of HSC cryopreservation, the side effects reported after transplantation, along with advances in strategies for reducing the adverse reactions, including methods and devices for removal of DMSO. Strategies to minimize adverse effects include medication before and after transplantation, optimizing the infusion procedure, reducing the DMSO concentration or using alternative CPAs for cryopreservation and removing DMSO before infusion. For DMSO removal, besides the traditional and widely applied method of centrifugation, new approaches have been explored in the past decade, such as filtration by spinning membrane, stepwise dilution-centrifugation using rotating syringe, diffusion-based DMSO extraction in microfluidic channels, dialysis and dilution-filtration through hollow-fiber dialyzers and some instruments (CytoMate, Sepax S-100, Cobe 2991, microfluidic channels, dilution-filtration system, etc.) as well. However, challenges still remain: development of the optimal (fast, safe, simple, automated, controllable, effective and low cost) methods and devices for CPA removal with minimum cell loss and damage remains an unfilled need.

Efficient extraction of xylan from delignified corn stover using dimethyl sulfoxide.

Xylan can be extracted from biomass using either alkali (KOH or NaOH) or dimethyl sulfoxide (DMSO); however, DMSO extraction is the only method that produces a water-soluble xylan. In this study, DMSO extraction of corn stover was studied at different temperatures with the objective of finding a faster, more efficient extraction method. The temperature and time of extraction were compared followed by a basic structural analysis to ensure that no significant structural changes occurred under different temperatures. The resulting data showed that heating to 70 °C during extraction can give a yield comparable to room temperature extraction while reducing the extraction time by ~90 %. This method of heating was shown to be the most efficient method currently available and was shown to retain the important structural characteristics of xylan extracted with DMSO at room temperature.

A Pretreatment Method for Analysing Albendazole by HPLC in Plant Material.

Albendazole (ALB) belongs to a group of benzimidazoles—classified as antiparasitic pharmaceuticals. Its widespread application results in the presence of this pharmaceutical in natural environment (water and soil). In this paper a suitable pretreatment method was established including sampling, freeze-drying and extraction. Vicia faba was used as model organism. ALB accumulation by plant tissues was observed in hydroponic culture as well as in soil. The range of pharmaceutical concentrations was 1.7 × 10−5 mol/L (in hydroponic culture) and 1.7 × 10−5 to 1.7 × 10−4 mol/kg air dry soil (in soil). Observations were conducted for 14 days. After this time biological material was freeze-dried and after homogenization, dimethyl sulfoxide (DMSO) extraction was performed. The recovery of ALB for the roots was 93 % while for the shoots 86 %. After cleaning, the samples were subjected to further analysis by HPLC system. Phosphate buffer and acetonitrile (50:50) were used as a mobile phase. Drug retention time was 6.3 min. Results obtained in this experiment indicate higher drug accumulation in roots rather than in the hypocotyl part of the plant, cultivated both in soil and in hydroponic culture.

Amylopectin-like Starch is Present in the Cell Walls of Kiwifruit Pericarp Tissues during Development

The contents of cell wall polysaccharides in pericarp tissues of immature Hayward kiwifruit were determined during development. Two methods employed to remove starch from the crude cell wall preparations were enzymatic digestion (porcine pancreatic -amylase and Pseudomonas iso-amylase, Method 1) and dimethyl sulfoxide extraction (DMSO, Method 2). Method 2 was more effective in removing starch than Method 1. After extensive removal of starch, cell wall materials were successively extracted with 50 mM EDTA, followed by 4% and 24% KOH. The hot EDTAsoluble (pectic) and alkaline-soluble (hemicellulosic I and -II) polysaccharides were further treated with enzymatic digestion. In subsequent amylase digestions of fractions derived from cell walls previously subjected to the two methods the glucose content was further depleted by 28-77% (Method 1) and 85-91% (Method 2) in the pectin fraction, and 51- 86% (Method 1) and 45-91% (Method 2) in the hemicellulose fractions, suggesting starch in the crude cell walls was not released by initial enzymatic treatment or DMSO extraction. Moreover, the crude cell walls yielded -amylase activity upon extraction by 1 M NaCl, further suggesting that starch is present in the cell walls. Methylation analysis of the starch fraction revealed a molar ratio of 0.2~0.6:3.8~11.6:1 for T-Glc, 4-Glc and 4,6-Glc. The red-violet starch-iodine complex had an absorption maximum of 550~565 nm. These results provide evidence for an amylopectin-type starch present in the cell walls of immature kiwifruit. Keywords: Actinidia deliciosa, cell wall, amylopectin, starch, pectin, hemicellulose, xyloglucan, polysaccharides, apoplast, methylation analysis, kiwifruit.

제주산 우뭇가사리(Gelidium amansii)로부터 DMSO 추출과 EDTA 수세법에 의한 상용화 아가로스 제조

Agar was prepared from Gelidium amansii collected from Jeju Island, South Korea. This agar preparation has high gel strength and low sulfate content compared with G. amansii agar from Morocco. Accordingly, agarose was made from the Jeju agar through the consecutive refining processes of dimethyl sulfoxide (DMSO) extraction and ethylene diamine tetra acetic acid (EDTA) washing. The physicochemical properties of the resulting agarose were compared with those from agarose prepared using only DMSO extraction. Consecutive DMSO extraction and EDTA washing more strongly affected the physicochemical properties of the agarose (purified agarose) compared with the use of DMSO extraction alone. These properties were similar to those of commercial agarose used for electrophoresis. In DNA electrophoresis, the separation and movement speed of the purified agarose were similar to those of the commercial agarose. In a <TEX>$^{13}C$</TEX> NMR analysis, the purified agarose exhibited the same carbon peak as the commercial agarose. When observed under scanning electron microscopy, the agar had an even and smooth surface without irregularities or pores, and the purified agarose had a wide surface area with a large number of pores; the commercial agarose had an irregular surface that would allow the solvent to easily permeate. These results illustrate that the physicochemical properties of agarose prepared from DMSO extraction and EDTA washing were more effective than those observed after DMSO extraction alone; thus, these processes used in succession will be useful in agarose industries.

Pulmonary inflammation induced by office dust and the relation to 1 → 3-β-glucan using different extraction techniques

It is observed that 1 → 3-β-glucan, a major cell wall component of fungi, induces pulmonary inflammation. There is inconsistency in determining the correlation between the levels of glucan measured by current extraction methods and the respiratory inflammation observed in individuals or lab animals exposed to environmental dust samples. The glucan-specific limulus amebocyte lysate (G-LAL) method was used after extraction with dimethyl sulfoxide (DMSO) or sodium hydroxide (NaOH) to analyze the glucan content of office dust samples collected from a water-damaged building. C3HeB/FeJ mice, an endotoxin-sensitive strain, were treated with different dust samples (2.5 mg kg−1 body weight) or saline (vehicle control) by pharyngeal aspiration. At 1 day after aspiration, bronchoalveolar lavage (BAL) was performed, and lung inflammation and injury were assessed by measuring: (1) neutrophil (PMN) infiltration, (2) inflammatory cytokine (IL-6, IL-10, MCP-1, IFN-γ, TNF-α, and IL12-p70) levels, and (3) albumin and lactate dehydrogenase in recovered BAL fluid. Both DMSO and NaOH extraction increased the detection of glucan by approximately 20-fold compared to water extraction. However, only the DMSO extraction method showed a statistically significant positive correlation between 1 → 3-β-glucan and albumin levels, total numbers of BAL, polymorphonuclear leukocytes (PMNs) cells recovered, levels of TNF-α, MCP-1, and IL-6. In conclusion, 1 → 3-β-glucan is a potent inflammatory agent in dust samples and DMSO extraction for glucan analysis may prove useful in understanding the impact of environmental contamination by glucans on lung disease.