Hot Water Extraction Method Research Articles

Chemotaxis Assay for Marsupenaeus japonicas Hemocytes and Application for the Development of an Oral Immunostimulant Against White Spot Syndrome Virus.

The chemotactic activity of vertebrate leukocytes is an important host-defense mechanism. However, chemotaxis of invertebrate immune cells, particularly those of shrimp species, is incompletely understood and critically understudied. In this study, we aimed to optimize the conditions for a Boyden chamber chemotaxis assay using hemocytes (granulocytes) from cultured kuruma shrimp, Marsupenaeus japonicas (Mj) and the optimal conditions were: 5 μm-pore-size Polyvinylpyrrolidone membrane; culture buffer at pH 7.0; and chemotactic factor N-formyl-methionyl-leucyl-phenylalanine (fMLP) 10–8 mol/L; 4 h incubation time. We then applied the chemotaxis assay to develop an oral immunostimulant against white spot syndrome virus (WSSV), which results in high mortality rates in several shrimp species worldwide. We focused on the kelp Laminaria japonica, as this species contains immunostimulative molecules such as β-glucan. We prepared Heat Extracts (HE) and Crude Laminarans (CL) from kelp using hot water and hydrochloric acid extraction methods, respectively. HE and CL ware mixed with normal shrimp feed. Kelp extracts were orally administered for 7 days, and hematocyte chemotaxis toward fMLP was compared. No difference was detected between control and kelp extracts on day 3, but HE stimulated chemotaxis 2-fold and CL stimulated chemotaxis 3-fold relative to control on day 7 after initiating administration. Kelp extract administration protected against WSSV exposure. Finally, we identified that Kelp extracts stimulated hematocyte superoxide production on days 3 and 7, and increased hematocyte phagocytosis and phenol oxidase activity on day 7 after administration. We concluded that the chemotaxis assay is informative in assessment of shrimp hemocyte immunological activity, and is applicable to the development of immunostimulants against shrimp infectious diseases.

Structural features, selenization modification, antioxidant and anti-tumor effects of polysaccharides from alfalfa roots

Hot water extraction and chromatographic purification methods were used to extract and purify two polysaccharides (RAPS-1 and RAPS-2) from the roots of alfalfa. Subsequently, RAPS-2 was modified using the HNO3/Na2SeO3 method to obtain Se-RAPS-2. The structural features, antioxidant and in vitro anti-tumor activities of the three polysaccharides were evaluated. The structural analysis revealed that RAPS-1 (Mw = 10.0 kDa) was composed of rhamnose, xylose, arabinose, galacturonic acid, mannose and glucose, whereas RAPS-2 (Mw = 15.8 kDa) consisted of rhamnose, xylose, galacturonic acid, mannose, glucose and galactose. RAPS-1 contained 1 → 2, 1 → 4, 1 → 3, and 1 → 6 or 1 → glycosidic bonds; however, while RAPS-2 lacked 1 → 4 glycosidic linkages. The molecular weight of Se-RAPS-2 was 11.0 kDa less than that of RAPS-2. The results of activities demonstrated that Se-RAPS-2 displayed superior antioxidant activity and inhibitory effect in HepG2 cells than RAPS-1 and RAPS-2.

In Vitro Evaluation of The Antioxidant and Anti-Skin Aging Properties of Green Algal Sulfated Polysaccharides

Abstract Skin aging is a natural phenomenon witnessed by humans. However various intrinsic and extrinsic factors lead to early skin aging. There have been a variety of approaches to combat skin aging one approach uses antioxidants that are known to fight oxidative stress as well as combat problems of aging. In this study, the antioxidant and anti-skin aging properties of sulfated polysaccharides (SPs) from fresh water microalgae Chlamydomonas reinhardtii (Cr) are evaluated. Sulfated polysaccharides were isolated by hot water extraction method and purified by anion-exchange chromatography. The biochemical composition of the extract showed carbohydrate content of 785.07 mg/g, 324.26 mg/g of sulphate and 393.32 mg/g of uronic acid. These extracts which are enriched with SPs were further used for checking antioxidant and anti-skin aging properties. Cr-SPs showed Superoxide anion scavenging activity of 38-92% at 0.1-2 mg/mL, 51-89% of nitric oxide scavenging ability at 0.2-2 mg/mL, 10-58% of hydrogen peroxide scavenging ability at 1- 10 mg/mL, and 28-68% of ferric ion reducing potential at 0.5-5 mg/mL respectively. Furthermore, Cr-SPs showed 90% anti-elastase enzyme activity at 1 mg/mL, 83% and 89% anti-collagenase and anti-hyaluronidase activities at 1 mg/mL and 2 mg/mL respectively. These promising antioxidant and anti- skin aging properties of Cr-SPs pave way to explore the potential of Cr-SPs in cosmeceutic and pharmaceutical formulations as anti-skin aging agents in a cost-effective manner.

The antioxidant activities of garlic polysaccharide and its derivatives

Garlic was used as raw material to extract garlic polysaccharide (P) with hot water extraction method. IR and NMR spectra showed that the preparation of sulfated garlic polysaccharide (SP), phosphorylated garlic polysaccharide (PP) and carboxymethylated garlic polysaccharide (CMP) was successful. The sugar content of P, SP, PP and CMP was determined, and their degree of substitution was measured. Hydroxyl radical scavenging ability, superoxide anion scavenging ability, significant reduction capacity, and anti-lipid peroxidation ability of P, SP, PP, CMP and VC were determined. It showed that the introduction of substituents had different effects on the antioxidant activities of P.

Preparation, deproteinization, characterization, and antioxidant activity of polysaccharide from Medemia argun fruit

In this study, the crude polysaccharide from argun palm (Medemia argun) fruit was extracted with hot water and deproteinized by four different methods. Its structure characterization, thermal property and antioxidant activity were investigated by FT-IR spectroscopy, Scanning electron microscope, Congo-red, I2-KI, DPPH• assay, ABTS•+ assay, TGA and DSC analysis. It indicated that the NaCl method was the best method in deproteinization. The argun fruit polysaccharide linked by the β-glycosidic bonds. The molecular weight of polysaccharide deproteinized with NaCl was 4.46 × 105 Da. It showed that the polysaccharide from argun palm fruit consisted of fructose, arabinose, rhamnose, galactose, glucose, xylose, and galacturonic acid. The result of TGA and DSC assay indicated that argun fruit polysaccharide has a favorable thermal stability. It was proved that, the argun fruit polysaccharide possessed high antioxidant activity and would be a new natural antioxidant.

Comparison of different extraction methods of polysaccharides from cup plant (Silphium perfoliatum L.)

The purpose of the present research was to compare the effects of four extraction methods on the extraction yields, physicochemical characteristics, and antioxidant activities of polysaccharides from the cup plant (Silphium perfoliatum L.) (CPPs). The extraction methods included the hot water extraction method (HEM), ultrasonic-assisted extraction method (UEM), enzyme-assisted extraction method (EEM), and the ultrasonic-enzyme-assisted extraction method (UEEM). Four corresponding CPP extracts, HEM-CPPs, UEM-CPPs, EEM-CPPs, and UEEM-CPPs, were obtained. The results indicated that there were notable differences between the extraction methods regarding the yield, average molecular weight, uronic acid content, monosaccharide proportions, and the antioxidant activities of the CPPs. HEM-CPPs were extracted with the lowest yield (6.44%). EEM-CPPs had the largest extraction yield (9.87%) and the most notable antioxidant abilities of ferric reducing power and free radical scavenging activity. According to correlation analysis, the higher antioxidant abilities of EEM-CPPs might be due to their smaller average molecular weight and a higher content of uronic acid than those of the other extracted CPPs. Therefore, EEM could serve as a promising technique for extracting CPPs.

Development, chemometric-assisted optimization and in-house validation of a modified pressurized hot water extraction methodology for multi-mycotoxins in maize

Effective management of mycotoxins rely on stringent regulation and routine surveillance of food/feed commodities via efficient analysis, hence the continuous need for improved methods. The present study developed, optimized and validated a modified pressurized hot water extraction (PHWE) method for the simultaneous extraction of multi-mycotoxins from maize and subsequent quantification on LC-MS/MS. The PHWE system was modified using ethanol (EtOH) as a cosolvent, while a numerical modelling approach, the central composite design (CCD), was adopted for the optimization of the extraction conditions. Using the optimized method, it was possible to effectively extract and quantify 15 different mycotoxins from maize in a single step with satisfactory linearities (0.986–0.999), recoveries (14–124%) and other associated method validation parameters. Further efficacious application of the method to real samples re-affirmed the prospects of PHWE as a suitable, cost-effective and greener alternative to traditional methods of mycotoxin extraction.

Isolation of crude fucoidan from Sargassum wightii using conventional and ultra-sonication extraction methods

Fucoidan, a fucose rich sulphated polysaccharide, generally found in cell wall matrix of brown seaweeds possess diverse physiological properties. Crude fucoidan was extracted from Sargassum wightii, a brown seaweed common along the Indian coastline. A comparison was made between hot water extraction (HWE) and ultra-sonication extraction (UAE) methods. The UAE method gave higher yield (14.61 g/100 g) than HWE method (10.59 g/100 g). The UAE fucoidan had lighter colour than HWE, as indicated by Hunter colour values. The water activity values for fucoidans obtained were 0.14 for HWE and 0.19 for UAE. The sulphate content was similar in both fucoidans (̴17%). However, HWE fucoidan had higher content of fucose. The Fourier transform infrared spectroscopy spectra confirmed the presence of sulphate groups in both the fucoidans. The absorption bands at 1245.37 cm−1 and 1294.99 cm−1 in the HWE and UAE fucoidan respectively, indicated the S–O stretching due to sulphate groups in the fucose monomer. The thermal transition temperature as obtained by differential scanning calorimetry for HWE (127.3 °C) was lower than UAE fucoidan (131.8 °C). HWE fucoidan had higher DPPH radical scavenging and reducing power than UAE fucoidan. On the other hand, UAE fucoidan had higher metal chelating activity. The present study demonstrated that ultra-sonication can be an effective alternative for the extraction of sulphated polysaccharide from seaweed, in term of higher yield. The health benefits of Sargassum wightii and fucoidan obtained from it, could be explored by incorporation in different food systems.

Flaxseed gum: Extraction, bioactive composition, structural characterization, and its potential antioxidant activity.

Flaxseed gum (FSG) is a heteropolysaccharide consist of neutral and acidic components that makes up approximately 8% of seed mass. FSG was extracted from mixture of brown and golden varieties of flaxseeds by hot water extraction method. The molecular weight distribution, monosaccharide analysis, chemical composition, and surface morphology of FSG were scrutinized in the current study to get a better insight regarding this important polysaccharide. The average molecular weight was recorded as 1,322kDa with a polydispersity ratio of 1.6 for Mw/Mn and 2.4 for Mn/Mz. High-performance liquid chromatography (HPLC) screening revealed that extracted FSG was comprised of rhamnose, arabinose, mannose, glucose, fucose, xylose, galactose, glucosamine, glucuronic acid, and galacturonic acid, of which mannose and glucosamine have not been reported previously. The antioxidant activities of FSG measured as DPPH, ABTS, reducing power, and total antioxidant activity clearly demonstrated the antioxidant potency of FSG. Fourier-transform infrared spectroscopy (FTIR) and the Nuclear magnetic resonance (NMR) (1 H, 13 C) s pectra ratified the presence of functional groups typical for polysaccharide. PRACTICAL APPLICATIONS: Flaxseed gum is a natural carbohydrate polymar. This study provides useful information regarding antioxidant potential, chemical, and structural characterization of FSG. The availability of a gum is fully characterized with bioactive composition, structural features, and antioxidant potential provides a toolset for the practical application in the food or drug industry.

In vitro anti-HIV-1 activity of the bioactive compound extracted and purified from two different marine macroalgae (seaweeds) (Dictyota bartayesiana J.V.Lamouroux\xa0and Turbinaria decurrens\xa0Bory)

Highly active antiretroviral therapy (HAART) is the only available remedial measure to treat HIV infected patients, as recognized by the WHO. However, it is associated with toxicity (nephrotoxicity), high cost and most preferably drug resistance in the first-line treatment. Wherefore, potential and novel natural source is the only option for the modern world to challenge this global issue. In recent years, sulfated polysaccharide from marine macroalgae shown to be biologically active as anti-inflammatory, anticoagulant, antitumor, immunomodulatory and antiviral agents. As a direct inhibitor of HIV including other retroviruses, it is considered as a “new generation antiretroviral drug”. In our present study, Fucoidan, a sulfated polysaccharide has been extracted from two different macroalgae Dictyota bartayesiana (DD) and Turbinaria decurrens (TD) based on hot water extraction method and further confirmed by FT-IR and RP-HPLC methods. Both the crude and purified fucoidan samples were evaluated for anti-HIV activity after ion exchange chromatography purification. The maximum inhibitory activity of crude and purified fucoidan samples are 90.5% and 89% in the fucoidan extracts of DD. Whereas, it was 89.7% and 92% in the fucoidan extracts of TD. Simultaneously, the IC50 values were determined and recorded as 1.56 µg/ml and 57.6 ng/ml in both the crude and purified fucoidan extracts of DD respectively. Similarly, for TD, it was 3 µg/ml and 131.7 ng/ml in the fucoidan extracts of TD. Therefore, further extensive research work is the most needful to fill the gaps to develop this sulfated polysaccharide as a potential drug for the treatment of HIV patients.

The Effect of Extraction Methods on Preliminary Structural Properties and Antioxidant Activities of Polysaccharides from Lactarius vividus

Four polysaccharides (LVP-u, LVP-m, LVP-e, and LVP-h) were extracted from the fruiting bodies of Lactarius vividus by an ultrasonic-assisted extraction method, microwave-assisted extraction method, enzyme-assisted extraction method, and hot water extraction method, respectively. Then, the effect of extraction methods on yields, preliminary structural properties, and antioxidant activities was systematically investigated using the weighing method, chemical composition analysis, high-performance gel permeation chromatography (HPGPC), high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), radical scavenging, and metal ion chelating assays. Results demonstrated that the four L. vividus polysaccharides (LVPs) were all combined with protein, and the yield of LVP-u was higher than others. Molecular weight distribution, monosaccharide and amino acid compositions, and microstructures among the four LVPs were significantly different. Moreover, the LVPs showed significant antioxidant activities in a dose-dependent manner, and LVP-e demonstrated better antioxidant activities in superoxide anion radical scavenging activity assays and metal ion chelating activity assays, while LVP-u showed higher activity in its hydroxyl radical scavenging ability.

Extraction of polysaccharides from Amaranthus hybridus L. by hot water and analysis of their antioxidant activity.

BackgroundAmaranthus hybridus L. is an annual herb that belongs to the Amaranthceae family, a type of multi-purpose grain, vegetable and feed crop that has received considerable attention due to its great economic value. However, the composition of polysaccharides from A. hybridus has rarely been previously reported.MethodsIn this study, the aboveground part of A. hybridus was used as material and polysaccharides were isolated by the hot water extraction method. Two acidic polysaccharides were isolated and purified by the Sevage method and diethylaminoethyl cellulose-32 column chromatography.ResultsTwo acidic polysaccharides were obtained from A. hybridus: AHP-H-1 and AHP-H-2. There were significant differences between the monosaccharide content from each sample according to gas chromatography-mass spectrometer. AHP-H-2 had higher antioxidant activity in vitro than AHP-H-1. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging rate of two mg/mL AHP-H-2 was 80%, its hydroxyl radical scavenging rate was approximately 48.5%, its superoxide anion radical scavenging rate was 85.3% and its reduction ability of Fe3+ was approximately 0.92. The total antioxidant capacity of each milligram of AHP-H-2 was 6.5, which was higher than ascorbic acid.ConclusionThe results of the study promote the effective use of A. hybridus and provide a theoretical basis for its development.

Structural characterization of a polysaccharide from Certaria islandica and assessment of immunostimulatory activity

Abstract In this study, Cetraria islandica polysaccharide (CIPs) was extracted by hot water extraction method and its effect on structural modification of immunomodulatory activities was investigated. This polysaccharide mainly consisted of carbohydrates (97.0%), Sulfate (1.2%) with one type of glucose. The average molecular weight (328.7 × 103 g/mol) was determined by size exclusion chromatography. The structure of the polysaccharide was found to be composed of (1→3) and (1→4)-β-D-glucopyranosyl units. The immunomodulatory activities of the crude polysaccharide and its derivatives, over-sulfated (OS1,2,3) and hydrolyzed (H1,2,3), were tested against NK-92 cells and RAW264.7 cells. The results obtained clearly demonstrated that over-sulfated (OS1,2,3)-treated NK-92 cells induced cytotoxicity in HeLa cells through the expressions of IFN-γ, NKp44, NKp30, and FasL. On the other hand, the hydrolyzed derivatives (H1,2,3) activated RAW264.7 cells through production of nitric oxide (NO) and mRNA expression of iNOS, IL-1, TNF-α, IL-6, IL-10 and IL-12 through the nuclear factor kappa-B (NF-κB) and mitogen activated protein kinases (MAPK) pathways. Our findings suggest that the presence of sulfate in the polysaccharide played a pivotal role in NK-92 cell however the Mw was a determinant factor in RAW264.7 cell activation, in which both cells were activated through the CR3 and TLR-4 signaling pathways.

Inulin/PVA biomaterials using thiamine as an alternative plasticizer

In this study, biodegradable biomaterials were prepared by using inulin (INL), PVA and plasticizers (citric acid (CA), glycerol (GL) and thiamine (TH)) with UV curing process. INL was extracted from Jerusalem artichoke flour using hot water extraction method. Extracted INL and INL/PVA biomaterials were characterized by TLC, FT-IR, and SEM analysis. Physical properties such as mechanical and water resistance properties of biomaterials prepared with UV curing time from 0 to 20 min and types of plasticizers were investigated. Their antimicrobial activities, biodegradability, and application of coating materials for foods were also determined. Results indicated that their physical properties were improved by the UV curing process. In addition, physical properties of TH-added biomaterials were 1.5 to 2 times higher than those of GL-added and CA-added biomaterials. Biodegradability in soil revealed that biomaterials were degraded by about 20–40% after 140 days.

Polysaccharides from Trifolium repens L. extracted by different methods and extraction condition optimization

Four different extraction methods, including hot water extraction (HWE), ultrasonic-assisted extraction (UAE), enzyme-assisted extraction (EAE) and ultrasonic-enzyme-assisted extraction (UEAE), were applied to extract polysaccharides from Trifolium repens L. (TRPs). In addition, response surface methodology (RSM) was performed to optimize the extraction conditions of TRPs. The results showed that different extraction methods had significant effects on the extraction yields and antioxidant activities of TRPs. TRPs extracted by the EAE method (10.57%) and UEAE method (10.62%) had significantly higher extraction yields than TRPs extracted by the HWE method (8.35%) and UAE method (9.43%) (P < 0.05). However, there was no significant difference between the extraction yields of the EAE method and UEAE method (P > 0.05). TRPs extracted by the EAE method had a higher content of uronic acid and exhibited better antioxidant capacities. Therefore, EAE was selected as the optimal extraction method to extract TRPs. The optimal extraction conditions of EAE to extract TPRs were liquid–solid ratio 30 mL/g, enzymolysis time 87 min, enzyme-complex dosage 1.6% and pH 6, leading to a TRPs yield of 13.15%.

Characterization of Moringa oleifera roots polysaccharide MRP-1 with anti-inflammatory effect.

Moringa oleifera is a mutli-purpose herbal plant which has attained enormous attention as a natural source of nutrients and folk medicine. This work aimed to get a novel polysaccharide, termed MRP-1, which was isolated from Moringa oleifera roots with hot water extraction method followed by ethanol precipitation and purified with DEAE-Sepharose Fast Flow column. Monosaccharide composition analysis based on GC-MS showed that MRP-1 mainly consisted of Rha, Ara, Fru, Xyl, Man and Gal in the molar ratio of 1.5:2.0:3.1:6.0:5.3:1.1. The Roman spectra, FT-IR and NMR analysis showed that the typical features of carbohydrates, such as α-Araf, α-Gly, β-Galp, α-GalpA and β-Gly was contained by MRP-1. The LPS-induced RAW264.7 macrophage cells were used to evaluate the anti-inflammatory activity of MRP-1. The result demonstrated that the increasing of NO and TNF-α production induced by LPS could be prevented by different concentrations of MRP-1 treatment. Moreover, the mRNA expression level of iNOS induced by LPS was decreased significantly (p < 0.05) by MRP-1 treatment while show no obvious effect on the COX-2 mRNA expression. This study may provide new possible application of Moringa oleifera root polysaccharide related to anti-inflammation.

Optimization, Characterization and Antioxidant Activities of Selenized Polysaccharides from Hypsizygus marmoreus

In this study, hypsizygus marmoreus was used as raw material to extract polysaccharides by hot water extraction method. Preparation technology of Selenized Polysaccharides from Hypsizygus marmoreus (SPHM) and its antioxidant activities were investigated. Chelation rate was selected as the index, effects of initial concentration of sodium selenite (mg/mL), the mass ratio of polysaccharide to sodium selenite, chelation time and pH on the chelation rate was investigated by single factor test. Response surface method was used to optimize the preparation technology of SPHM. The preliminary structure characterization of Polysaccharides from Hypsizygus marmoreus (PHM) and SPHM was performed by infrared spectroscopy and scanning electron microscopy. The antioxidant activities of SPHM were studied by the scavenging effect of 1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1- (2,4,6-trinitrophenyl) Hydrazyl (DPPH) and hydroxyl radical. The results showed that the optimum preparation technology of SPHM was as follows: the initial concentration of sodium selenite was 7.95 (mg/mL), the mass ratio of polysaccharide to sodium selenite was 6.11:1 (mg/mg), chelation time was 6.89h and pH was 5.11. Under the optimum conditions, chelate rate was 10.36%. Infrared spectroscopy and scanning electron microscopy analysis showed that selenium ions successfully combined with PHM and the surface structure of PHM and SPHM was different. Compared with HPM, the antioxidant activities of SPHM enhanced.

Fructus mori L. polysaccharide-iron chelates formed by self-embedding with iron(iii) as the core exhibit good antioxidant activity.

Mulberry fruit polysaccharide (MFP) was obtained from Morus alba L. by a hot water extraction method, and mulberry polysaccharide fractions named MFP1, MFP2 and MFP3 were isolated by DEAE cellulose-52 column chromatography. Monosaccharide analysis of MFP1, MFP2 and MFP3 showed that the three components had the same monosaccharide compositions with different ratios, and galacturonic acid was the main monosaccharide component. Molecular weight measurements showed that MFP1 and MFP2 are heteropolysaccharides and MFP3 is a homogeneous polysaccharide. In addition, the chelate mechanism of iron(iii) and polysaccharide is proposed in which iron(iii) as a core is enwrapped by the polysaccharide as a ligand by hydroxyl and carboxyl groups, which induces a morphology change from flat sheets to rods and increases the size. Furthermore, the polysaccharides showed strong antioxidant activity to eliminate hydroxyl radicals and inhibit MDA production in healthy mouse liver homogenate. Also, the polysaccharide-iron(iii) chelates exhibited stronger superoxide radical scavenging ability than the polysaccharides. These results suggest that the polysaccharides derived from Morus alba L. are promising candidates for fabricating organic iron supplements with good antioxidant activity.

Antioxidant activities of sulfated pumpkin polysaccharides

Pumpkin polysaccharide was extracted by hot water extraction method. It was modified with chlorosulfonic acid-pyridine to obtain sulfated pumpkin polysaccharides (SP1, SP2) with different degrees of substitution, which was 0.35 and 0.65, respectively. The total sugar contents of pumpkin polysaccharide and its sulfated derivatives were determined by phenol‑sulfuric acid method, the ability of scavenging hydroxyl radicals and superoxide anions as well as the ability of reducing were determined. The results showed the scavenging effect of sulfated derivatives on hydroxyl radicals was not different after 0.8 mg/mL and was lower than that of pumpkin polysaccharides. The sulfated pumpkin polysaccharides with different degrees of substitution had better scavenging effects on superoxide anions than pumpkin polysaccharide, but the reducing ability was lower than that of pumpkin polysaccharide.

Different Extraction Techniques on the Characteristic Properties of Valonea Tannin

Valonea extract is a natural tannage widely applied in leather industry which is produced via conventional hot water extraction method. On the otherhand more environmental friendly methods have started to be used in extraction technology recently. Present paper reports on the effect of different extraction methods on the properties of valonea extracts. For this purpose, conventional solvent extraction (CSE) (methanol/water mixture), supercritical CO2 (scCO2) extraction and pressurized fluid extraction (PFE) (water/CO2 binary system) techniques have been employed on valonea and the obtained extracts have been characterized. Molecular weight of the tannins was determined by gel permeation chromatography (GPC), particle size was examined by Malvern Zetasizer instrument, chemical structure by Fourier Transform Infrared Spectroscopy (FTIR). Tannin content was also examined and the results were compared with commercial valonea extract (CVT). The results showed that especially particle size of the tannins was affected by the applied extraction method and the smallest particle size was obtained by PFE method as 505.3 nm compared to commercial tannin as 983.0 nm of average particle size. PFE method was also provided more uniform extract according to the polydispersity index values. GPC and FTIR analyses showed slightly differences on the characteristics of tannins.