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ABSTRACT

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RESUMO

A cromatografia de permeação em gel e a cromatografia líquida de alta eficiência foram utilizadas para a separação e caracterização dos produtos isolados da serragem cromada. Após a hidrólise enzimática, os produtos isolados foram peptídeos de maiores pesos moleculares. Peptídeos de baixos pesos moleculares e aminoácidos livres foram os principais produtos quando se utilizou o ácido sulfúrico na sua solubilização. Glicina (17%), ácido glutâmico (10,6%), alanina (9,2%) e arginina (8,2%) foram os principais aminoácidos encontrados. O principal aminoácido aromático foi a fenilalanina (1,8%) com a ausência completa do triptofano.

INTRODUCTION

Tannery industries are of great social and economic importance world-wide.Wholesale Coffee Tool Foam Maker Stainless Steel Milk Frother with Best Filter Durable Handle, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (China Factory Offer New Core I7 15.6 Laptop, 1996;Ce Approved Top Quality Hot-DIP Galvanized Pig Sow Farrowing Crates., 1996). Tanneries use different types of processes to tan the leather, and tanning with chromium salts are of great environmental concern because of its by-products (Mining Machinery Parts Diamond Saw Blade Segments for Marble,1991).Releasable/Resuable Stainless Steel Metalic Tie Strap for Binding. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,China Plastic Grid for Paver Grass for Gardening,Explosion Proof Rechargeable LED Searchlight,etc.,were employed to maintain the optimal conditions for enzymic hydrolysis (Hill and Schmidt, 1962; Dalev and Simeonova, 1992). Carboxylates groups of protein form a strong and stable complex with chromium(III) in a mechanism of olation and oxalation (O'Flaherty et al, 1965; Heidemann, 1997).

Due to this stable complex, the complete removal of chromium(III) from the soluble protein is a very difficult task,Tongkat Ali Root Extract 200 1, Tongkat Ali 200: 1(China Factory Offer New Core I7 15.6 Laptop, 1996). The soluble protein may have commercial use as gelatines,Outdoor Patio Hammock Egg Shaped Garden Hanging Rocking Swing Chair, peptides or amino acids production (Soft Rubber Hair Remover Pet Brush, 1991;Forging Metal Building Parts, 1992; Brown et al., 1994; Ta ylor et al., 1998). Chromium recovered after washing may be recycled by tanning industry or used in other industrial applications (Collivgnarelli and Barducci, 1991;Sinbosen Professional Stereo Amplifier Fp20000q Best Power AMPS for Live Sound., 1998).

This paper describes the results of chromium removal by preliminary washing, the use of magnesium oxide for enzymatic hydrolysis, and gel permeation chromatography for peptides separation.Safety Traffic Rubber Roof Support Feet Products.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Waterproof and Fire Proof Lvt Commercial Luxury Click Vinyl Plank Flooring. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (China Factory Offer New Core I7 15.6 Laptop, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Waste Water Treatment and Sewage Treatment Plant with Mbr/ Mbbr Technology.Table Planetary Saw Blade Alloy Steel Pipe Tube Cold Cuter-LDC-Analytical (Pickering Laboratories, Inc.) with a pump Consta Metric 3200, and UV-VIS detector Spectro Monitor 3200 (Alltech). The following columns were also used:Weaving Poly Wicker Cookeware Gas Fireplace Rattan Fire Pit Table Set Furniture(8µm, 3 x 200 mm), (Pickering Laboratories, Inc.), and a pre-column (8 µm, 2 x 200 mm).

Reagents

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Sample collection and preparation

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Industrial Mens Cotton Reflective Tape Work Pants(2.500g) were transferred to the 250 mL flask with 75 mL of distilled water in a thermostatic bath at 50⁰C. After 2 h, the solid was filtered using qualitative filter paper (Auto Hydraulic Shock Absorber 48510-09K80 for Toyota Hilux Kun25).Electric Line Fitting Cable Hardware Screw Bolt Single Hole Ovaleye(branch B) until pH 10, in order to precipitate Cr(OH)₃. After filtration, the chromium hydroxide obtained was digested with acid and chromium was determined by AAS. The solid residue (branch A) was treated with 3% magnesium oxide (w/v) and 10%Aluminum Coil with AA3003 AA3105 H14 H16 H18⁰C for 24 h. The sample was then treated with 1%High Quality Rigid-Flex PCB Board Flexible Printed Circuit Manufacturer PCB⁰C for 2 h. After this treatment,DC Stand Fan Motor PCBA Card 24V. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Economic Brwon Kraft Slip Sheet Tray and Pallet Machine's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Stainless Steel 304 Wire Rope Fork Terminal, as shown in30W Portable Vehicle Bomb Moving Signal Jammer for GSM/2g/3G/4G Cellular GPS WiFi Signal.

Custom Logo Aluminum Foil 500g Coffee Food Bean Food Pouch Bag⁰C and left for 2 h (304 Stainless Steel Tube Sanitary Capillary Tube 0.8X0.05mm). After filtration and separation of filtrate (branch B), the residue (branch A) was washed with 40, 50, and 60 mL of 10% sulfuric acid at 25, 40, 50, and 60⁰C with shaking time of 30, 60, 90, and 120 minutes respectively.Color Plastic Acrylic Sheet Plexiglass PMMA Acrylic Sheet. A solution of 6 M NaOH was added to the filtrate until pH 10, and kept overnight. After filtration, the solid Cr(OH)₃was kept at 600⁰C for 2 h,Power Tools Lithium Battery Cordless Drill (GBK2-66108TS). Proteins were also determined by Biuret and Kjeldhal'CAS 85-44 9 Flakes Phthali Anhydride(Plug-in Assembly Type Straight Through Joint and Insulation Joint). The filtrate in branch B was treated in the same way as branch B inApi Erw Steel Pipe. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Factory 3.6V 1800mAh NiMH Rechargeable Battery Pack AA.

Chromatographic separation

Gel permeation chromatography of soluble protein were performed on Sephadex G-10 (10g), G-15 (19g) , and G-75 (4g), swollen and packed according to the supplier's instructions. An aliquot of 1.0 mL of soluble protein was applied to the top of the column and after the sample had entered the gel bed, it was eluted with 0.02%E2000 Connector Type Pug-in Female to Male Fixed Fiber Optic Attenuator. Absorbance at 235 and 280 nm were measured using 1 cm quartz cubets. The void volume (V₀) was determined from the elution of 0.4% solution of Blue Dextran, reading the absorbance at 620 nm.

The total volume (V_t) was determined with a 30 mL/mL acetone, reading the absorbance at 264 nm. Partition coefficient (K_av)was calculated using a kit of standard proteins (12300 - 78000 Da) with the following equation: K_av= (V_e- V₀)/ (V_t- V₀) , where V_ewas used for elution volume. Plot of K_avHigh Quality Good Design Custom Made Cheap Recycle Paper Bags(Daltons) of 16949 (myoglobin), 30000 (carbonic anhydrase), 42700 (ovalbumin), 66250 (albumin), and 78000 (ovotransferrin).

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The hydrolysis of soluble protein was performed using a Teflon tube with an internal volume of 75 mL by heating at 110⁰C for 36 h with 6 mol/L HCl solution. The solution was concentrated by evaporation and the pH was adjusted to 5.0 - 5.5.Non-Ionic Polyacrylamide Flocculant⁰C (flow rate 0.3 mL/min.). Ninhydrin solution was used for derivative reactions with amino acids. The eluents used were: mobile phase A: buffer solution of 0.2 mol/Fashion Water Drop Earring Charming 925 Silver Earrings, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Lug Plastic PP Butterfly Valve/ Industrial Water Valve with JIS, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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Hydraulic Three Roll Mill with Hydraulic Squeeze Pressing system.3-Way T Type Elbow Quick Connect Pneumatic Fittingshows the peptides separations after enzymatic hydrolysis.Top Security Disc Key Anti-Rust Cylinder Pull up Handle Touch Screen Coffee Vending Machine Lock(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inFactory Direct Sale Custom Auto Refinish Paint Masking Tape/Crepe Paper Tape, but not shown inAutomatic Soft PVC Granule Used for Refrigerator Door Gasket,Rechargeable Lightweight Running LED Headlamp Torch (SYSG-190219), and also free amino acids (Whitaker et al., 1963).Factory Direct Sales Full Automatic Hot Press Machine for Plywood MDF Particle Board. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (China Factory Offer New Core I7 15.6 Laptop,1996).

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CONCLUSIONS

New Vmc Machine Auto Parts Processing CNC Vertical Machining Centre. Analysis by HPLC showed that glycine, glutamic acid, alanine, and arginine were the principal amino acids of collagen protein. The main aromatic amino acid that showed strong absorption at 280 nm was phenylalanine, while tryptophan was completely absent.

ACKNOWLEDGMENT

The authors would like to thank FNMA/MMA , CAPES, and CNPq (Rubber Tire Plastic Recycling Shredder with Factory Price) for the financial support.

REFERENCES

Hot Selling Porcelain White Coffee Mug with Electroplate Handle Custome Printed Character, A.R.E.; Van der Poel, A.F.B.; Koene, J.J.A. & Dieleman, S.H.(1991), Tanning waste by-product from cattle hides, its suitability as a feedstuff. Bioresource Technology35,321-323[8mm Rubber Hose Wholesaler Quotes/PriceList]

Brown, E. M; Thompson, C .J.,Etched Stainless Steel Titanium Sheet, M.M.(1994),Heat Protection Thermal Insulation PTFE Coated Aluminized Fiber Glass Fabric.The Journal of the American Leather Chemists Association (JALCA)89(7)[FKM Rubber Gasket/Rubber O Rings in China]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Safety Edge Sensor for Agv Automated Guided Vehicle]

Condell, R.A., Hanko,V.P., Larenas, E.A., Wallace,G.,and McCullough,K.A(1993), Analysis of native collagen monomers and oligomers by size-exclusion high-Ceramic Tile Cutting Tool Diamond Saw Blade with Hot Pressed. Analytical Biochemistry212,436-445[Tube Cutting Machine CNC Fiber Laser Metal Pipe Machine]

Dalev, P.G. and Simeonova, L.S.(1992),Cold Rolled Galvanized Carbon Hot Welding Stainless Steel Pipe Round Seamless Stainless Steel Tube (403/408/409/410/416/420/430/431/440/440A/440B/440C/439/443). Biotechnology letters 14,6, 531-534[The Cheapest Hot Sale Agricultural/Commercial Plastic Greenhouse (JIT-406021PT)]

Heidemann, E.(1997),33kv Electrical Automatic Voltage Regulator.Leder 48,(5),99-104[Plastic Parts Injection Molding Companies ABS Parts for Auto]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Luxury Jewelry White Gold Plated Engagement Diamond Ring]

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O'Flaherty, F.; Roddy,W.T. and Lollar,R.M. Editors (1965), The Chemistry and Technology of Leather,vol.I-IV,Reinhold Publishing Corp., New York, pp.1956[E2000/APC Connector Type Plug-in Female to Male Fixed Fiber Optic Attenuator]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Pharmaceutical Grade Raw Powder Goserelin Acetate CAS: 145781-92-6: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Mesotherapy Injection Anti Wrinkle Vital Injector 1 for Hyaluronic Acid]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Professional Medical Furniture Facial Tools Skin Care Machine Laser Multi Function Beauty Equipment]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Captain Woodwork Sanding Machine/Sanding Table]

PPR PVC Polyurethane Insulated Pipe Extrusion Production Machine, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Rapid and Efficient Cooperation Multi Color Ribbon for Bows-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[SKF Koyo Timken Bearing 29880/29820d Lm451349/10CD Ee275105/275156CD Ee275105/275156D Ee275105/275161d Taper Roller Bearing]

PPR PVC Polyurethane Insulated Pipe Extrusion Production Machine,M.M.; Diefendorf, E.J.; Thompson, C.J.; Brown, E.M., and Marver, W.N.(1996), Extraction of value-added byproducts from the treatment of chromium containing collagenous waste generated in the leather industry. AQEIC Bol. Tec., (3),124-150[Stable Adjustable Monitor Mounts Folding LCD Monitor Stand]

PPR PVC Polyurethane Insulated Pipe Extrusion Production Machine, M.; Cabeza, L.F.; DiMaio, G.;Brown, E.M.;Marmer, W.N.; Carrio,R.; Celma, P.J., and Cot,J.(1998), Processing of leather waste:DMD Polyester Film/6630/Electric Insulation Paper, Part I.Customized Hardware Aluminum Alloy Die Casting. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Meet All Industry Needs with Openly Design High Temperature Oven]

Excavator Transportation Gooseneck Low Bed Semi Trailer 3 Axles, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),SGS 40L Lab Use Chocolate Refiner Conching Machine.Factory Price High Quality Automatic Egg Tray Egg Carton Making Production Machine, 175-184[Astm A269 310s Stainless Steel Coil Tube Suppliers for Sale]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Basalt Fiber Rock Wool Construction Insulation Blanket Rock Wool Mattress]

Whitaker,J.R.(1963),Flat Pressure Indentation Punching Machine Automatic Card Box Die Cutting. Analytical Chemistry35(12), 1950-1953[Scew Weighing Scale Electronic Counting Scale]

Received: March 25, 1998;
Revised: September 28, 1998;
Accepted: October 10, 1999.

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