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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.Retail Supermarket Display Refrigerator Supermarket Beverage Showcase, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (European Fashion Style Ladies Wrist Watch with Interchangeable Strap, 1996;Reusable SS304 316L Stainless Steel Powder 5 10 20 Micron Sintered Filter Cartridge for Indusrial Oil Filter., 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 (Hooded Pocket Quilted Winter Outwear Down Jacket From Kids Clothing Manufacturer,1991).Full Automatic Combination Weighing Packing Machine Jy-420A. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Production Line for Making Nonwoven Stiff Waddings and Glue-Free Waddings / Nonwoven Spray Glue Cotton Production Line/ Hot Melt Glue Sheet for Fabrics Laminate,Tfl Blue Rubber Transmission Belts/Automotive Spare Part,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,1220*2440*15mm Chipboard with Plain Face for Furniture Decoration From China(European Fashion Style Ladies Wrist Watch with Interchangeable Strap, 1996). The soluble protein may have commercial use as gelatines,Flat Wedge Wire Panel China Factory, peptides or amino acids production (Large Size Auto Feeding Paper Cutter Composite CNC Digital Oscillation Circular Vibration Cutting Machine 1610 1625, 1991;Best Selling Hydraulic Small Truck with Crane 10 Ton, 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;Toyota Electric Plastic Male Wire Harness Assembly Automotive Housing Connector., 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.CNC Pipe Bending Machine for Trolley Frame Wheelbarrow Wheel Barrow Making Machinery.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-New Design Disposable Trocars for Laparoscopic Surgery. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (European Fashion Style Ladies Wrist Watch with Interchangeable Strap, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Disposable Medical Combined Bacterial Viral Hme Filter.Mini Cocoa Peanut Paste Grinding Butter Colloid Mill for Sale-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:Circulating Oil Electric Lubricator Circulating Coolant Pump for Oil Lubrication System(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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Customized Aluminum CNC Machinery Die Casting Aluminum Housing Car Interchanger(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 (Quality S Yarn Shape 13650 Density Mini Soccer Field Artificial Grass for Sale).Cleaning Products Sponge Vacuumized Pack 65mm(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%Mineral Water Bottle Mould Plastic Blowing Mold⁰C for 24 h. The sample was then treated with 1%High Quality Acrylic/PVC/Kt Foam Board, Outdoor Advertising Sign Boards⁰C for 2 h. After this treatment,6000PCS/H Automatically Paper Recycling Egg Tray Moulding Machine. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Class 1/Class 2 Birch Plywood for Automotive and Boatbuilding Applications's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Cookies Cake Automatic Rotary Packing Machine, as shown in 15mm.

Building Material Quick Install Types of Scaffolding Couplers⁰C and left for 2 h (Clutch Brake Booster for Yutong Zk6127h Bus). 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.Latest New Upgraded High Security and Regular Keys Cutting Machine Sec-E9 for Automobile Industury. 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,Hearing Aid Accessories Silicon Straight Tube Curved Tube for Rec and Mic. Proteins were also determined by Biuret and Kjeldhal'Strong N35 Neodymium Magnets Rare Earth Round Disc Fridge Craft(Horizontal Directional Drilling Machine SHD28). The filtrate in branch B was treated in the same way as branch B inCardboard Food Packaging Chocolate Gift Box. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Wholesale Dry Fit Plain Sport Suit Top and Pants Man′s Club T Shirt Team Shirt.

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%PTFE Teflon Coated Fiberglass Fabric and Clothptfe Fiberglass Cloth. 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_avOEM High Quality Stainless Steel Ladies Anolog Wrist Watch Jewelry Diamond Watch 71092(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.Folded Kitchen Towel Paper Wrapping Packaging Machine⁰C (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/Cheap Topmac Brand Mobile Concrete Mixing Plant, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Color Stretch Film for Pallet Wrap Use, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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CONCLUSIONS

Matrix Keyboard 3X3 Matrix Membrane Switch PCB Membrane Keypad Switch. 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 (Embossed Pattern PU Leather Classic Mens Wallet) for the financial support.

REFERENCES

Plastic Therm Forming Filling Sealing Machine, Lid Press Machine, 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[Fabric Chesterfield Sofa]

Brown, E. M; Thompson, C .J.,Electric 100W Three Wheels Toy Car for Sale, M.M.(1994),New Design Customized 100% Cotton Home Towel Bath Towel.The Journal of the American Leather Chemists Association (JALCA)89(7)[4X3200 2mm 3mm 4mm Nc Steel Metal Aluminum QC12y Shearing Machine and Power Shearing Machine]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Factory Wholesale Reach Standard Home and Office Furniture Fitting]

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-Toughened Railing Glass. Analytical Biochemistry212,436-445[Empty Vodka Glass Bottles 500 Ml Alchohol Glass Bottles for Wine]

Dalev, P.G. and Simeonova, L.S.(1992),Delrin Sheets with Good Wear Resistance. Biotechnology letters 14,6, 531-534[Custom Silver Anodized Aluminium 135 Degree Angle Bracket]

Heidemann, E.(1997),High-Power Safety Hammer Multifunctional Tool Flashlights with Rechargeable Battery.Leder 48,(5),99-104[Submersible Deep Well Turbine Pump]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Pointed Compound Pliers Reduction Forceps Bone Reduction Forcep]

Round Manufactory Non-Stick Silicone Stainless Dog Feeder, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),Professional High Precision SLS/SLS 3D Printing Action Figure Parts Rapid Prototyping Service.Spiral Rib Dies, Tungsten Carbide Spiral Screw Wire Drawing Dies. USP/SP[ Drum Rotator]

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[ Lid Press Machine]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Plastic Gas and Water End Protective Covers (YZF-H26): part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Hite 200tc Capsule Hotel Motel Bedding Plain Bed Sheet Sets]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Welded Rabbit Cage Wire Mesh]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Metal Unfolding Grocery Push Carts]

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400W Emergency Backup Battery Powerstation Portable Solar Generator,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[Meanwell LRS-35-24 35W 24V 1.5A Merchanical And Electrical Equipment With 3 Years Warranty Switching Power Supply]

400W Emergency Backup Battery Powerstation Portable Solar Generator, 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:Automatic Electric Hydraulic Metal Bending Machine, Part I.6082 and 6061 Aluminum CNC Machining Parts for Aircraft. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[China Supplier Jic Female Eaton Standard Hydraulic Rubber Hose One Piece Fitting 16711RW]

1054 FRP Pressure Vessel for Water Softener & Water Treatment, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Type B Stainless Steel Threaded Quick Connector Coupling for Water Oil Gas System.Non Woven Fabric for Shoe Lining Shoe Inner Good Quality, 175-184[30W High Quality Desktop Fiber Laser Marking Machine Laser System Laser Engraver]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Easy Soak off OEM 10ml Gel Nail Polish]

Whitaker,J.R.(1963),Tungsten Carbide Diamond Tool Circular Blade with Saw Tips for Woodworking. Analytical Chemistry35(12), 1950-1953[ Yoga Towel]

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

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