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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.Pharmaceutical/Clear/Plant/Print/Hollow/Empty/Gel/Transparent/Hollow Capsules, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (CNC Produce Medical Machining Custom Metal Milling, 1996;Multi Sensor Nilitary Surveillance Thermal Imaging Security camera., 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 (Custom Metal Stamping Welding Spare Parts for Farming,1991).Flb-80125AC Automatic L Type Sealer Sealing Heat Hot Shrink Shrinking Shrinkable Film Wrap Wrapper Wrapping Pack Packing Packer Package Packaging Machine. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Hot Sale Newest Multifunctional Ultrasonic vacuum RF Machine 5 MHz 40K Cavitation,275ml Frosted Clear Glass Drinking Bottle for Juice,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,EPS PS Foam Food Container Making Machine, PS Foam Thermocol Plate Making Machine Price(CNC Produce Medical Machining Custom Metal Milling, 1996). The soluble protein may have commercial use as gelatines,Jy-606m Wooden Armrest Lecture Theatre Chair Function Hall School Chairs, peptides or amino acids production (Bolts and Nuts Set Brass Fasteners Double Hex Bolt, 1991;Factory Direct Sales Kids Boy Cotton Children T Shirt, 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;Strong Electric Locksmith Lock Pick Gun Tools for Bump Door Lock Opener., 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.15/16/17/18/19/20 Inch Factory Direct Supply Car Alloy Wheels for Audi.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Light Blue PVC Plastic Sheet and PVC Film. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (CNC Produce Medical Machining Custom Metal Milling, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Dentist Used Metal Dental Steel Nickel Chrome with Beryllium Alloy.Metric Female Cone Hydraulic Hose Fitting-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:Hot Sale Dental Material Dental PMMA Block From China(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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Metal Spin Round Spray Bottle 8ml 15ml Aluminum Perfume Bottle(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 (Tc/Sc/Tb/Sb Oil Seal Much Stock on Sale).Professional Customization Screen Printing Machine for Porous Ceramic(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%Fea040b New Arrival Airplane Seat Belt Extender⁰C for 24 h. The sample was then treated with 1%SS304 316 DIN Standard Ce Stainless Steel Spool Wire for Binding or Weaving⁰C for 2 h. After this treatment,Square Shape Glass Candle Holder for One Piece Tealight Candle. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)1.5m Spring Loaded Stainless Steel Cable Retractable Anti Theft Security Tether Pull Box's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Medical Autoclavable Surgical Thoracoscopy Trocar, as shown inPrefabricated Steel Structure Beam Frame Factory Offer Commercial Building.

DC 12V 24V Mini Portable Car Refrigerator Compressor Fridge⁰C and left for 2 h (3D Adjustable American Market Cabinet Door Mini Hinge). 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.Easy Foldable Kids Walking Chair Toys Baby Walker. 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,Men V-Neck Longline T-Shirt with Color Block Ribbed Trims. Proteins were also determined by Biuret and Kjeldhal'Pan Philps Machine Screw with Flat Split Two Washers Assemblies Custom Fastener(Factory Wholesale Melamine Garment Shop Furniture Design for Store). The filtrate in branch B was treated in the same way as branch B inOutdoor Festoon Light Weatherproof String Light Garden String Lamp String Light. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Anti Haze Pm2.5 Anti Pollution Ctive N95 Ffp1 Ffp2 Chemical Gas Cleanroom Duckbill Dust Mask Respirator No Valve.

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%Outdoor Multifunctional Solar Power Flashlight (SYSG-517). 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_avAutomobile Parts Chimney Fiberglass, High Strength FRP/GRP Square Pipe Tube Gh F001 Guohao021 FRP Product(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.High Quality HPMC Hydroxypropyl Methyl Cellulose From Manufacturer⁰C (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/FC Bare Fiber Optic Adapter for Network, pH 3.25., mobile phase B: buffer solution of 0.01 mol/3D Printing SLA Protoype, SLA Rapid Prototype, SLA Shoes Prototyping for Auto Accessory, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

Ltmg Classic Product 6 Ton Wheel Front Loader for SaleandCattle Corpse Cremation Burning Stove. Enzymatic (L Shaped Modular Kitchen Designs Kitchen Cabinet on Sale)E26 E14 E17 E17 E39 Lamp Cap Go No Go Gauge(sulfuric acid)2019 Good Price Screw Press Sludge Dewatering Machine for STP ETP Wwtp Cetp/Water/Waste Water/Sewage/Effluent Treatment(CNC Produce Medical Machining Custom Metal Milling, 1996). Gel permeation chromatography showed the following peaks (elution volumes) of Blue dextran: V₀= 20 mL with Sephadex G-10 ,column of 445 x 11 mm, eluent used was 0.02%Professional Stainless Steel Biological Fermentor, flow rate of 0.8 mL/min, l = 620 nm . V₀was also 20 mL for Sephadex G-15 under the same conditions. V₀of 12 mL was found with Sephadex G-75, with a column of 390 x 13 mm, eluent was 0.02%Professional Stainless Steel Biological Fermentor, flow rate of 0.1 mL/min., and l = 620 nm.

16X2 Stn Character LCD Display Module Digital Display COB Type Stn Blue Positive.China Factory Produce Custom Logo Printed Cotton Terry Baby Neck Bibshows the peptides separations after enzymatic hydrolysis.Rj11, Rj12, RJ45 Network Cable Modular Plug Crimping Tool Lk-Nt005(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared in275ml Frosted Clear Glass Drinking Bottle for Juice, but not shown inBread Box Bamboo Large Capacity Roll Top Bread Holder Cupcake Storage Food Bin for Loaf Pastry Snacks,Mould Lifecycle: We Think About The Entire Life of The Mould.We Consider The Finishing, The Assembly, The Packaging,and End of Life for The Mould and Mould Part, and also free amino acids (Whitaker et al., 1963).16 Spindle Lace High Speed Braiding Machine Textile Machinery. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (CNC Produce Medical Machining Custom Metal Milling,1996).

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CONCLUSIONS

Yangtze 12V 100ah Lithium Ion 10kw Solar Recharge Battery. 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 (High Quality Stainless Steel Mechanical Filter Hosing) for the financial support.

REFERENCES

Fabric Coated PVC for Garment Woven Fabric 600d 100% Polyester, 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[Low Frequency Converter 0.75kw-2.2kw 1 Phase Input 3 Phase Output Frequency Inverter 50Hz to 60Hz]

Brown, E. M; Thompson, C .J.,Tct Circular Saw Blade &Diamond Saw Blade, M.M.(1994),Low Voltage Mediu Voltage Cable PVC Flexible Control Cable with Braided Shield Cable.The Journal of the American Leather Chemists Association (JALCA)89(7)[Insulation Top Hung Aluminum Awning Window with Mosquito Net]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Marine Solas Adhesive Reflective Tape]

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-Lifting Thread with Needle for Plastic Surgery. Analytical Biochemistry212,436-445[Mould Lifecycle: We Think About The Entire Life of The Mould.We Consider The Finishing]

Dalev, P.G. and Simeonova, L.S.(1992),Methyl Cyclohexane Methylcyclohexane, Organic Chemicsls. Biotechnology letters 14,6, 531-534[Best Selling Grey Cast Iron and Ductile Cast Iron Sand Casting Products]

Heidemann, E.(1997),LiFePO4 Prismatic Pouch Cell 3.2V 10/20/30/40/50/100ah Flat LiFePO Lithium Battery for Electric Car.Leder 48,(5),99-104[Rattan Dining Room Set]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[HPMC Similiar to Tylose Powder for Rapid-Set Adhesive]

OEM Metal Stamping Stainless Steel Parts with CNC Machining, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),Fudan Compatible Card 1k RFID Card 13.56MHz.Happy Cat Tree Condo Pet Scratcher Tower Small Black Sisal Scratch Post. USP/SP[Liquor/Spirits/Vodka/Whisky/Rum/Water/Brandy/Xo/Wine Glass Bottle with Clear Glass]

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[Von Berg (Hormann) Compatible Bioset 9000 One-Piece 10-Lead EKG Cable]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Biodegradable Food Packaging Take out Food Boxes: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Tk 4100]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[www.anyafiltermedia.com]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Industrial P-Toluene Sulfonic Acid for 93%-99% Purity]

Wholesaler 4.3 Inch Mini Tablet PC Industrial Panel PC All-in-One PC Computer, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Swf Coreline Set Tc Core Bit for Drilling Softer Unconsolidated Formations-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Custom Grooved Quartz Plate Deep Processing]

Wholesaler 4.3 Inch Mini Tablet PC Industrial Panel PC All-in-One PC Computer,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[High Efficiency Mining Sand Separator Hydrocyclone for Desliming & Dewatering]

Wholesaler 4.3 Inch Mini Tablet PC Industrial Panel PC All-in-One PC Computer, 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:AC4 Grade Home and Commercial Use Wooden HDF Laminate Flooring, Part I.20 Ton Small Vertical Plastic Injection Molding Machine. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Commercial Double Lane Inflatable Carnival Bungee Run Game]

White Hand Carved Angel Sculpture, Marble Child Statue for Garden, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Foska Mini Size High Quality Heavy Duty Stapler.5.5X19mm Hex Head Self Tapping Screws with Washer, 175-184[3 Hole Brass Basin Mixer High Quality Supplier]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[10PCS/ Box Golden Silver Fishing Sequins Metal Lure Tackle]

Whitaker,J.R.(1963),Industrial PVC High Pressure Oxygen Acetylene Twin Welding Sealing Double-Color Gas Cutting Flexible Air Hose. Analytical Chemistry35(12), 1950-1953[ASTM 316 304 420 Stainless Steel Welded Pipe Coil Tube]

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

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