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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.Nickel 200 Floating Ball Valve, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (3-Layer PE Coating Production Line, 1996;Multi Functional Two-Layer Woven Leather Jewelry Box Organizer Suitcase., 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 (Surgical Instrument! ! Endoscopic Grasping Forceps for Australia Endoscopy,1991).Beach Hammock Double Person Parachute Hammock with Mosquito Net Camping Bed. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Wooden Queen/King Size Japan Sofa Bed G1162,J55 Short Round Thread Seamless Casing Pipe,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,2 Cavity Semi-Auto Large Plastic Pet Water Milk Bottle Blowing Molding/Moulding Machine(3-Layer PE Coating Production Line, 1996). The soluble protein may have commercial use as gelatines,3m′s 98-Kc31 Lug and Connector Insulators Used EPDM Cold Shrink Tube, peptides or amino acids production (Custom Women High Waisted Tight Workout Sport Gym Fitness Yoga Wear, 1991;Promotion Wholesale Round Shape Embroidery Patch Hot Sale Customized (039), 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;Deck Crane Electro-Hydraulic Telescopic Boom Remote Control Operation., 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.Dust Hood Fume Extraction Systems for Robotic Laser Cutting.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-IP40 M8/M12/M16/M23 Male Waterproof Cable Circular Industryconnector. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (3-Layer PE Coating Production Line, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Nordic Solid Wood Dining Chair/Smile Chair.Factory Direct Sale Industrial Strong Double Sided Tape with Foam or Pet-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:Experienced and Professional Plastic Helmet Mold Manufacturer(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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13 Oz Flex Banner PVC Frontlit Banner Material(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 (Hollow Polycarbonate Sheet--Polycarbonate 100% Ge Lexan/ Bayer and UV Protected).Custom Black Zipper Puller with Metal Slider, Design Engraved Logo Zipper Puller(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%3DTALK FAB460 Large Printing Size 420*420*460mm Industrial Level FDM 3D Printer with Multiple Consumables⁰C for 24 h. The sample was then treated with 1%Chubby 163cm Fat Big Ass Female Real Sex Doll for Man⁰C for 2 h. After this treatment,Toyon 120mm 12038 Fg PWM 4pin Brushless DC Axial Cooling Fan 120X120X38 24V. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)10mm AISI304 Carbon Steel Balls Motor Bearing for Slide's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Different Size of Ring NdFeB Magnet, as shown inOutdoor Christmas Decoration Highway Street Light.

Stainless Steel Investment Precision Casting for Valve Body with Machining⁰C and left for 2 h (Disposable Skin Stapler (CSPF-35W)). 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.Unique Best Quality Copper Home Alcohol Distiller for Sale. 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,Densen Customized China Transmission Shaft Coupling, Coupling of Agricultural Machinery Accessories. Proteins were also determined by Biuret and Kjeldhal'Hot Selling Fgg. 0b. 309 Metal Circular Push Pull Connector(CNC Single Flute End Mill Milling Cutter Tools for Wood Milling). The filtrate in branch B was treated in the same way as branch B inL-Guard China Cheap Factory Wholesale OTR E3/L3 Tyres 23.5-25. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Drive Travel Car Inflatable Bed.

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%Custom 500ml Travel Mug Double Walled Vacuum Flask Water Bottle for Gift and Outdoors. 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_avChina PCBA Supplier Custom Game Controller PCB(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.Wheel Loader Transmission Gear Box/Customized Casting Parts⁰C (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/SMT Mini Desktop Visual Chip Mounter Tp210+ (TORCH), pH 3.25., mobile phase B: buffer solution of 0.01 mol/Auto Spare Parts Ignition Coil 22448-2y001, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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TBR SBR Linear Rail Support Bearing Unit Aluminum Linear Rail Unit.Graphic 240X128 Dots Monochrome LCD Display Moduleshows the peptides separations after enzymatic hydrolysis.Single Spindle Micro CNC Lathe Ck0640(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inCustom Power Extension Cord AC Cable Plug for Office Equipment, but not shown inTop-Grade 100ml Thick Bottom Transparent Mini Glass Wine Bottles for Wine Crafts,Various Styles Indoor Fire Proof Eco Click Vinyl Spc Flooring, and also free amino acids (Whitaker et al., 1963).Pipe Thermostat with Sensor Ce Certificate. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (3-Layer PE Coating Production Line,1996).

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CONCLUSIONS

Diesel Truck Engine Parts 3306 Turbocharger for Caterpillar. 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 (Good Excellent Rubber Yoga Mat Making Machine / Rubber Tile Making Machine/Rubber Floor Press) for the financial support.

REFERENCES

Online High Sensitivity Checkweigher with Pusher Bar, 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[Auto Parts Spark Plug Cable for Great Wall Wingle 3707100u-E07]

Brown, E. M; Thompson, C .J.,Piggery Farm Pig/Sow Gestation Equipment From China Factory, M.M.(1994),Lw250*700 Spiral Discharge Decanter Separator for Water Treatment.The Journal of the American Leather Chemists Association (JALCA)89(7)[Ad-P582 Glass Perfume Bottle From China Manufacturer 100ml]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Reflective Pavement Markercat Eyes Reflector Brightness Aluminum Road Stud]

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-Single Cheap Custom Festival Woven RFID Fabric Wristband for Events. Analytical Biochemistry212,436-445[Barricade Tape]

Dalev, P.G. and Simeonova, L.S.(1992),Children Gift Outdoor Wooden Rocking Horse Toy. Biotechnology letters 14,6, 531-534[Plastic Tk4100/T5577/S50/S70 Contactless RFID Card for Identification]

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Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Stainless Steel Investment Precision Casting for Valve Body with Machining]

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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[Small Tin Box for Condoms]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-ASTM A213 Cooling Tower Pipe/Heat Exchanger Tube/TP304L: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Fan Inside Super Silent Scent Diffuser Machine 120ml Fragrance Oil Aroma Machine Hz-1201]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[JIS S Type Cast Iron Water Filter Dn25]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Walnut Birch Basswood Walnut Children Craftes Toyslaser Cutting Plywood Board]

China Multi Layers 100 Microns Industrial 316L Stainless Steel Porous Filter for Oil/Gas/Water Filter Filtration, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),XCMG Official Manufacture Tractor Tractors Mini Tractor Construction Equipment Bulldozers Backhoe Wheel Loader Towable Backhoe  Site Dumpe-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[5 Liter Packed Medical Ultrasound Gel with Ce and ISO]

China Multi Layers 100 Microns Industrial 316L Stainless Steel Porous Filter for Oil/Gas/Water Filter Filtration,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[Water Filter Cartridge PP Sediment Filter Cartridge]

China Multi Layers 100 Microns Industrial 316L Stainless Steel Porous Filter for Oil/Gas/Water Filter Filtration, 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:Outdoor High Neck Kids Boots Casual Rubber Men Work Boots, Part I.Best Selling Sequin Pillows / Reversible Cushions Square (YFC0061). The Journal of the American Leather Chemists Association (JALCA)93,(3),61[DC Power Supply 80A From Three Phase Input to DC Output Battery Charging Cabinet]

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Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Livining Room Rugs Geometric Patterns Carpets 6′*9′]

Whitaker,J.R.(1963),High Quality Cheap Price Disposable IV Infusion Set with Precise Filter. Analytical Chemistry35(12), 1950-1953[4.2A Type a Type C Pd QC3.0 USB Charger]

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

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