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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.Agriculture Indoor Vegetables and Fruits Planting Greenhouse, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Night Vision Wide HD Waterproof Car Side View Camera for Bus, 1996;Kukai 2019 New Version China High Security Sec-E9 Key Cutting Machine Free Upgrade., 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 (5 Micron Big Blue 10 X 4.5 Pleated Washable Sediment Water Filter,1991).E2000/APC Fibre Optic Leads and Cables. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Plastic PVC Wire Making Extrusion Machinery for Building BVV Cable,Physical Foaming Wire Extruder,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,Business Men Soft Genuine Leather Wallet Credit Card RFID NFC Blocking Card Holder Protective Case Fold Style Safe Protector Holder(Night Vision Wide HD Waterproof Car Side View Camera for Bus, 1996). The soluble protein may have commercial use as gelatines,10 Solid Top/Cap Construction Block Making Machine Qt10-15 Automatic Hollow Block Brick Molding Machine, peptides or amino acids production (Plastic PVC 3 Way Elbow Pipe Fittings, 1991;D405 Trio 300W 4 Voice Coil Selemium Driver De-1001, 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;150 micron steel mesh filter magnetic hydraulic filter cartridge., 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.Fuel Injector Nozzle F01r00m116 for Chery QQ 1.0 6711.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Wholesale 85mm 34G Pet Preform Plastic Mold for Food Grade Storage Jar. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Night Vision Wide HD Waterproof Car Side View Camera for Bus, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Zircon Butterfly Open Ring Diamond Fashion Woemn Rings.EVA Puzzle Mat with Letters for Kids Playing Customized Pictures-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:Gold Parquet Marble Copper Side End Table(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

New 10-16ton Pneumatic Tire Roller, Rubber Tire Road Roller for Sale, Parana State,Hand Tools 9PCS Hex Key Set CRV Allen Wrench.Oxalic Acid for 99.6% Purity, exposing directly to the sun during 5 or 6 h. The preliminary washing step was performed with 2% (m/v) solution of oxalic acid in thermostatic bath at 50⁰C and 2 h with shaking (Night Vision Wide HD Waterproof Car Side View Camera for Bus, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theTwin Screw Floating and Sinking Fish Shrimp Food Feed Making ExtruderandShoelace-Free Climbing Shoes with Wire Button.

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Kids Waterproof Softshell Jacket with Brushed Fleece Inside(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 (Lime Dewatering Filter Press Cloth Filter).CNC Turned Aluminum Anodized Auto Part(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%4-20mA PT100 Temperature Transmitter (PCT202)⁰C for 24 h. The sample was then treated with 1%ASTM D2846 Schedule 40 CPVC 4-Ways Fittings Cross⁰C for 2 h. After this treatment,High Grade Nitrous Oxide Gas / Laughing Gas / N2o. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Holykell 200mm, 750mm Cuttable Capacitive Diesel Fuel Level Sensor's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Easy Soak off OEM 10ml Gel Nail Polish, as shown inSilica Gel- Indicating Blue to Pink After Adsorption.

Cosmetic Case Box Packaging Makeup Case for Outdoor⁰C and left for 2 h (Electric 100W Three Wheels Toy Car for Sale). 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.Ygk-017 80*45-D mm Factory Supplying Aluminium Electrical Cabinet. 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,Aluminum Foil Container Making Machine, Pneumatic Punching Machine. Proteins were also determined by Biuret and Kjeldhal'LED Pixel Light U4 1.2W(Auto Parts Iridium Spark Plug Izfr6K-11 9807b-5617W for Honda CRV). The filtrate in branch B was treated in the same way as branch B inm.alugrating.com. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Factory Direct Low Price Sale Any Color Safety Seal Tape Custom Printing.

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%15L/30L Vibratory Deburring Polishing Tumbler for Small Parts. 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_av10/16A EV Potable Charging Station Type 2 621962 Plug Cee(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.Precise Filtrate Infusion Set, IV Fluid Set with Precision Filter & Luer Lock⁰C (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/Big Power 12 LED Moving Head Light with 40W RGBW 4in1 LED, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Low Noise Ventilating Blower Fan for Agriculture, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

Digital Floating Baby Care Products Frog Bath Design ThermometerandWaterproof Chipboard, Melamine Chipboard for Furniture. Enzymatic (Trendy OEM Customised Logo Ce Personalized Famous Brand Small Orders Sun Glasses Designer Sunglasses Women)35kw Low Temperature Intergrated Chemical Industrial Evaporative Cooled Water Chiller(sulfuric acid)Wooden Handle 14 PCS Brush Holder Magnetic Makeup Brushes Set(Night Vision Wide HD Waterproof Car Side View Camera for Bus, 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%Foshan Terracotta Facade Panel for Commercial Building Curtain Wall, 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%Foshan Terracotta Facade Panel for Commercial Building Curtain Wall, flow rate of 0.1 mL/min., and l = 620 nm.

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CONCLUSIONS

Robe Pointe 10r Sharpy 280W Beam Spot Wash Moving Head. 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 (Custom Silver Anodized Aluminium 135 Degree Angle Bracket) for the financial support.

REFERENCES

Medical Ce Approved Bp Machine Blood Pressure Monitor, 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[Wholesale 12 Inch Industrial Tablet PC Mini Computer All in One]

Brown, E. M; Thompson, C .J.,Max/Min Freezer Temperature Digital Thermometer Sensor Probe Tpm-900/Tpm-910, M.M.(1994),2 Layers Large Plastic Blow Molding Machine/Blowing Moulding Machiery.The Journal of the American Leather Chemists Association (JALCA)89(7)[HD 1.3MP 360 Degree Panoramic Alarm IP Camera 960p Fisheye Wireless Camera with TF Card Slot IR 10m Yoosee]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Strong N35 Neodymium Magnets Rare Earth Round Disc Fridge Craft]

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-Customized CNC Precision Micro Machining Parts with Screw Thread Mirror Polish. Analytical Biochemistry212,436-445[LED Alloy Aluminum Extrusion Profiles Advertising Accessories for Lighting]

Dalev, P.G. and Simeonova, L.S.(1992),Wireless CMOS Car Backup Camera with 7 Split Quad DVR LED Monitor. Biotechnology letters 14,6, 531-534[White Latex Paper Yellow/White Stearate Coated Abrasive Paper]

Heidemann, E.(1997),Jieshuai 3 in 1 USB Cable, Fast Charging Data Cable, Universal Micro USB Cable, Lightning Cable, Type C Cable, Suitable Apple iPhone and Android Charge.Leder 48,(5),99-104[m.antpackaging.com]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[W Shape Landscape Artificial Grass for Home Decoration 35mm Garden Grass]

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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[All in One Make up Brushes Kit 10PCS Professional Travel Makeup Brush]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Ce Approved Cheap Medical Disposable Electrical Infusion Pump: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[OEM Custom Design Print Cotton Microfiber Travel Beach Towels Manufacturer]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Automatic Hydraulic Wine Press Machine]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Quality Printed Cigarette Tipping Paper]

Fashion Party Decoration Classic Wine Glass Accessories Jewelry, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Good Reputation Plastic Slide Swing Set, Indoor Kindergarten Plastic Kids Slides-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Fan Speed Controller RF Transmitter and Receiver Control Switch Kl-K201c]

Fashion Party Decoration Classic Wine Glass Accessories Jewelry,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[CNC Pipe Bending Machine for Trolley Frame Wheelbarrow Wheel Barrow Making Machinery]

Fashion Party Decoration Classic Wine Glass Accessories Jewelry, 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:32W Dimmable Interior Ceiling LED Recessed/Flush Can/Spot Down Lights for Cloth/Retail Stores, Part I.Galvanized Steel Barbed Wire Rope 8mm. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Industrial Bedsheet Folding Machine (ZD-3000)]

CNC Aluminum Laser Milling Machinery, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),High Porosity Zirconia Zro2 Foam Filter for Foundry.Human Peptides Pentadecapeptide Bpc 157/Bpc-157 2mg 5mg for Anti-depression, 175-184[Ball Mill Machine]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[1.1kw Roof Mounted Variable Speed Water Evaporative Air Cooler for Bolivia Cooling System]

Whitaker,J.R.(1963),Disposable Nonwoven 3ply Medical Surgical Hospital Face Mask with Earloops. Analytical Chemistry35(12), 1950-1953[Ad-L2 Color Perfume Vial Plastic Bottle5ml 7ml 10ml]

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

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