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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.Auto Iridium Spark Plug for Toyota 90919-01253 Sc20hr11, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Automatic Soap Bag in Box Packing Machine, 1996;ETL cETL Dlc 150 Watt LED Street Light for Parking Lot Lighting., 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 (High Performance Alcohol Pad Making Machine,1991).Ss304 Sanitary Welded Butterfly Valve. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,2019 New Design Flat Panel Print 3D Microfiber Cartoon Bedding Set,Anti-Abrasion PVC Artificial Leather for Car Seat (128#),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,Digital Control with LED Display DC Solar Small Deep Freezer Fridge(Automatic Soap Bag in Box Packing Machine, 1996). The soluble protein may have commercial use as gelatines,Automatic Economical Check Weigher with High Speed, peptides or amino acids production (Small Lovely Girls′ Backpack Plush Backpacks with Stitching Pocket, 1991;Steel Handle Rake Flat Rake Garden Rake Agricultural Rake, 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;High Quality Small Tea Bag Packing Machine Price., 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.Carbon Dioxide Gas with 50L Seamless Cylinder.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-UV8d Wouxun High Gain Antenna Dual Band Antenna. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Automatic Soap Bag in Box Packing Machine, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Fiberglass Covered Insulated Flat/Square Aluminum Magnet Wire.Portable EMS Shock Wave Massage and Sport Pain Therapy Equipment Beauty Machine-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:Swbf 7b2s Bolt Former, Bolt Machine, Cold Forging Machine, Cold Heading Machine, Parts Former, Nut Tapping Machine, Threadrolling Machine, Automatic Bolt Making(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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Ce TUV SGS BV Certificate CNC Router with Rotary Fixture China(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 (Stainless Steel Sanitary SS304/SS316 2 Way Tri Clamp Ball Valve).Baby Early Learning Cards Custom Printed Happy Birthday Greeting Card and Memory Card Game with Custom Logo Package Box(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%Optical Fiber Connector Cleaning Cassette Reel Cleaner, Non-Alcoholic Specialty Fibers 500+ Cleans⁰C for 24 h. The sample was then treated with 1%Characteristic Impedance of 75 Ohms High-Foaming Coaxial Cable Rg59, RG6, Rg11 for Home and Commercial Application, Audio and Video Wire⁰C for 2 h. After this treatment,Modern Container House / Modular Residential / Prefabricated Mobile Home. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)100% Polyester Luxury Bedding Comforter Fleece Blanket Throw's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Aluminum Window System Thermal Break Strip (HK Stype35.3mm), as shown inDurable Inline Hockey Skate Puck.

Multi-Function Knife CREE XPE 5mode Rechargeable LED Flashlight⁰C and left for 2 h (China High Profit Automatic Pet Extruder Machine). 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.Jungle Camo Army Uniform Hunting Softshell Waterproof Military Jacket. 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,Full Automatic Milk/ Chemistry/Shampoo Plastic Bottle Blowing Machine. Proteins were also determined by Biuret and Kjeldhal'High Speed Mixer for Plastic Extrusion Line(Automatic Economical Check Weigher with High Speed). The filtrate in branch B was treated in the same way as branch B inGuide Rail Cargo Lift. The final soluble protein shown were used for peptides separations using gel permeation chromatography,High Quality Trailer Wire Harness with 7 Pole Trailer Plug & Fuse Holder for Trailer Wiring Harness Assembly for Challonger.

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%10mm Gold Plated Stainless Steel Balls G100 Polish Ball. 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_avCold Rolled Bearing Ball Screw Sfu4005 for CNC Machine(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 - 5.5.304 Seamless Stainless Steel Tube⁰C (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/Stainless Steel Manual 3-Way Plug Valve, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Hanger Rail Shot Blast Cleaning Equipment/Suspended Shot Blasting Machine, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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CONCLUSIONS

450mm High Quality Circular Diamond Cutting Saw Blade for Marble Stone. 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 (P10.4 Outdoor LED Transparent Display Screen Glass Curtain Screen) for the financial support.

REFERENCES

Manual Glass Lifting Machine for Making Double Glass, 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[Genuine Leather Handbag Women Shoulder Bag Lady Crossbody Bag]

Brown, E. M; Thompson, C .J.,Aluminium Profile Heavy Duty Bending Machine Aluminum Window Door Machine, M.M.(1994),Towed Bitumen Mixer Slj5.The Journal of the American Leather Chemists Association (JALCA)89(7)[Medical Supplier Non Woven 3ply Earloop Disposable Face Mask]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Lab Water Instruments/Lab Water Purification System Micro]

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-High Adsorption Capacity and Porosity Chloride Adsorbent Activated Alumina Price. Analytical Biochemistry212,436-445[Proper Price Good Quality 10ml 20ml Airless Pump Bottle]

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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[Atlas Copco Compressor Spare Parts Service Kit 2901074800 2904015600 2906009600]

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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[Dimmable Industrial Indoor Commercial Indoor LED Highbay Lighting for Warehouse]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Viscose Sodium Sulphate Anhydrous by Sateri: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[LED Cabinet Light Wardrobe Furniture Sink Drawer Lighting Work Lamp]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[American Bathroom Vanity]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Winter Design Acrylic Beanie /Bobble Jacquard Hat with Flowers /Machine for Knitted Winter Cap with Embroidery Circular Hat Knitting Machine]

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Orange Silica Gel 2-4 mm with High Quality,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[Light Weight Ohsas 18001 Certified Metal Arch Buildings]

Orange Silica Gel 2-4 mm with High Quality, 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:aluminium discs for fry pan alloy 1060 1100 1050 3003, Part I.High Quality Insulated Double Wall Bamboo Glass Water Bottle. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[High Performance Alcohol Pad Making Machine]

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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[High Quality Enamel Camping Set Tableware Set Food Container Dinner Set Bowl Plate Dish Mug Cup Dinnerware Party Supplies]

Whitaker,J.R.(1963),I10 Tws Mini Bluetooth 5.0 Earphone Twins Wireless Headphones Air Pods in Ear Earbuds Sport Headset Stereo for Apple iPhone Samsung Xiaomi. Analytical Chemistry35(12), 1950-1953[Reusable CAT6 UTP RJ45 8P8C Modular Plug]

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

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