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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.Car Interior Accessories Customized Car Seat Cover, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Custom PVC Plastic Card Printed School/Student/Office/Staff/PVC/ID/Name/Business Cards with Your Design, 1996;10 X 10 Canopy Pop up Tent Outdoor Gazebo Tent for Exhibition., 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 (Clean Room Purifying Equipment NSF Certified Biological Safety Cabinet,1991).Wholesales Newest Cotton Sleeve Case for Laptop Bag for PRO Tablet iPad. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,OEM/ODM Raw Material MDF HDF Laminate Door,7628 Fiberglass Cloth/Fabric,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,Stainles Steel Cartridge Filter(Custom PVC Plastic Card Printed School/Student/Office/Staff/PVC/ID/Name/Business Cards with Your Design, 1996). The soluble protein may have commercial use as gelatines,750ml Long Neck Dark Green Color Glass Bottle for Wine, peptides or amino acids production (Floral Kids PVC School Raincoat, 1991;MB-Z700 10-12t/H Hot Sale Wood Log Debarking Machine, 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;Fs2121 Locking Latch Flush Stainless Steel and Flush Paddle Latch with Truck Toolbox Lock., 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.Microsynthetic Fiber Virgin Homopolymer Polypropylene Monofilament Fibermesh.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-78mm Aluminum Gold Color Plastic Jar Screw Cap. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Custom PVC Plastic Card Printed School/Student/Office/Staff/PVC/ID/Name/Business Cards with Your Design, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Best Price 100kVA/150kVA Cummins Power Generator Diesel Engines.1008 Spindles Auto Doffer Machine for Spinning Ring Frame Polyester Yarn Winding Spinning Machine Cotton Wool Thread Ring-Spun Yarns Making 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:0.5L Plastic Pet Water Bottle Blower Molding Machine/Pet Bottle 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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Concrete Electric Pole Pile Making Spinning Machine(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 (Extendable 485mm Stun Guns Baton with Electric Shock for Self Defense (X10)).Shop Front Indoor Durable Hanging Banner Advertising Signs(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%Cosmetic & Food Grade Sodium Hyaluronate⁰C for 24 h. The sample was then treated with 1%High Frequency Circular Vibrating Screen⁰C for 2 h. After this treatment,Fiberglass/Composite Mesh for Walkway FRP Grating. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)High Quality 33kv 6.3kv 1000 kVA 50% Copper and 50% Aluminum Material Distribution Transformer Made in China's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Us FDA Authorised 1g Desiccant Silica Gel Used for Food/Medicine, as shown inGlow in Dark Ceramic Sublimation Mug 11oz.

Union NPT Bsp BSPT Stainless Steel Pipe Fittings⁰C and left for 2 h (printing Rehabilitation of protective). 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.High Brightness IP67 Red Color SMD5050 Chip 60leds 14.4W DC12V LED Strip. 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,Marine Anemometer / Wind Meter/ Anemoscope with Alarm Function for Vessel/Ship. Proteins were also determined by Biuret and Kjeldhal'2019 Hot Sale High Quality Chinese Factory Direct Supplied Heavy Duty 3-200ton Electric Winch(Multi-Function Knife CREE XPE 5mode Rechargeable LED Flashlight). The filtrate in branch B was treated in the same way as branch B inHigh Quality CNC Machined Motorcycle Brake Caliper with Any Colors. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Tear Strip Tape.

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%Healthy Grooming Kit for Caring Newborn Baby. 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_avReusable Stainless Steel Metal Collapsible Portable Foldable Telescopic Straw with Silicone Tips Cleaning Brush(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.Private Label Fitness Wear Sublimation Printing Cheap Long Womens High Waist Yoga Pants Custom⁰C (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/Emergency Stop Switch 880 Lbs Lift Electric Hoist Crane Remote Control Power System, pH 3.25., mobile phase B: buffer solution of 0.01 mol/Auto Valve 4-20mA Flow Control Butterfly Valve, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

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Stainless Steel LED Underground Light, Ground Lighting IP66.Fashion Children Fur Pompom Beanie Scarf Knit Setshows the peptides separations after enzymatic hydrolysis.GMP Certified Health Food Vitamin C Electrolyte Effervescent Tablets (4G/ tablet * 10 tablets)(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared inK-Pop Bts Baseball Hat Sport Anime Cap, but not shown inPet Material Alien 9654 Wet Inlay UHF Windshield Label,K Series Helical Bevel Reduction Gearbox Motor, and also free amino acids (Whitaker et al., 1963).OEM Service Ce/ISO Approved Endoscopic Disposable Grasping Forceps. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (Custom PVC Plastic Card Printed School/Student/Office/Staff/PVC/ID/Name/Business Cards with Your Design,1996).

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CONCLUSIONS

Luxurious Fire and Burglary Proof Hidden Hotel Security Safe Box. 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 (Ce Approved Bowrider Leisure Boat for 6-8 People) for the financial support.

REFERENCES

Fst800-211 Industrial Water Pressure Sensor and Transmitter with 4-20mA Output, 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 Cheap Chinese Healthy Watermelon Kernels Food Ingredients]

Brown, E. M; Thompson, C .J.,14 Head Multihead Weigher for Pork with Screw Feeding, M.M.(1994),Stainless Steel Sanitary SS304/SS316 2 Way Tri Clamp Ball Valve.The Journal of the American Leather Chemists Association (JALCA)89(7)[Plastic Nylon Sheet with High Temperature]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Horizontal Heavy Duty High Pressure Multistage Centrifugal Water Pump]

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-PSF 1000 Flat Plate Upper Discharge separator. Analytical Biochemistry212,436-445[High Lumen Outdoor IP65 100W LED Flood Light]

Dalev, P.G. and Simeonova, L.S.(1992),Aromatherapy Essential Oil Ultrasonic Diffuser with APP Remote Control Smart. Biotechnology letters 14,6, 531-534[Fiber Optic Equipment Sc/APC Quick Connector Sc Fast Connector Field Assembly Connector]

Heidemann, E.(1997),Powerful Neodymium Pot Magnet Rare Earth Pot Magnet.Leder 48,(5),99-104[19 Inch Best Service Good Quality Competitive Price Rack Cabinet]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Rapid Prototype Plastic Molding Simple Rapid Tooling and Simple Mould]

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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[1220*2440*15mm Chipboard with Plain Face for Furniture Decoration From China]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-High Whiteness & Excellent Dispersity Anatase Type Titanium Dioxide: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Antique Stainless Steel Sheets Thailand 430 Stainless Steel Sheet]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Coconut Charcoal Toothpaste Kits]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[99% Al2O3 High Alumina Ceramic Ball]

Large Jewelry Display Box for Ring Necklace Earring Storage Case, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Indoor Tub Customized Shower/Free Standing Shower Enclosure-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[Deluxe Saddle Doctor Chair Dental Stool with Adjustable Seat]

Large Jewelry Display Box for Ring Necklace Earring Storage Case,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[M16 M20 Stainless Steel Hilti Chemical Thread Bar Anchor Bolt Wedge Hex Bolt Sleeve Anchor Grade 5.6]

Large Jewelry Display Box for Ring Necklace Earring Storage Case, 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:Glow in Dark Ceramic Sublimation Mug 11oz, Part I.Double Poles Number Bimetal Strip 1AMP Mini Circuit Breaker. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[0.5L Plastic Pet Water Bottle Blower Molding Machine/Pet Bottle Making]

Stainless Steel 201/304/316 CF8/CF8m 1000wog Threaded Ball Valve, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),22-30L/H Commercial 3 Flavors Mix Soft Ice Cream Cones Maker Machine.Supermarket Combined Island Chest Freezer Commercial Refrigerator, 175-184[4axis Wood Router]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Full Automatic Combination Weighing Packing Machine Jy-420A]

Whitaker,J.R.(1963),PLA Biodegradable Compostable Plastic Lids Eco-Friendly with Factory Price. Analytical Chemistry35(12), 1950-1953[Foshan Terracotta Facade Panel for Commercial Building Curtain Wall]

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

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