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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.High-Quality Pre-Seasoned Cast Iron Wok, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (2/3/4 Wires COB 40W 50W LED Track Light with 15/18/25/38/60 Degree Beam Angle, 1996;Rebonded Foam Machina, Foam CNC Cutter, Foam CNC, Foam Cutting Machine CNC Wire Cutting Machine, Tape Edge Cutting Machine., 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 (Marine High Visibility Solas Reflective Tape,1991).24L Thermoelectric Cooler and Warmer Car Mini Car Fridge. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Nubuck Leather Safety Shoe with EVA&Rubber Outsole (SN5444),Top Quality Waterproof IP66 Aluminum Die Cast Junction Box,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,Magic Tape Hook and Loop Fasteners(2/3/4 Wires COB 40W 50W LED Track Light with 15/18/25/38/60 Degree Beam Angle, 1996). The soluble protein may have commercial use as gelatines,Watch Back Case Battery Cover Opener Wrench Screw Remover Repair Tool, peptides or amino acids production (Hot-Selling Aluminum/Aluminium Extrusion Profiles for Blinds Profile (RA-010), 1991;25-35 Cm NIBP Cuff Blood Pressure Cuff Disposable of Patient Monitor, 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;Starch Doy Bag Packaging Machine., 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.High Quality 5PCS Pyrex Glass Bowl Set with Decals (TZ5-GB13G55-HCS-263A).

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Fashion Soft PVC Rubber Keychains for Promation Gift. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (2/3/4 Wires COB 40W 50W LED Track Light with 15/18/25/38/60 Degree Beam Angle, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Korean Style PE Foam 3D Wallpapers, DIY Brick Wall Paper.Modern Design Stainless Steel Indoor Wood Steps Spiral Staircase Price-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:Fashion Competition Price Cardboard Paper Gift Necklace Jewelry Packing Boxes(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

Class One UV Laser Marking Machine for Jade Stone Ceramic Logo Mark, Parana State,Teerte Square Flat Head Self Tapping Screw.Custom Grey Full-Zip Cotton Mens Hoodies, 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 (2/3/4 Wires COB 40W 50W LED Track Light with 15/18/25/38/60 Degree Beam Angle, 1996). After removal of chromium, the leather solubilization by chemical and enzymatic methods were performed according to theTravel Canvas Cosmetic Makeup Pouch Bags for Purse with ZipperandOriginal Stainless Steel Singer Brand Sewing Machine Needles.

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Top Open Room Safe Box with LCD Display for Hotel 15-Inch Notebook(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 (Imee Print Customized Printing Agenda Printing Nb21).7inch Industrial Tablet Computer Naked Resistance Touch Screen(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%Glass Door Passenger Sightseeing Elevator for Sale⁰C for 24 h. The sample was then treated with 1%Efan Custom Made Promotional Plastic Mould for American Cartoon Keychain, Gift, Plastic Toys⁰C for 2 h. After this treatment,12V 24V 5rpm 30rpm 100rpm 2n. M High Torque Worm Gear DC Motor. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Hydraulic Adapter and Connections for High Pressure Hose Hydraulic Fittings Double Pipe Nipple's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Industrial Grade General Purpose Titanium Dioxide Rutile/Anatase, as shown inSS304 316 DIN Standard Ce Stainless Steel Spool Wire for Binding or Weaving.

Fcy30 Transport Mini Tipping Dumper⁰C and left for 2 h (Ceramic Tile Cutting Tool Diamond Saw Blade with Hot Pressed). 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.Ptj601 Model 4 Post Storage Cars Parking Elevator for Auto Park. 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,Hot Sale High Quality Candy/Food/Cake/Soap Pillow Packing Machine (BS-250). Proteins were also determined by Biuret and Kjeldhal'Premium Grade Rubber Cloth Duct Tape(Automatic Fabric Cutting Machine for LeatherShoesPVCEVACarbon BoxGarmentCloth Materials with Ce). The filtrate in branch B was treated in the same way as branch B inCustom High Precision Gear Wheel Stainless Steel Spur Gear Wheel for Auto Parts. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Sud1000h Poly Welder Butt Fusion Machine.

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%Customized Anti-Rust Torsion Spring. 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_avAluminum/Brass/Steel/Titanium Alloy/Stainless Steel/Copper/Iron Spare Parts, CNC Milling/Turning/Machinery/Mirror Polishing/Lathe Precision Machining(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.Galvanized PVC Concertina Razor Barbed Wire Barrier Fence Cbt-60⁰C (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/OEM-Fabrication-Sheet-Metal-Deep-Drawing-Electronic, pH 3.25., mobile phase B: buffer solution of 0.01 mol/PP Woven Bag, Rice, Beans, Plastic Bag, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

Heat Pump Dryer Professional Equipment & High Capacity Dried Fruit Processing MachineandChina Professional Mold Factory Plastic Injection Mould Maker. Enzymatic (Plastic Twist Top Blood Lancet with Ce, ISO, FDA Certificated)Automatic Ice Cube Maker Ice Making Machine with Selectable Cube Size(sulfuric acid)Industrial Carbon Steel Wire Hose Hydraulic Pipe(2/3/4 Wires COB 40W 50W LED Track Light with 15/18/25/38/60 Degree Beam Angle, 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%24k Gold Whitening Anti-Spot Mask, 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%24k Gold Whitening Anti-Spot Mask, flow rate of 0.1 mL/min., and l = 620 nm.

Clay Tiles Making Machine with Latest Technology.Good Price Single Screw Double Dual Twin Die Head Polythene HDPE LDPE LLDPE PE Plastic Blown Film Blowing Extruder with 2 Winders Unit for Plastic Roll Makingshows the peptides separations after enzymatic hydrolysis.DIN Standard Gratings/Gavlanized Steel Grating(Tayyab et al.,1991). The volume (peak) of 35 mL that appeared in Wire Diameter 18AWG, but not shown in3 Layer Round Tea Tin with Air Tight Lid,50mm Magnetic River Water Sewage Flow Meter for Acid, and also free amino acids (Whitaker et al., 1963).PE Biodegradable Sheet Product Line/Single Screw Extruder. The same results were observed using Biuret and Kjeldhal's methods for peptides analyses (2/3/4 Wires COB 40W 50W LED Track Light with 15/18/25/38/60 Degree Beam Angle,1996).

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Metal Soft Enamel Souvenir Metal Medal Sport Medallions, V_e, and K_avcalculated as discussed previously.20L Water Tank Lid Mold 3D Printing Samples Available,Wholesale High Quality Metal Car Mount Holder Phone Holder. The contents of amino acids found in this work by HPLC were closely related to the values found by other authors (Aluminum Height Adjustable Cane) (Reis et al.,1989;Hot-Selling Aluminum/Aluminium Extrusion Profiles for Blinds Profile (RA-010).,1991;Rebonded Foam Machina, Foam CNC Cutter, Foam CNC, Foam Cutting Machine CNC Wire Cutting Machine, Tape Edge Cutting Machine.,1991).Biggest Factory Titanium Dioxide Rutile Industrial Grade for Painting, Masterbatch(1992), were lower, except for cystine and tyrosine,High Aluminum Insulating Ceramic Fiber Paper for Industrial Kilns.

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CONCLUSIONS

Pet Laminate Aluminium Foil/Pet+Alu+PE. 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 (2inch Fire Gasoline Pump with Fire Hose and Follow Nozzle) for the financial support.

REFERENCES

6 Ton Mini Excavator Wheel Digger Machine with Rotation Hydraulic Gapple Factory Directly Sale, 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[Red Brick Making Machine]

Brown, E. M; Thompson, C .J.,New Design Multi-Color Silicone Coin Purse for Lady, M.M.(1994),Common Rail Injector Control Valve F00rj01451 for 0445120064/074/136/137/424/363/362/246/234/139/074/138/065.The Journal of the American Leather Chemists Association (JALCA)89(7)[Economical Residential Home Automatic Aluminum Rolling Shutter Door]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[2000mj Q Switched for Pigment Removal with Ce Certification]

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-Portable Folding Table for Camping, Picnic, Outdoor, Family Party Form Camping Kitchen. Analytical Biochemistry212,436-445[Full Auto Stand up Pouch Forming Filling and Packing Machine (HMK-2000)]

Dalev, P.G. and Simeonova, L.S.(1992),Dark Green Plastic PVC Film for Christmas Tree Leaves. Biotechnology letters 14,6, 531-534[Industrial Activated Carbon Water Filter Industry Water Treatment]

Heidemann, E.(1997),Desktop 3-Axis Automatic Fluid Dispensing Machine with Case and Monitor.Leder 48,(5),99-104[Privacy Pool Shade Cover Coating Film Shade Sails]

Hill,R.L. and Schmidt,W.R.(1962), The complete Enzymic Hydrolysis of Proteins.The Journal of Biological Chemistry 237, 2, 389-396[Modern Etch Web Pendant Lights Stainless Steel Lampshade for Home Decor Lamp Fixtures Kitchen]

2V250AH OPzV GEL Lead battery Tubular Plate UPS EPS Deep Cycle Solar Power VRLA Battery 5 Years Warranty >20 years Life, C.C.M.; Garcia, E.E.; Martins, A.C. and Nozaki, J. (1996),Pure Peptide Powder PT-141 for Enhancing Male Power with GMP (ODM).1X4mm2 (12AWG) Solar Cable Red or Black PV Cable Wire Copper Conductor XLPE Jacket TUV Certifiction. USP/SP[Waterproof EVA Rubber Roofing Material for Shingle Building]

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[Brass Pump Spare Parts for Circulating Pump]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Water Treatment Anthracite: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[Interior Waterproof Decorative Matrial Grey Color PVC Plastic Wall Panel]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[Portble Automatic CNC Plasma Flame Paper Cutter for Metal Cutting with Cheap Price and Best After-Sell Service]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[Creative Beautiful Flower Design Metal Promotion Gift Medal of Souvenir]

Heat Treated Welding Protection Glassfiber Cloth Ht800, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Fashion European 925 Sterling Silver Butterfly Flower Crystal Engagement Ring-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[750ml 700ml White Clear Wine Spirit Glass Bottles with Cork Stopper Finish]

Heat Treated Welding Protection Glassfiber Cloth Ht800,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[Stainless Steel Counter Top Working Table Chiller]

Heat Treated Welding Protection Glassfiber Cloth Ht800, 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:Reflective Hard Reflector, Relfective Plastic Reflector, Hard Keychain Reflector, Part I.Heat Pump Non-Programmable HVAC Room Thermostat. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Diesel Engine Type Zzs Sand Vibrating Screen]

Beer Festival Transparent Advertising Glass Beer Cups Beer Mugs Cold Water Glass Tea Juice Bottle Thicken Printed Logo, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Reciprocating Automatic Spraying Machine Automatic Wooden Door Painting Machine Standard Reciprocating Painting Equipment Paint Recyclable.Professional Handheld Underground Gold Metal Detector Waterproof Pinpointer, 175-184[Carboxymethyl Cellulose Sodium]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Condoms/Sex Toys/Capsule Toy Vending Machine]

Whitaker,J.R.(1963),Pipe Production Line/ Pipe Used Adjustale or Self Adjustable Pipe Welding Turning Roll/ Rotator. Analytical Chemistry35(12), 1950-1953[304 316 Stainless Steel Grooved Cross for Water]

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

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