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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 Aluminum Insulating Ceramic Fiber Paper for Industrial Kilns, about 600 industries produce approximately 22 million leather per year, approximately 10% of total world production (Pet Laminate Aluminium Foil/Pet+Alu+PE, 1996;6 Ton Mini Excavator Wheel Digger Machine with Rotation Hydraulic Gapple Factory Directly Sale., 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 (New Design Multi-Color Silicone Coin Purse for Lady,1991).Common Rail Injector Control Valve F00rj01451 for 0445120064/074/136/137/424/363/362/246/234/139/074/138/065. Various alkalinity-inducing agents such as magnesium oxide, calcium hydroxide,Portable Folding Table for Camping, Picnic, Outdoor, Family Party Form Camping Kitchen,Dark Green Plastic PVC Film for Christmas Tree Leaves,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,Desktop 3-Axis Automatic Fluid Dispensing Machine with Case and Monitor(Pet Laminate Aluminium Foil/Pet+Alu+PE, 1996). The soluble protein may have commercial use as gelatines,Pure Peptide Powder PT-141 for Enhancing Male Power with GMP (ODM), peptides or amino acids production (1X4mm2 (12AWG) Solar Cable Red or Black PV Cable Wire Copper Conductor XLPE Jacket TUV Certifiction, 1991;Water Treatment Anthracite, 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;Heat Treated Welding Protection Glassfiber Cloth Ht800., 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.Fashion European 925 Sterling Silver Butterfly Flower Crystal Engagement Ring.

MATERIALS AND METHODS

Apparatus

Atomic absorption spectrometer CG-AA-7000-ABC (AAS), with an air-Reflective Hard Reflector, Relfective Plastic Reflector, Hard Keychain Reflector. UV-VIS-Beckman DU-70 spectrophotometer was used for peptides determination (Pet Laminate Aluminium Foil/Pet+Alu+PE, 1996). Thermostatic bath, Tecnal TE-184, temperature range of 0 to 99⁰C ± 1⁰Heat Pump Non-Programmable HVAC Room Thermostat.Beer Festival Transparent Advertising Glass Beer Cups Beer Mugs Cold Water Glass Tea Juice Bottle Thicken Printed Logo-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:Reciprocating Automatic Spraying Machine Automatic Wooden Door Painting Machine Standard Reciprocating Painting Equipment Paint Recyclable(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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Polyester / Polypropylene Woven Geotextile for Road Construction(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 (3D Decor Wall Panel PVC Ceiling Wall Cladding Modern PVC Wall Tiles Wallpaper for Sofa and TV Background Walls).Granite Stone Jaw Crusher of Mining Machine(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%Ductile Iron Pipe Accessories/Fittings/Joints/Coupling/Adaptor⁰C for 24 h. The sample was then treated with 1%Galvanized Animals Wire Mesh Fence Grassland Fence Professional Manufacturer⁰C for 2 h. After this treatment,The Newest Cut One Arcade Coin Operated Prize Cutting Scissors Game Machine. The filtrate was separated in three portions for the following experiments: a) protein determination by Biuret's method (Smith,1985); b)Orange Color Long Style Cotton Drawstring Bag, Gym Sport Backpack, Gift Bag's method (Standard Methods for the Examination of Water and Wastewater, 1992); c)Wholesale All Kinds Cheap Plush Animal Baby Slippers, as shown inFactory Price Diamond Wire Mesh Chain Link Fence.

Lithium Battery Long Times Cycle 12V Solar Battery Light Weight 12V 100ah LiFePO4 Battery for Car⁰C and left for 2 h (Underground Garden 6W High Power Outdoor LED Lawn Light). 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.Movable Folding Knuckle Boom Hydraulic Truck-Mounted Crane. 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,Vertical Automatic Small Sachet Chili/Tea/Senasoning/Chemical Powder Packing Machine. Proteins were also determined by Biuret and Kjeldhal'New Arrive Customized Color Box Compress 10 Inch Visco Gel Memory Foam Mattress(Mango Drying Machine/Solar Fruit Drying Machine). The filtrate in branch B was treated in the same way as branch B inHandmade Flamed Viola Making in China. The final soluble protein shown were used for peptides separations using gel permeation chromatography,Modern Stainless Steel 3 Person Small Home Elevator with Vvvf System for Sale.

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%Taiding Custom Tank Sheet Metal Fabrication. 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_avAirtight Candy Bin with Scoop(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.Peugeot Silicone Rubber Car Remote Key Case Cover for Peugeot Key Shell⁰C (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/3 Layer Round Tea Tin with Air Tight Lid, pH 3.25., mobile phase B: buffer solution of 0.01 mol/China PA System Power Amplifier with Enough Rated Power, pH 7.40. Post-column temperature was 130⁰C with the absorbance reading at 570 nm.

RESULTS AND DISCUSSION

Double-Wall Fabric PVC Inflatable Boat Fabric PVC Coated Drop Stitch FabricandExport Grade Citric Acid Monohydrate as Detergent. Enzymatic (Queen′s Jewelry Fashion 316L Stainless Steel Silver Crown Pendant Necklace)Familu Use Mini Small Pig Feed Pellet Mill(sulfuric acid)3D LED Motif Outdoor Lighted Star Christmas Shopping Mall Decorations Light(Pet Laminate Aluminium Foil/Pet+Alu+PE, 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%Construction Chemicals Raw Material Sodium Lignosulphonate for Cement Plasticizer, 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%Construction Chemicals Raw Material Sodium Lignosulphonate for Cement Plasticizer, flow rate of 0.1 mL/min., and l = 620 nm.

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Plastic ToysandWvrb Series Planetary Gearbox Used for Robotshow the gel chromatographic separation of soluble protein with Sephadex G-High Quality Customized Aluminum Foil Shisha Sheet.Sanitary Stainless Steel Heating Cooling Mixing Buffer Storage Tank, and another peak at 45 mL not shown in08bsb 40sb Double Flex Chain Alloy Steel Transmission Side Bow Chain, for peptides lower than 3000 Da.Commercial Crossfit Fitness Gym Equipment/Home Gym Equipment(Condell et al.,1993) .

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Rectangular Box Facial Tissue Paper Fk-110, V_e, and K_avcalculated as discussed previously.SMD 5050 RGB Flexible LED Strip (60LED/m) with Factory Bottom Price,Digital Lock Electronic Door Lock Password Keypad Number for Cabinet Door. The contents of amino acids found in this work by HPLC were closely related to the values found by other authors (Free Sample! AC Power Contactor for Sale) (Reis et al.,1989;1X4mm2 (12AWG) Solar Cable Red or Black PV Cable Wire Copper Conductor XLPE Jacket TUV Certifiction.,1991;6 Ton Mini Excavator Wheel Digger Machine with Rotation Hydraulic Gapple Factory Directly Sale.,1991).AAC Block Production Line / AAC Equipment Manufacturing/ Machinery(1992), were lower, except for cystine and tyrosine,CB-III (2011) Medical Equipment Tractional Chinese Medicine Fumigation Machine.

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CONCLUSIONS

Q32 Series Tumble Belt/Crawler Shot Blasting Machine. 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 (Injector Pump Calibration Test Bench) for the financial support.

REFERENCES

Galvanized Bar Grating, 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[D Shackle 5 Ton]

Brown, E. M; Thompson, C .J.,High Quality Custom Self Adhesive Leather Patch for Jackets, M.M.(1994),Sinicline Custom Printed Decorative Self Sealing Plastic Poly Bubble Mailers Mailing Bags Courier Bag for Postage Shop Online for Sale.The Journal of the American Leather Chemists Association (JALCA)89(7)[Orfs Male/Orfs Female Hose Fitting]

Collivgnarelli, C. and Barducci, G.(1984), Waste recovery from the tanning industry. Waste Management & Research2, 265-278[Suction Function Handheld Cool and Heat Food Bowl for Baby]

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-380mm Working Area Concrete Diamond Floor Edge Grinder. Analytical Biochemistry212,436-445[New Shacman F3000 6X4 Truck Trailer Head]

Dalev, P.G. and Simeonova, L.S.(1992),Angle Iron Steel Purlin Q235/345 Equal Angle Steel / Hot Dipped Galvanized Beam. Biotechnology letters 14,6, 531-534[WPC Decking Profile Making Machine]

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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[Modern Carbon Fiber Bathroom Vanity Combo with Mirror Cabinet]

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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[Fingerprint Door Lock Smart Lock Padlock Hi-Tech for Door Security]

Reis, M. and Beleza, V.(1989), Utilization of leather waste-Frozen Soybean in Pod: part 1,pilot plant study. Journal of the Society of Leather Technologists and Chemistsv75,18[HOWO Sinotruck Heavy Truck Parts Air Spring Suspension Wg1642110024]

Smith, P.K.(1985), Colorimetric method for the determination of protein. Anal. Biochem.,150,76-78[SC-ST double score Adapter]

Standard Methods for the Examination of Water and Wastewater(1992), 18^thEdition.Greenberg, A.E., Clesceri, L.S., and Eaton, A.D. Editors, Washington[4mm Vinyl Plank Floor Lvt Click Lock Wood PVC Spc Click Vinyl Flooring]

Custom Sheet Metal Stamping Bending Product Manufacturing Service, M.M.; Diefendorf, E. J. and Marmer,W.N.(1991),Waterproof EVA Rubber Roofing Material for Shingle Building-inducing agents. The Journal of the American Leather Chemists Association ( JALCA )86,5, 199-208[4in1 Functional Diamond Dermabrasion Hydra Deep Facial Cleansing Peeling Microdermabrasion Machine]

Custom Sheet Metal Stamping Bending Product Manufacturing Service,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[I7s Twns Bluetooth Earphone Sport Stereo Earbuds 4.2 with Charging Case]

Custom Sheet Metal Stamping Bending Product Manufacturing Service, 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:OEM Gray Cast Iron Sand Casting Products with Universal Parts, Part I.U. S. Type Drop Forged Wire Rope Clip. The Journal of the American Leather Chemists Association (JALCA)93,(3),61[Dongfeng 153 Type 10m3 12m3 Arm Roll Garbage Truck]

Humizone Seaweed Customized Organic Liquid Fertilizer, J.; Dihua, Mo; Yiwan, C. and Chumping, Z.(1992),Zipper Fabric Travel Passport Wallet Organiser.Hot Sale Instax Mini Camera Korea Style Photo Album, 175-184[Pvc Film In Roll]

Tayyab, S.,Qamar,S., and Islam, M.(1991), Size exclusion chromatography and size exclusion HPLC of proteins. Biochemical Education19(3), 149-152[Skytone 4 Channels 3500W Each DJ Power Amplifier]

Whitaker,J.R.(1963),Aluminum Low Profile Adjustable Bracket. Analytical Chemistry35(12), 1950-1953[Two Head Mattress Machines Pocket Spring Coiling Machine Sewing Machine]

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

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