Characterization Of Gelatin Research Articles

Extraction and characterization of gelatin from bovine heart

Abstract The objective of this study was to investigate the effects of initial heat extraction time and enzyme concentration during subsequent pepsin digestion on gelatin quality from bovine heart (BH) connective tissue (CT). BH gelatin was extracted from isolated BH CT at 80 °C for 4 or 6 h. Gelatin remaining in the CT residue was extracted with pepsin at either 100 or 200 mg pepsin/g CT to examine the yield and properties of enzyme-extracted gelatin. BH gelatins extracted at 80 °C showed very good gel strength (241–269 g) compared to that subsequently extracted with pepsin (54–96 g) and gelatin yield increased with the length of heat extraction (3%) although gel strength was reduced. Heat-extracted BH gelatin regardless of duration of heat extraction contained α1 and α2 chain as main components with some degradation peptides. The frequency sweep test of BH gelatins showed independency in a wide range of frequency (1–196 Hz) regardless of whether extracted by heat or pepsin. Heat-extracted BH gelatins exhibited the highest storage modulus values (1165–1245 pa). BH gelatins extracted with the lowest concentration of pepsin showed a higher mean storage modulus value than gelatin extracted with the highest pepsin concentration. The present study is the first to demonstrate that gelatin extracted from BH collagen using heat and pepsin has characteristics comparable to other sources of gelatin, making it suitable for many applications, and confirms that the use of pepsin increases BH gelatin yield with a concomitant reduction in gelatin quality.

Extraction and characterization of gelatin from Lates niloticus and potential industrial applications

Kiplagat CS, Onyari JM, Mulaa F, Wabomba J. 2018. Extraction and characterization of gelatin from Lates niloticus and potential industrial applications. Biofarmasi J Nat Prod Biochem 16: 53-64. This research aims to extract and characterize gelatin from Lates niloticus (Nile perch) scales, then blend it with polyvinyl alcohol (PVA). Hydrolysis of the scales was done using a crude alkaline protease harvested from a bacterium, Bacillus cereus strain wwcp 1, obtained from Lake Bogoria. The lyophilized solution yielded 16.3% of gelatin powder calculated from the dry weight of the scales. The sample was characterized using infrared spectroscopy and showed peaks at 3442 cm-1, 1653 cm-1 and ~ 1590 cm-1 corresponding to Amide A, Amide I and Amide II bands respectively. The amino acid analysis shows that glycine was the most abundant amino acid (21.7%), followed by proline (14.6%) and alanine (11.8%). Isoleucine, Histidine, and Tyrosine were the least abundant (1.8, 1.4 and 0.9% respectively). Polyvinyl alcohol-gelatin blend films of various compositions ranging from 10% to 90% PVA were prepared by solution casting method. Differential Scanning Calorimetry (DSC) and Thermo-gravimetric Analysis (TGA) tests showed the films had glass transition, melting and thermal decomposition onset temperatures intermediate between those of the respective individual polymers ( PVA and gelatin). The thermal stability of the films reduced with the increase in the amount of the less thermally stable constituent. Lastly, potential applications of the prepared blend films were investigated. Batch experiments to assess the potential of the polymer blend films as an adsorbent material was done using Methylene Blue dye. The films were found to adsorb up to 64% of the dye and the percentage of dye removal varied with the initial concentration of the dye and contact time.

Extraction and characterization of gelatin from the feet of Pekin duck (Anas platyrhynchos domestica) as affected by acid, alkaline, and enzyme pretreatment

The effects of different pretreatments on yield and composition of extraction, physicochemical, and rheological properties of duck feet gelatin (DFG) were investigated. Gelatins were extracted from the whole feet of Pekin duck with an average yield of 4.09%, 3.65%, and 5.75% for acidic (Ac-DFG), alkaline (Al-DFG), and enzymatic (En-DFG) pretreatment on a wet weight basis, respectively. Proteins at 81.38%, 79.41%, 82.55%, and 87.38% were the major composition for Ac-DFG, Al-DFG, En-DFG, and bovine, respectively. Amino acid analysis showed glycine as the predominant amino acid in Ac-DFG, followed by hydroxyproline, proline, and alanine for Ac-DFG, Al-DFG, and En-DFG, respectively. Rheological analysis indicated that the maximum elastic modulus (9972.25Pa) and loss modulus (4956.28Pa) for Ac-DFG gelatin were significantly higher than those of other gelatins. Extracted gelatins contained α1 and α2 chains as the predominant components, and enzymatic gelatin had low molecular weight peptides. Fourier transform infrared spectroscopy showed that the peak of the gelatins was mainly positioned in the amide band region (amides I, II, and III). A considerable loss of molecular-order triple helical structure was also observed after pepsin treatment. In summary, duck feet gelatin has potential to replace as mammalian gelatin in food and pharmaceutical industry.

Extraction and Characterization of Gelatin from Bovine Heart

Extraction and Characterization of Gelatin from Bovine Heart

Extraction and Characterization of Gelatin from Bovine Lung

Gelatin is extracted from animal tissues using heat usually with low yields, but pepsin may increase high quality gelatin yield per unit of tissue. Gelatin from bovine lungs was extracted using heat and pepsin and the resulting gelatins were characterized. Pepsin increased gelatin yield by about 9-fold that of heat extraction alone. All bovine lung gelatin contained protein as the major proximate component, with little ash and non-detectable fat. Bovine lung gelatin had pH, moisture and protein comparable to or less than that of commercial bovine gelatin and decreased ash. Transmittance of bovine lung gelatin was substantially reduced compared to that of commercial bovine gelatin but had increased water and fat-binding capacity, and comparable or increased gelling and melting temperature. Gel strengths of bovine lung gelatin were comparable to or lower than and foam stability and emulsifying activity were lower than commercial bovine gelatin. Increased imino acid (proline and hydroxyproline) content was associated with increased gelling and melting temperatures and was comparable to commercial bovine gelatin. Heat-extracted bovine lung gelatin contained predominantly collagen γ– chains, β– chains and α– chains (α1(I) and α2(I)), with some low molecular weight peptides, while the pepsin-extracted lung gelatins were characterized by comparatively decreased β– and α– chains and increased low molecular weight peptides. The gel strength of heat-extracted bovine lung gelatin was higher than that of pepsin-extracted gelatins, indicating that additional yield was associated with reduced gelatin quality. Bovine lung is a potential source of gelatin for application in diversified industrial fields and use of pepsin is a viable method for extracting additional gelatin after heat extraction of high quality (increased gel strength) gelatin from bovine lung.

Poultry Processing Waste as an Alternative Source for Mammalian Gelatin: Extraction and Characterization of Gelatin from Chicken Feet Using Food Grade Acids

The present study was focused on extraction of gelatin from chicken feet using different food grade acids (acetic-, citric- and lactic-acid) at different concentrations. The gelatin thus obtained was evaluated for its physico-chemical properties. Gelatin yields in various acid treatments ranged between 6.18 and 14.47% on wet weight basis. The protein content of chicken feet gelatin (CFG) obtained with different acid treatments was in the range of 80.53–90.20%; while, the pH was in the range of 3.43–5.23. Among all, CFG extracted with 1.5% (v/v) acetic acid showed the highest bloom strength (204.3 g) which was similar to commercial porcine gelatin. The CFG prepared using 4.5% (v/v) citric acid had a bloom value of 165.8 g. The bloom strength of CFG prepared in our study was relatively higher than fish gelatin and slightly lower than porcine gelatin. Hydroxyproline content (g/100 g) of CFG prepared using different acids was in the range of 4.58–9.5. The CFG contained glycine (~30%), imino acids (proline + hydroxyl proline; ~25%) and alanine (~9%) as the major amino acids. CFG, irrespective of the acids used for preparation, exhibited moderate to strong emulsifying and foaming capacities. The results clearly indicate the potential of CFG to be an effective alternative to mammalian gelatin.

Extraction and Characterization of Gelatin from the Head Waste of Tiger Tooth Croaker (Otolithes ruber)

Purpose The objective of the present study was to valorize the tiger tooth croaker (Otolithes ruber) fish head waste for the extraction of high value protein based component ‘gelatin’.

Extraction and characterization of gelatin from two edible Sudanese insects and its applications in ice cream making.

Three methods were used for extraction of gelatin from two insects, melon bug (Coridius viduatus) and sorghum bug (Agonoscelis versicoloratus versicoloratus). Extraction of insect gelatin using hot water gave higher yield reached up to 3.0%, followed by mild acid extraction which gave 1.5% and distilled water extraction which gave only 1.0%, respectively. The obtained gelatins were characterized by FTIR and the spectra of insect's gelatin seem to be similar when compared with commercial gelatin. Amide II bands of gelatins from melon and sorghum bug appeared around at 1542-1537 cm(-1). Slight differences in the amino acid composition of gelatin extracted from the two insects were observed. Ice cream was made by using 0.5% insect's gelatin and compared with that made using 0.5% commercial gelatin as stabilizing agent. The properties of the obtained ice cream produced using insects gelatin were significantly different when compared with that made using commercial gelatin.

Preparation and characterization of gelatin from collagen biomass obtained through a pH-shifting process of mechanically separated turkey meat

Gelatins were extracted from mechanically separated turkey meat following 2 different approaches. The first method was based on a 2-stage batch extraction at 50 and 60°C, respectively, whereas the second method consisted of recovering gelatin from a collagen biomass obtained during a pH-shifting process. The yield of gelatin produced by the latter method was twice that obtained by the former method (13.51 and 6.36%, respectively). The chemical composition, as well as the rheological and the functional properties, of all extracted gelatins were evaluated. Gelatin recovered from the collagen biomass had higher molecular weight components and significantly greater (P < 0.05) bloom value (353.2 g) compared with thermally extracted gelatins. However, gelatin extracted at 60°C possessed higher (P < 0.05) foaming properties, as well as better emulsifying activity, than gelatin extracted from the 50°C treatment and the collagen biomass. The present study revealed that high-quality gelatins can be prepared from mechanically separated turkey meat through precipitation and thermal solubilization of collagen biomass obtained during a pH-shifting process.

Gelatin- and hydroxyapatite-based cryogels for bone tissue engineering: synthesis, characterization,in vitroandin vivobiocompatibility

The aim of this study was the synthesis and characterization of gelatin- and hydroxyapatite (osteoconductive component of bone)-based cryogels for tissue-engineering applications. Preliminary in vitro and in vivo biocompatibility tests were conducted. Gelatin- and hydroxyapatite-based cryogels of varying concentrations were synthesized using glutaraldehyde as the crosslinking agent. Chemical structure, pore morphology, pore size distribution, mechanical properties, swelling characteristics and degradation profiles of the synthesized cryogels were demonstrated by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), mercury porosimetry, a mechanical test device, swelling ratio tests and weight loss measurements, respectively. In vitro cell viability and in vivo biocompatility tests were performed in order to show the performance of the cryogels in the biological environment. Changing the concentrations of gelatin, hydroxyapatite and crosslinker changed the chemical structure, pore size and pore size distribution of the cryogels, which in turn resulted in the ultimate behaviour (mechanical properties, swelling ratio, degradation profile). In vitro cell culture tests showed the viability of the cells. The cryogels did not show any cytotoxic effects on the cells. Clinical outcomes and the gross pathological results demonstrated that there was no necrosis noted in the abdominal and thoracic regions at the end of implantation and the implanted cryogel was found to be non-irritant and non-toxic at 12 weeks of implantation. Copyright © 2013 John Wiley & Sons, Ltd.

Development and characterization of gelatin and ethylcellulose microparticles designed as platforms to delivery fluoride

Purpose: To develop and characterize microparticles containing fluoride sources (FS) from sodium fluoride, sodium monofluorophosphate (MFP) or aminofluoride and evaluate their characteristics as fluoride delivery systems.Methods: Ethylcellulose microparticles containing fluoride (EM) were prepared by emulsification of ethyl acetate dispersion containing polymer and FS (ethylcellulose:FS ratio of 1:0.25 wt/wt) with aqueous external phase containing polysorbate 80 (0.8% vol/vol) using the volume ratio (organic:aqueous) of 1:5. The organic solvent was evaporated; microparticles were collected by centrifuging, washed with deionized water and freeze-dried. Gelatin microparticles containing FS (GM) was obtained by dispersion of the natural polymer in water, adding FS (6:1 wt/wt) and 20% (wt/wt) of mannitol. The final dispersions were spray-dried. Particle morphology and size were investigated using optical microscopy. The content of fluoride ions in the microparticles was quantified using a potentiometric method. The encapsulation efficiency and in vitro release profile of fluoride was also determined.Results: Microparticles exhibited polydispersity and mean diameters <145.35 and <124.22 µm for EM and GM, respectively. Considering the entrapment efficiency, the spray-drying technique exhibited greater values than microencapsulation by emulsification and solvent evaporation. The release profile of fluoride ions from microparticles was shown to be modified, fitted first order and guided by Fickian diffusion.Conclusions: Microparticles prepared with ethylcellulose or gelatin can be used as platform for oral delivery of fluoride, providing a means to increase the local supply of this ion in a controlled manner, providing an increased protection against caries. Moreover, further investigations are needed to demonstrate this property in vivo.

Preparation and Characterization of Gelatin–Poly(L-lactic) Acid/Poly(hydroxybutyrate-co-hydroxyvalerate) Composite Nanofibrous Scaffolds

Poly(L-lactic) acid (PLLA) scaffolds, prepared by electrospinning technology, have been suggested for use in tissue engineering. They remain a challenge for application in biological fields due to PLLA's slow degradation and hydrophobic nature. We describe PLLA, PLLA/poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV), and PLLA/PHBV/gelatin (Gt) composite nanofiberous scaffolds (Gt–PLLA/PHBV) electrospun by changing the electrospinning technology. The morphologies and hydrophilicity of these fibers were characterized by scanning electron microscopy (SEM) and water contact angle measurement. The results showed that the addition of PHBV and Gt resulted in a decrease in the diameters and their distribution and greatly improved the hydrophilicity. The in-vitro degradation test indicated that GT–PLLA/PHBV composite scaffolds exhibited a faster degradation rate than PLLA and PLLA/PHBV scaffolds. Dermal fibroblasts viabilities on nanofibrous scaffolds were characterized by [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] (MTT) assay and cell morphologies after 7 days culture. Results indicated that the GT–PLLA/PHBV composite nanofibers showed the highest bioactivity among the three scaffolds and increased with increasing time. The SEM images of cells/scaffolds composite materials showed the GT–PLLA/PHBV composite nanofibers enhanced the dermal fibroblasts's adhesion, proliferation, and spreading. It is suggested that the nanofibrous composite scaffolds of GT–PLLA/PHBV composites would be a promising candidate for tissue engineering scaffolds.

Physicochemical Properties of Mammalian Gelatin in Relation to Membrane Process Requirement

In any of the membrane process application, understanding of the characteristics of the feed solution is essential in order to achieve desired level of separation performance. In this study, in an effort to substitute evaporation with membrane processes partially, experiments were carried out to investigate the physicochemical properties of gelatins, namely, molecular weight distribution, pH, viscosity, isoelectric point, and gel strength, which are, of foremost, important parameters in the characterization of gelatin. Two different mammalian gelatins, i.e. from bovine (type B) and porcine (type A) sources, were used in this study. The pH was significantly varied for all gelatins in the vicinity of 4.75–5.51 (±0.01). Experimental result revealed that both sources of mammalian gelatin contained components of different molecular weights with wide distribution ranging from 10 to 400 kDa. Analysis of the molecular weight distribution result also showed strong correlation between average molecular weight and gel strength of gelatin. The isoelectric points of gelatins from bovine were 4.60 ± 0.08 to 5.25 ± 0.43 and porcine gelatins were in the range of 7–9.3, which agreed well with the results obtained from other researchers. The high bloom strength mammalian gelatins were also significantly more viscous and thus, had a higher melting point.

Processing optimization and characterization of gelatin from lizardfish (Saurida spp.) scales

Abstract Response surface methodology (RSM) with a 4-factor, 5-level central composite design (CCD) was conducted to ascertain the optimum gelatin extraction conditions from lizardfish scales. The effects of concentration of NaOH (%, X 1 ), treatment time (h, X 2 ), extraction temperature (°C, X 3 ) and extraction time (h, X 4 ) were determined. The responses included extraction yield (%), gel strength (g) at 9–10 °C and viscosity (cP) at 25 °C. The results showed the optimum conditions for the highest values of the three responses when a concentration of NaOH at 0.51%, a treatment time at 3.10 h, an extraction temperature at 78.5 °C and an extraction time at 3.02 h. The predicted responses were 10.7% extraction yield, 240 g gel strength and 5.61 cP viscosity. The experimental values were 10.6 ± 0.82% extraction yield, 252 ± 1.21 g gel strength and 7.50 ± 0.28 cP viscosity. The physicochemical properties of the lizardfish scales gelatin were characterized and the results indicated high protein and low ash content. Texture profile analysis (TPA) with compression was carried out at 30% deformation. The lizardfish scales gelatin was found to contain 20.4% imino acids (proline and hydroxyproline). Furthermore, slightly loose strands of the gel microstructure were observed using scanning electron microscopy (SEM).

Synthesis and characterisation of gelatin–nano hydroxyapatite composite scaffolds for bone tissue engineering

Gelatin–hydroxyapatite (HAp) nanocomposites have been prepared by particulate leaching technique using glutaraldehyde (GTA) as cross-linking agent for polymer. The porosity in the scaffolds was controlled using sodium chloride as porogen agent. Microstructural investigation by scanning electron microscopy (SEM), revealed the formation of a well interconnected porous scaffold with pore size in the range of 100–200 μm. X-ray diffraction and Fourier transform infrared spectroscopy were used to confirm the formation of crystalline HAp as well the presence of both constituents in the composite samples. The bioactivity of the samples was evaluated by conducting MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and cell adhesion tests. The results suggest that the use of GTA in excess of 0˙25% can be detrimental to cell survival. Cell attachment on the nanocomposite scaffold was verified by microscopic analysis.

Comparison of Selective Attachment and Growth of Smooth Muscle Cells on Gelatin- and Fibronectin-Coated Micropatterns

Tissue engineering research has been on going for many years, people are making all the effort to explore the cell functions in cellular level and even in molecular level. Making the cells functional in an in vitro environment is a preliminary goal for the implantation and repair of complicated tissues/organs. Fabricating artificial ECM to mimic the in vivo environment is an essential approach in tissue engineering. The work in this paper is to study how rat aorta smooth muscle cells (RASMCs) behave in two engineered cell culture scaffolds: gelatin- and fibronectin (FN)-coated micropatterns. The investigation on the initial attachment and further growth of SMCs cultured on gelatin- and FN-coated micropatterns was addressed. This study focused on both the characterization of gelatin and fibronectin assembly properties and cell responses to these two protein-coated micropatterns. Thin film patterns with gelatin and fibronectin coatings were fabricated on microscope glass slides using photolithography, electrostatic layer-by-layer self-assembly and lift-off (LbL-LO) technologies. In this work, the scaffolds were built up by commonly used polyelectrolyte materials and proteins through LbL process, containing cationic poly(diallyldimethylammonium chloride) (PDDA), poly(allylamine hydrochloride) (PAH), anionic poly(sodium 4-styrenesulfonate) (PSS), gelatin and fibronectin. The resulting polyelectrolyte thin films were characterized by contact angle (CA), quartz crystal microbalance (QCM), atomic force microscopy (AFM), and fluorescence microscopy. CA measurement shows the consistent hydrophylicity of gelatin surfaces in different number of layers with LbL deposition method. Different from our previous QCM measurement of gelatin, fibronectin does not show high electrostatic attraction to either positively or negatively charged polyelectrolytes, although it can be weakly assembled to both polyelectrolyte surfaces. AFM images show Gelatin- and FN-coated micropatterns are around 50-60 nm thick. RASMCs were cultured on these gelatin- and FN-coated micropatterns. It was observed that, for the cells cultured on gelatin-coated micropatterns, they initially landed on the gelatin-coated surface, not on the PDDA-coated surface in between. But further growth of the cells was affected by the shape of the patterns: strip pattern limited cell growth beyond the patterns, but square patterns could not. While, it was found interestingly, for the cells cultured on FN-coated micropatterns, SMCs initially landed on PDDA-coated surface, and then migrated to FN-coated both square and strip patterns. These findings indicate that both gelatin and fibronectin are adhesive proteins, but they have different effects on the initial attachment and later growth for SMCs.

Laser light‐scattering characterization of gelatin in formamide

AbstractLaser light scattering (LLS) including angular dependence of absolute integrated scattered intensity (static LLS) and of the spectral distribution (dynamic LLS) has been used successfully to characterize gelatin in formamide at room temperature. In static LLS, the use of formamide as a single solvent instead of an aqueous salt solution avoids the well‐known problem of preferential sorption of salts in the domain of gelatin molecules. Therefore the true weight‐average molecular weight Mw, the z‐average radius of gyration, and the second virial coefficient have been determined. In dynamic LLS, precise measurements of the intensity‐intensity time correlation function permit a Laplace inversion to obtain an estimate of the normalized characteristic linewidth distribution which could be reduced to a translational diffusion coefficient distribution, G(D). This report shows that the calibration between D and M can be established from Mw and G(D) by using only two broadly distributed gelatins instead of a set of narrowly distributed gelatin standards. After establishing a calibration between D and M, we were able to estimate the molecular weight distribution of gelatin from G(D). © 1994 John Wiley &amp; Sons, Inc.

Isolation of Gelatin from Ancient Bones

The isolation and characterization of gelatin from 12,000-year-old deer antlers is described. Use of gelatin from ancient bones for carbon-14 dating may improve the accuracy of the dating procedure because gelatin is not likely to be contaminated by extraneous carbon.

Isolation of gelatin from ancient bones.

The isolation and characterization of gelatin from 12,000-year-old deer antlers is described. Use of gelatin from ancient bones for carbon-14 dating may improve the accuracy of the dating procedure because gelatin is not likely to be contaminated by extraneous carbon.