Actinidin was used to pretreat the bovine hide and ultrasonic wave (53 kHz and 500 W) was used for the time durations of 2, 4 and 6 h at 60 °C to extract gelatin samples (UA2, UA4 and UA6, respectively). Control (UAC) gelatin was extracted using ultrasound for 6 h at 60 °C without enzyme pretreatment. There was significant (p < 0.05) increase in gelatin yield as the time duration of ultrasound treatment increased with UA6 giving the highest yield of 19.65%. Gel strength and viscosity of UAC and UA6 extracted gelatin samples were 627.53 and 502.16 g and 16.33 and 15.60 mPa.s, respectively. Longer duration of ultrasound treatment increased amino acids content of the extracted gelatin and UAC exhibited the highest content of amino acids. Progressive degradation of polypeptide chains was observed in the protein pattern of the extracted gelatin as the time duration of ultrasound extraction increased. Fourier transform infrared (FTIR) spectroscopy depicted loss of molecular order and degradation in UA6. Scanning electron microscopy (SEM) revealed protein aggregation and network formation in the gelatin samples with increasing time of ultrasound treatment. The study indicated that ultrasound assisted gelatin extraction using actinidin exhibited high yield with good quality gelatin.
Corn starches with different amylose-to-amylopectin ratio (waxy, normal, Hylon V, and Hylon VII) were treated with five doses of gamma irradiation (1, 5, 10, 25, and 50 kGy). The effects of gamma irradiation on the physicochemical properties of starch samples were investigated. Waxy samples showed an increase of amylose-like fractions when irradiated at 10 kGy. The reduction in apparent amylose content increased with amylose content when underwent irradiation at 25 and 50 kGy. Low amylose starches lost their pasting ability when irradiated at 25 and 50 kGy. Results from thermal behavior and pasting profile suggested that low level of cross-linking occurred in Hylon VII samples irradiated at 5 kGy. Severe reduction in pasting properties, gelatinization temperatures and relative crystallinity with increasing irradiation intensity revealed that waxy samples were affected more by gamma irradiation; this also indicated amylopectin was the starch fraction most affected by gamma irradiation. Alteration level was portrayed differently when different kind of physicochemical properties were investigated, in which the pasting properties and crystallinity of starches were more immensely influenced by gamma irradiation while thermal behavior was less affected. Despite the irradiation level, the morphology and crystal pattern of starch granules were found remain unchanged by irradiation.
Poor quality and quantity of DNA extracted from gelatin and gelatin capsules often causes failure in the determination of animal species using PCR. Gelatin, which is mainly derived from porcine and bovine, has been a matter of concern among customers in order to fulfill religious obligation and safety precaution against several transmissible infectious diseases associated with bovine species. Thus, optimised DNA extraction from gelatin is very important for successful real-time PCR detection of gelatin species. In this work, the DNA extraction method was optimised in terms of lysis incubation period and inclusion of pre-treatment pH modification of samples.
Interventional radiology procedures are becoming more challenging over time; thus, there is a need for excellent and reliable training methods. Training on live patients is neither safe nor an ethical solution. Alternatives are many and varied, but the most popular is ultrasound guided simulators. This report shows how a simple, homemade, low-cost phantom material, and construction modules can provide several advantages over ordinary gelatin phantoms. A new layering technique and target synthesis are described for the biopsy phantom, including tips on decreasing the needle pass artifact as well as controlling the mixture echogenicity.
High invasive cancer cells are thought to recruit specialised actin-rich protrusions for invasion in metastasis process. These protrusions are termed invadopodia. To study invadopodia formation, one of the first challenges faced by researchers has been to optimise the cell line passage number in order to be used for the invadopodia assay. Therefore, this study aims to investigate the effects of the passage number on invadopodia formation in MDA-MB-231 breast cancer cell line. Invadopodia assay was used to achieve the aim of the study. The results provided evidence that invadopodia formation is affected by the high passage number. The cells were also tested with dimethyloxalylglycine (DMOG) a hypoxic mimicking agent which is known to be an invadopodia inducer, the results showed that the cells in low passage number (P7) treated with DMOG increase the cells forming invadopodia, while the cells with high passage number (P35) showed that DMOG fails to stimulate the cells to form invadopodia. Furthermore, the cells with high passage number after passage 15 are starting to lose the ability to degrade the gelatin. In conclusion, this study suggests that only cells with a low passage number, less than passage 15 should be used in the study of invadopodia formation to obtain the results in the search for molecular targets and signaling at invadopodia.
Gelatins from the skin of four local marine fish, namely “kerapu” (Epinephelus sexfasciatus), “jenahak” (Lutjianus argentimaculatus), “kembung” (Rastrelliger kanagurta), and “kerisi” (Pristipomodes typus) have been successfully extracted by acid extraction. Results characterization showed that the fish gelatins were comparable to the fish gelatins from other fish species previously reported. They appeared snowy white in color with crystal-like and light texture. The gelatine extracted from “kerapu” had the strongest fishy odor, followed by the gelatines derived from “jenahak”, “kembung” and “kerisi”. In terms of bloom strength, the gelatin extracted from “kerapu” was found to be the strongest one compared to others, with the bloom value of more than 2000 g. The gelatins developed in this study contained almost all essential amino acids, with glycine being the most predominant one.
The peptide composition of gelatin is known to vary very common that the electrophoretic pattern of gelatin from one source differs from another source even for the same raw material. Therefore, the present study aimed to use proteomics field to identify gelatin polypeptides biomarker for depending on the condition under which collagen is hydrolyzed. Hence, it is porcine skins. The polypeptides obtained for porcine skin gelatins can be used as reference in future to detect the origins of gelatin added in the processed food. We compared porcine skin gelatin samples obtained from three producers. Total average numbers of polypeptides of porcine skin gelatins from company A, B and C were 303 ± 2.8, 285.5 ± 3.5 and 270.5 ± 4.9 spots respectively. 10 biomarkers were identified and presented in all different companies. We also did a mixture of porcine and bovine skin gelatin to detect the presence of these 10 biomarkers. The level of adulteration that could be detected was as low as 1.0% w/w
The study was conducted to detect the porcine DNA in pharmaceutical products in local market using polymerase chain reaction (PCR) and southern-hybridization on the biochip. A total of 113 (n=113) of hard (82 samples) and soft gel (31 samples) capsules from pharmaceutical products were purchased and tested for the presence of porcine DNA for Halal authentication. All capsules were gelatin-based purchased from local over the counter (OTC) markets. Of all samples tested, 37.2% (42/113) contained porcine DNA. While, none porcine DNA band was detected for 62.8% (71/113) of capsules tested. All samples which were positive toward porcine DNA were imported pharmaceutical products with none Halal logo. Results in the presence study demonstrated that the PCR techniques and southern-hybridization on the biochip is suitable tool for monitoring the Haram component in highly processed product of soft and hard capsule.
This study investigated the antioxidant activity and functional properties of fractionated cobia skin gelatin hydrolysate (CSGH) at different molecular weights (10, 5 and 3 kDa). Antioxidant activities studied included reducing power, ferrous ion chelation, DPPH (1, 1- diphenyl-2- picrylhydrazyl) radical scavenging, and superoxide anion scavenging. Functional properties studied included emulsifying and foaming properties as well as fat and water binding capacity. Results showed significant differences (p
Extraction of gelatin using traditional acid-base pretreatment method has several limitations
such as time consuming and causes serious water pollution. Chemical treatment often being used
as an alternative process to overcome the weaknesses of the conventional method. However,
excessive chemical elements would damage the structure of the gelatin due to its high sensitivity
to the acid content. High Pressure Processing (HPP) is a novel and environmental friendly
method that has been suggested to assist gelatin extraction. Pressurization during pretreatment
could reduce the extraction time and amount of acid used. It also has a potential in enhancing the
properties of the gelatin extract and increasing the gelatin yield. In this research, One-Factor-
at-Time (OFAT) and optimization study were done to determine the optimum parameters for
extraction of gelatin assisted by HPP from red tilapia skin. Four parameters; applied pressure,
pressure holding time, ratio of acid to skin and extraction time have been selected for the OFAT
design and concentration of the gelatin extract and percentage of yield gelatin were evaluated.
From OFAT, optimum technical parameters for response surface optimization design were 250
MPa pressure, 7.5 ml of acid to 1 g of skin and 12 hours extraction time. Pressure holding
time was fixed for 10 min. FCCCD has been used for optimization study. Results from the
data shows that the optimum conditions for gelatin extraction from red tilapia skin were 250
MPa for pressure, 10 min of pressure holding time, 7.5 ml of acid for 1 g of skin and 12 hours
of extraction time while the maximum concentration and yield were 19.51 mg/ml and 32.04%
(320.4 mg/g), respectively. These findings proved that HPP could increase the concentration
and the yield of the gelatin while reducing the chemical waste and shortening the extraction
process.
In this research, a novel method was performed to obtain hydrogel with superior thermal stability by incorporation
of cellulose nanocrystals (CNC) into gelatin based hydrogel. Glutaraldehyde was used as cross-linker due to its high
chemical reactivity towards NH2
group on gelatin. Different ratio of gelatin/CNC hydrogel was produced in order to study
the effects of CNC towards the swelling behaviour and thermal stability of gelatin based hydrogel. The obtained hydrogel
was subjected to Fourier transform infrared (FTIR) to verify that gelatin had been cross-linked, swelling test with different
pH for swelling behaviour and thermogravimetric analysis (TGA) for thermal stability. The presence of C=N stretching
group in the FTIR spectrum for gelatin/CNC hydrogel indicated that the cross-linking reaction between gelatin monomer
had been successfully carried out. The hydrogel showed impressive pH sensitivity and maximum swelling was obtained
at pH3. The TGA results clearly showed that the incorporation of CNC into gelatin was able to produce hydrogel with
higher thermal stability compare to neat gelatin.
A 9-week study was conducted to compare dietary corn starch (CS) or tapioca starch (TS), with or without being pre-gelatinized (PG), on the growth, feeding efficiencies, plasma and muscle biochemistry, intestinal short chain fatty acids (SCFA), and liver glycogen of triplicate groups of 20 red hybrid tilapia (Orecohromis sp.). Various pellet characteristics were evaluated, along with their surface and cross sectional microstructure. The PG diets had significantly higher water stability, bulk density, and protein solubility, along with a smoother surface. Tilapia fed the TS diet had lower growth than had all other tilapia, but were significantly improved when diet was pre-gelatinized. In the PG treatments, intestinal SCFA significantly decreased while plasma glucose, cholesterol and triglycerides, as well as liver glycogen, significantly increased. Fish fed the CS diet had significantly more long chain polyunsaturated fatty acid than had those fed by other treatments. Pre-gelatinization may improve fish productivity and offer greater flexibility during aquafeed production.
Microencapsulation of water-soluble drugs using coacervation-phase separation method is very challenging, as these drugs partitioned into the aqueous polymeric solution, resulting in poor drug entrapment. For evaluating the effect of ovalbumin on the microencapsulation of drugs with different solubility, pseudoephedrine HCl, verapamil HCl, propranolol HCl, paracetamol, and curcuminoid were used. In addition, drug mixtures comprising of paracetamol and pseudoephedrine HCl were also studied. The morphology, encapsulation efficiency, particle size, and in vitro release profile were investigated. The results showed that the solubility of the drug determined the ratio of ovalbumin to be used for successful microencapsulation. The optimum ratios of drug, ovalbumin, and gelatin for water-soluble (pseudoephedrine HCl, verapamil HCl, and propranolol HCl), sparingly water-soluble (paracetamol), and water-insoluble (curcuminoid) drugs were found to be 1:1:2, 2:3:5, and 1:3:4. As for the drug mixture, the optimum ratio of drug, ovalbumin, and gelatin was 2:3:5. Encapsulated particles prepared at the optimum ratios showed high yield, drug loading, entrapment efficiency, and sustained release profiles. The solubility of drug affected the particle size of the encapsulated particle. Highly soluble drugs resulted in smaller particle size. In conclusion, addition of ovalbumin circumvented the partitioning effect, leading to the successful microencapsulation of water-soluble drugs.
Skin is a major by-product of the fisheries and aquaculture industries and is a valuable source of gelatin. This study examined the effect of triploidization on gelatin yield and proximate composition of the skin of African catfish (Clarias gariepinus). We further investigated the effects of two commonly used pesticides, chlorpyrifos (CPF) and butachlor (BUC), on the skin gelatin yield and amino acid composition in juvenile full-sibling diploid and triploid African catfish. In two separate experiments, diploid and triploid C. gariepinus were exposed for 21 days to graded CPF [mean measured: 10, 16, or 31 μg/L] or BUC concentrations [Mean measured: 22, 44, or 60 μg/L]. No differences in skin gelatin yield, amino acid or proximate compositions were observed between diploid and triploid control groups. None of the pesticide treatments affected the measured parameters in diploid fish. In triploids, however, gelatin yield was affected by CPF treatments while amino acid composition remained unchanged. Butachlor treatments did not alter any of the measured variables in triploid fish. To our knowledge, this study is the first to investigate changes in the skin gelatin yield and amino acid composition in any animal as a response to polyploidization and/or contaminant exposure.
Gelatin is widely used in pharmaceuticals as a protective coating, such as soft and hard capsule shells. However, the animal source of gelatin is a sensitive issue because certain gelatins such as porcine and bovine gelatins are not welcome in Halal, Kosher and Hindus' consumer goods. Recently, we have documented DNA barcoding and multiplex PCR platforms for discriminating porcine, bovine and fish gelatins in various fish and confectionary products; but those assays were not self-authenticating and also not tested in highly refined pharmaceutical products. To address this knowledge gap, here we report a self-authenticating multiplex PCR-restriction fragment length polymorphism (RFLP) assay to identify animal sources of various gelatin in pharmaceutical capsules. Three different restriction enzymes, BsaAI, Hpy188I and BcoDI were used to yield distinctive RFLP patterns for gelatin-based bovine (26, 94 bp), fish (97, 198 bp) and porcine (17, 70 bp) DNA in control experiments. The specificity was cross-tested against 16 non-target species and the optimised assay was used to screen gelatin sources in 30 halal-branded pharmaceuticals capsule shells. Bovine and porcine DNA was found in 27 and 3 of the 30 different capsules products. The assay was suitable for detecting 0.1 to 0.01 ng total DNA extracted from pure and mixed gelatins. The study might be useful to authenticate and monitor halal, kosher, vegetarian and Hindu compliant pharmaceuticals, foods and cosmetics.
Edible duck feet gelatin (DFG)-based biocomposites with different glycerol (GLY) contents (15%, 25%, and 35% of dried DFG) were prepared. Physicochemical, mechanical, barrier, and heat seal properties of DFG films were characterized and compared as an alternative to bovine gelatin film. Increasing glycerol from 15 to 35% decreased the TS and YM and EB and HS increased, in value of 42.54-7.27 and 1240-157.10MPa and 22.82-50.33% and 42.06-347.15N/m respectively. The water vapor permeability (WVP) and oxygen permeability (OP) of films were increased from 4.78 to 5.6×10-11gm-1Pa-1s-1 and from 3.97 to 33.99cm3mμ/m2 d kPa respectively. GAB model estimations showed monolayer water content of films increased with the increase of plasticizer content. Moisture sorption isotherm modelling exhibited a type II BET classification. Fourier transform infrared (FTIR) spectra showed shifted peak at approximately 1024cm-1, which was related to glycerol. The results show that the properties of DFG film are suitable for use as an alternative material to bovine gelatin film.
An orally-administered system for targeted, on-demand drug delivery to the gastrointestinal (GI) tract is highly desirable due to the high instances of diseases of that organ system and harsh mechanical and physical conditions any such system has to endure. To that end, we present an iron oxide nanoparticle/wax composite capsule coating using magnetic hyperthermia as a release trigger. The coating is synthesised using a simple dip-coating process from pharmaceutically approved materials using a gelatin drug capsule as a template. We show that the coating is impervious to chemical conditions within the GI tract and is completely melted within two minutes when exposed to an RF magnetic field under biologically-relevant conditions. The overall simplicity of action, durability and non-toxic and inexpensive nature of our system demonstrated herein are key for successful drug delivery systems.
Nasal injury following nasal surgery is an adverse consequence, and prompt treatment should be initiated. Nasal packing, either non-absorbable or absorbable, are commonly used after nasal surgery to prevent bleeding and promote wound healing. In the current study, a novel gelatine sponge crosslinked with genipin was evaluated for suitability to be used as nasal packing and compared to one of the frequently used commercial nasal packing made up of polyurethane. Gelatine at 7% and 10% (w/v) concentration were crosslinked with varying concentrations of genipin, 0.5%, 0.25%, and 0.2% (v/v). The gelatine sponges were further characterised by its water uptake ability, biodegradation, water vapour transmission rate, porosity, contact angle, chemical composition, crosslinking degree, and mechanical properties. The gelatine sponges absorbed five times more water than their dry weight and were degraded within five days. The water vapour transmission rate of the gelatine sponges was 1187.7 ± 430.2 g/(m-2 day) for 7% gelatine and 779.4 ± 375.5 g/(m-2 day) for 10% gelatine. Crosslinking of gelatine with genipin resulted in lower porosity and did not affect the wettability of gelatine sponge (contact angle: 95.3 ± 12.1° for 7% gelatine and 88.4 ± 7.2° for 10% gelatine). In terms of biodegradability, the gelatine sponges took 24-48 h to degrade completely. Genipin crosslinking improved the degradation resistance and mechanical strength of gelatine sponge. The physical and chemical properties of the gelatine sponge, i.e. biodegradability and mechanical durability, support its potential as nasal packing.
Chicken skin gelatin hydrolysates and peptides with angiotensin converting enzyme inhibitory (ACEI) activity were produced enzymatically using alcalase, pronase E, and collagenase before fractionation into
Introduction: Collagen and gelatin are essential protein in vertebrates and extensively used in various industries. Methods: In this study, acid-solubilized collagen and gelatin were extracted from the scales of three different species of freshwater fish namely Kelah (Tombroides), Tilapia (Oreochromis niloticus) and Snakehead fish (Channidae) and then further quantified using Bradford assay and separated by molecular weight using SDS-PAGE. Results: The extracted collagen in Tilapia fish scale was found to be the highest with 0.018 of protein absorbance among the other three fish; Kelah fish (0.017) and Snakehead fish (0.011). For gelatin, Snakehead fish scales showed the highest amount of total protein concentration followed by Tilapia and Kelah fish with 0.467, 0.144 and 0.037 μg/μL per g, respectively. Based on the SDS-PAGE results, collagen from all the three freshwater fishes were identified as a type 1 (molecular weight approximately from 95 to 130 kDa) collagen. As for gelatin, only gelatin from Snakehead fish scale was identified to be a type 1(molecular weight approximately from 95 to 130 kDa) while the other two freshwater fishes showed no clear band due to high viscosity of the gelatin produced. Conclusion: It can be said that the fishes investigated in this study have a potential to be the alternative source of collagen and gelatin.