16 research outputs found
Development and validation of amino acid analysis methods in gelatin and gelatin-based products using high-performance liquid chromatography
Get PDFThe similarity of physicochemical properties among gelatin especially the porcine and
bovine gelatin has sparked skepticism among Muslim consumers towards gelatinbased
commercial products. This study was aimed to develop and validate a reversephase
high performance liquid chromatography (RP-HPLC) method of amino acid
analysis in gelatin and differentiate the bovine, porcine and fish gelatin as an
ingredient and in gelatin-based commercial products using the principal component
analysis (PCA). The analytical method used was amino acid analysis that using 6-
aminoquinolyl-N-hydroxysuccinimidyl carbamate as derivatization reagent and the
chromatographic separation was determined by RP-HPLC coupled with a
fluorescence detector. Method development was conducted according to ISO 17025
guidelines. In-house method validation revealed that the method was selectively
performing a good chromatographic separation for 18 amino acids; the detection and
quantitation limit were ranged from 5.68–12.62 and 36.0–39.0 pmol/μl, respectively;
no matrix effect was observed and the linearity range was 37.5–1000 pmol/μl. Method
precision revealed by HorRat values was significantly less than 2 and the method
recoveries had a range of 80–115%. The uncertainty evaluation was estimated on the
basis of the method validation data. The uncertainty of method precision, μ(P), method
recovery, μ(R) and measurement of standard, μ(Std) for overall amino acids are within
a concentration range of 0.024 to 0.113 pmol/μl, 0.006 to 0.10 pmol/μl and 0.010 to
0.0297 pmol/μl, respectively. PCA has assisted the process of distinguishing the
bovine, porcine and fish gelatin. Data pre-treatments such as centering and area
normalization were performed to reduce the variances of variables in dataset. Database
for three gelatins were established through this work and were verified by samples
from gelatin-based commercial products. Data analysis demonstrated that the fish
gelatin was correlated to threonine, serine and methionine on the positive side of
principal component (PC) 1; bovine gelatin was correlated to the non-polar side chains
amino acids that were proline, hydroxyproline, leucine, isoleucine and valine on the
negative side of PC1 and porcine gelatin was correlated to the polar side chains amino
acids that were aspartate, glutamic acid, lysine and tyrosine on the negative side of PC2. The lowest detection value for adulteration of porcine gelatin in bovine gelatin
was determined at 0.05% (w/w) of porcine gelatin. The extraction of gelatin from
gelatin-based commercial products was successful by samples clean-up using acetone
solvent and modification on several parameters of amino acid analysis method
specifically the sample digestion process. Gelatin used in the commercial products
was verified using gelatin database and the results revealed that products made of
porcine gelatin can be differentiated from the products that were made of bovine
gelatin. This quantitative method was very useful as an alternative method for halal
products authentication via laboratory testing
Use of liquid chromatographic methods for gelatin differentiation in halal verification
No full textA wide diversity of gelatin sources that supported by the largest scale of porcine gelatin production in a global market has raised doubts among Muslim consumers towards gelatin-based ingredients products. The skepticism has been addressed through the implementation of food labelling regulations that require manufacturers to state the source of gelatin on the packaging. Since the application of halal certification is voluntary and not compulsory for industry players, hence it gives significant drawbacks. Without strictly monitoring and enforcement, the probability of contamination with non-halal materials during the production process and lacking of halal integrity particularly in the supply chain of raw materials may increase the number of masbooh and non-compliance products according to the Islamic law. Therefore, a trust on the halal products must be strengthened with the scientific evidence. This paper attempts to critically review on liquid chomatographic methods in determining the species origin of gelatin. It covers series of technological changes from a simple of high performance liquid chromatography (HPLC) to the system that coupled with mass spectrometry. A large similarity in amino acids profile and polypeptide sequences of the mammalian gelatin will be the main challenge in developing the analytical methods. Matrix interference in the gelatin-based products, adulteration and fraud in the mixed ingredients are an inevitable, hence it need to be tackled and solved. These analytical methods are expected to be used as tools to provide scientific evidences for halal verification on the gelatin-based products
RP-HPLC method using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate incorporated with normalization technique in principal component analysis to differentiate the bovine, porcine and fish gelatins
No full textThe amino acid compositions of bovine, porcine and fish gelatin were determined by amino acid analysis using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate as derivatization reagent. Sixteen amino acids were identified with similar spectral chromatograms. Data pre-treatment via centering and transformation of data by normalization were performed to provide data that are more suitable for analysis and easier to be interpreted. Principal component analysis (PCA) transformed the original data matrix into a number of principal components (PCs). Three principal components (PCs) described 96.5% of the total variance, and 2 PCs (91%) explained the highest variances. The PCA model demonstrated the relationships among amino acids in the correlation loadings plot to the group of gelatins in the scores plot. Fish gelatin was correlated to threonine, serine and methionine on the positive side of PC1; bovine gelatin was correlated to the non-polar side chains amino acids that were proline, hydroxyproline, leucine, isoleucine and valine on the negative side of PC1 and porcine gelatin was correlated to the polar side chains amino acids that were aspartate, glutamic acid, lysine and tyrosine on the negative side of PC2. Verification on the database using 12 samples from commercial products gelatin-based had confirmed the grouping patterns and the variables correlations. Therefore, this quantitative method is very useful as a screening method to determine gelatin from various sources
Validation of a reverse-phase high-performance liquid chromatography method for the determination of amino acids in gelatins by application of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent
No full textIn-house method validation was conducted to determine amino acid composition in gelatin by a pre-column derivatization procedure with the 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent. The analytical parameters revealed that the validated method was capable of selectively performing a good chromatographic separation for 18 amino acids in less than 40 min; the overall detection and quantitation limit for amino acids fell into ranges of 5.68–12.48 and 36.0–39.0 pmol/μl, respectively; the matrix effect was not observed, and the linearity range was 37.5–1000 pmol/μl. The accuracy (precision and recovery) analyses of the method were conducted under repeatable conditions on different days in random order. Method precision revealed by HorRat values was significantly less than 2, except for histidine with a precision of 2.19, and the method recoveries had a range of 80–115% except for alanine which was recovered at 79.4%. The findings were reproducible and accurately defined, and the method was found to be suited to routine analysis of amino acid composition in gelatin-based ingredients
Usability evaluation of virtual umrah based on user centered design from expert user view
No full textWith the emergence of the learning materials volume, the developer has to face many challenges to develop an effective and efficient application to support the learning process. The objective of this study is to evaluate usability of Virtual Umrah application based on expert views using User-Centered Design approach. User-Centered Design is a multi-stages process that requires the designers to analyze and verify the requirement of a user by involving the user throughout the whole processes. The findings of the usability evaluations are for errors, efficiency and satisfaction level, the individual mean score is 4.00, 4.20 and 4.40. While for the memorability and learnability component, the mean score for the items are below than 4.00 which are 3.80 and 3.70 respectively. These findings demonstrate that the developed application is significantly needed by the pilgrims and individuals in order to assist them in preparation and performing Umrah
Incorporation of partial least squares-discriminant analysis with Ultra-High-Performance Liquid Chromatography Diode-Array Detector for authentication of skin gelatine sources
Get PDFThis research seeks to (1) authenticate sources of skin gelatine by combining putative 17 aminoacids (AAs) analysis with chemometrics by Ultra-High-Performance Liquid ChromatographyDiode-Array Detector (UHPLC-DAD) and (2) create AA profiles in skin gelatines. Theclassification capability of partial least square-discriminant analysis (PLS-DA) models wasassessed to determine the most effective discriminant model. Principal component analysis(PCA) with quartimax rotation was utilised to accurately organise gelatine clusters and assign thesignificantly contributing AAs to each cluster. The PLS-DA model with 13 AAs (PLS-DAVIPAA)outperformed the PLS-DA model with 17 AAs (PLS-DAAA) because its R2Y (0.938), R2X (0.881),and Q2 (0.929) values were greater. With 13 significant AAs, the PLS-DAVIPAA model obtainedcluster classification accuracy of 100% on training and cross-validation datasets and 93.3% ontesting and verification datasets. The chemical structure of gelatines may shed light on theinteractions between AAs. Following six quartimax rotations, the gelatines were groupedcorrectly. The PCA showed the dominant presence of these AAs: L-Valine, L-Phenylalanine andL-Tyrosine in porcine gelatine; Glycine, L-Threonine, L-Arginine, L-Methionine, L-Histidine andL-Serine in fish gelatine; and L-Hydroxyproline, L-Leucine and L-Proline in bovine gelatine. Theauthority could use this technique to set a standard for authenticating skin gelatine samples
Estimation of uncertainty from method validation data: application to a reverse-phase high-performance liquid chromatography method for the determination of amino acids in gelatin using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent
Get PDFA detailed procedure for estimating uncertainty according to the Laboratory of Government Chemists/Valid Analytical Measurement (LGC/VAM) protocol for determination of 18 amino acids in gelatin is proposed. The expanded uncertainty was estimated using mainly the method validation data (precision and trueness). Other sources of uncertainties were contributed by components in standard preparation measurements. The method scope covered a single matrix (gelatin) under a wide range of analyte concentrations. The uncertainty of method precision, μ(P) was 0.0237–0.1128 pmol μl−1 in which hydroxyproline and histidine represented the lowest and highest values of uncertainties, respectively. Proline and phenylalanine represented the lowest and highest uncertainties value for method recovery, μ(R) that was estimated within 0.0064–0.0995 pmol μl−1. The uncertainties from other sources, μ(Std) were 0.0325, 0.0428 and 0.0413 pmol μl−1 that were contributed by hydroxyproline, other amino acids and cystine, respectively. Hydroxyproline and phenylalanine represented the lowest and highest values of expanded uncertainty, U(y) that were determined at 0.0949 and 0.2473 pmol μl−1, respectively. The data were accurately defined and fulfill the technical requirements of ISO 17025:2005
Strategic approaches to halal lipid authentication using instrumental, chemometric, and traceability techniques
No full textThis book explores the complex field of halal lipids, combining contemporary scientific discoveries with traditional Islamic beliefs. This thorough handbook, which covers everything from lipid extraction and microbial lipid applications to cutting-edge applications in cosmetics and active packaging, is intended for researchers, business executives, legislators, and students. With an emphasis on sustainability and traceability, the chapters examine the scientific, technological, and economic facets of halal lipids. This book offers insights into how science, innovation, and Islamic values may come together to promote growth and consumer trust as the halal business adapts to meet demands from throughout the world
Bioinformatics tools assist in the screening of potential porcine-specific peptide biomarkers of gelatin and collagen for halal authentication
Get PDFGelatin and collagen are two animal-derived ingredients that are widely used in various industries. Both have distinctive physico-chemical characteristic that made them ingredients of interest for many industrial players to be applied as there are vast arrays of usage in the food, cosmetic and biomedical fields. However, the origin of gelatin and collagen poses ethical and religious concerns, especially for Muslims and Jews who have restrictions on food consumption. Porcine by-products are of concern for religious and health reasons, and there is a demand for precise and reliable detection techniques. The limitation of DNA detection is due to extreme environment in food processing which results in low extractability of DNA. Therefore, peptide-based detection using mass spectrometry is required. However, identify the suitable marker is like searching needle in haystacks. Hence, combination of bioinformatics and mass spectrometry is proposed. This study aims to identify the specific peptide biomarkers by employing bioinformatics technique which can be applied to identify gelatin and collagen sources with the aid of mass spectrometry. In these approach, combination of Petunia Trans-Proteomic Pipeline (TPP, version 5.2.0) and sequence alignment ClustalW were applied to facilitate the MS data (LC-QTOF-MS) and peptide identification. As a result, 69 fasta file of protein sequence from both UniProtKB and NCBInr have been collected, 81 collagen peptides sequence and 118 gelatine peptides has been attainable that have the potential to distinguish different species. In conclusion, in silico protein sequence approaches helps to enable rapid screening of proteotypic peptides that can serve as species biomarkers proficiently. © 2024 Malaysian Society of Applied Biology
Food forensics on gelatine source via ultra‑high‑performance liquid chromatography diode‑array detector and principal component analysis
Get PDFThis study provided a step-by-step procedure to investigate the distribution of 17 amino acids (AAs) in 50 fish, 50 bovine and 54 porcine gelatines using Ultra-High-Performance Liquid Chromatography Diode-Array Detector (UHPLC–DAD) with the incorporation of principal component analysis (PCA). Dataset pre-processing step, including outlier removal, analysis of variance (ANOVA), dataset adequacy test, dataset transformation and correlation test was performed before the PCA. The method rendered linearity range of 37.5–1000 pmol/µL and accuracy of 85–111% recovery. The bovine and porcine gelatines showed a similar ranking while the l-Alanine (Ala), l-Arginine (Arg) and l-Glutamic acid (Glu) concentrations had differed the fish gelatine from the bovine and porcine gelatines. The PCA, which explained 77.013% cumulative variability at eigenvalue of 5.436, showed AAs with strong FL in PC1 had polar and nonpolar side chains while AAs with strong FL in PC2 had polar side chain. The AAs with moderate and weak FL in PC1 had a nonpolar side chain. The AAs with strong FL of in PC1 were also the same AAs with 7, 6 and 5 strong CMs as determined in the correlation test. The second PCA showed that the l-Serine (Ser), Arg, Glycine (Gly), l-Threonine (Thr), l-Methionine (Met), l-Histidine (His) and L-Hydroxyproline (Hyp) were significant in fish gelatine; Hyp, Met, Thr, Ser, His, Gly, and Arg in bovine gelatine; and l-Proline (Pro), l-Tyrosine (Tyr), l-Valine (Val), l-Leucine (Leu), and l-Phenylalanine (Phe) in porcine gelatine. The 100% fish, bovine and porcine gelatines accommodated grouping 1, 2 and 3, respectively, which proved that AAs with strong FL (Hyp, His, Ser, Arg, Gly, Thr, Pro, Tyr, Met, Val, Leu and Phe) were the significant AAs and becomes the biomarkers to identify the gelatine source. From this study, the PCA was a useful tool to analyse a multivariate dataset that could provide an in-depth understanding of AA distributions as compared to ANOVA and correlation test
