Untargeted metabolomics using liquid chromatography-high resolution mass spectrometry and chemometrics for analysis of non-halal meats adulteration in beef meat.

OBJECTIVE: The adulteration of raw beef (BMr) with dog meat (DMr) and pork (PMr) becomes a serious problem because it is associated with halal status, quality, and safety of meats. This research aimed to develop an effective authentication method to detect non-halal meats (dog meat and pork) in beef using metabolomics approach. METHODS: Liquid chromatography-high resolution mass spectrometry (LC-HRMS) using untargeted approach combined with chemometrics was applied for analysis non-halal meats in BMr. RESULTS: The untargeted metabolomics approach successfully identified various metabolites in BMr DMr, PMr, and their mixtures. The discrimination and classification between authentic BMr and those adulterated with DMr and PMr were successfully determined using partial least square-discriminant analysis (PLS-DA) with high accuracy. All BMr samples containing non-halal meats could be differentiated from authentic BMr. A number of discriminating metabolites with potential as biomarkers to discriminate BMr in the mixtures with DMr and PMr could be identified from the analysis of variable importance for projection value. Partial least square (PLS) and orthogonal PLS (OPLS) regression using discriminating metabolites showed high accuracy (R2>0.990) and high precision (both RMSEC and RMSEE <5%) in predicting the concentration of DMr and PMr present in beef indicating that the discriminating metabolites were good predictors. The developed untargeted LC-HRMS metabolomics and chemometrics successfully identified non-halal meats adulteration (DMr and PMr) in beef with high sensitivity up to 0.1% (w/w). CONCLUSION: A combination of LC-HRMS untargeted metabolomic and chemometrics promises to be an effective analytical technique for halal authenticity testing of meats. This method could be further standardized and proposed as a method for halal authentication of meats.

Analysis of dog meat adulteration in beef meatballs using non-targeted UHPLC-Orbitrap HRMS metabolomics and chemometrics for halal authentication study.

Due to the different price and high quality, halal meat such as beef can be adulterated with non-halal meat with low price to get an economical price. The objective of this research was to develop an analytical method for halal authentication testing of beef meatballs (BM) from dog meat (DM) using a non-targeted metabolomics approach employing liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and chemometrics. The differentiation of authentic BM from that adulterated with DM was successfully performed using partial least square-discriminant analysis (PLS-DA) with high accuracy (R2X = 0.980, and R2Y = 0.980) and good predictivity (Q2 = 0.517). In addition, partial least square (PLS) and orthogonal PLS (OPLS) were successfully used to predict the DM added (% w/w) in BM with high accuracy (R2 > 0.990). A number of metabolites, potential for biomarker candidates, were identified to differentiate BM and that adulterated with DM. It showed that the combination of a non-targeted LC-HRMS Orbitrap metabolomics and chemometrics could detect up to 0.1% w/w of DM adulteration. The developed method was successfully applied for analysis of commercial meatball samples (n = 28). Moreover, pathway analysis revealed that beta-alanine, histidine, and ether lipid metabolism were significantly affected by dog meat adulteration. In summary, this developed method has great potential to be developed and used as an alternative method for analysis of non-halal meats in halal meat products.

Analysis of Pork in Beef Sausages Using LC-Orbitrap HRMS Untargeted Metabolomics Combined with Chemometrics for Halal Authentication Study.

Beef sausage (BS) is one of the most favored meat products due to its nutrition and good taste. However, for economic purposes, BS is often adulterated with pork by unethical players. Pork consumption is strictly prohibited for religions including Islam and Judaism. Therefore, advanced detection methods are highly required to warrant the halal authenticity of BS. This research aimed to develop a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) method to determine the halal authenticity of BS using an untargeted metabolomics approach. LC-HRMS was capable of detecting various metabolites in BS and BS containing pork. The presence of pork in BS could be differentiated using principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) with high accuracy. PLS-DA perfectly classified authentic BS and BS containing pork in all concentration levels of pork with R2X = (0.821), R2Y(= 0.984), and Q2 = (0.795). The level of pork in BS was successfully predicted through partial least squares (PLS) and orthogonal PLS (OPLS) chemometrics. Both models gave high R2 (>0.99) actual and predicted values as well as few errors, indicating good accuracy and precision. Identification of discriminating metabolites' potential as biomarker candidates through variable importance for projections (VIP) value revealed metabolites of 2-arachidonyl-sn-glycero-3-phosphoethanolamine, 3-hydroxyoctanoylcarnitine, 8Z,11Z,14Z-eicosatrienoic acid, D-(+)-galactose, oleamide, 3-hydroxyhexadecanoylcarnitine, arachidonic acid, and α-eleostearic acid as good indicators to detect pork. It can be concluded that LC-HRMS metabolomics combined with PCA, PLS-DA, PLS, and OPLS was successfully used to detect pork adulteration in beef sausages. The results imply that LC-HRMS untargeted metabolomics in combination with chemometrics is a promising alternative as an analytical technique to detect pork in sausage products. Further analysis of larger samples is required to warrant the reproducibility.

Application of Chromatographic and Spectroscopic-Based Methods for Analysis of Omega-3 (ω-3 FAs) and Omega-6 (ω-6 FAs) Fatty Acids in Marine Natural Products.

Omega-3 fatty acids v(ω-3 FAs) such as EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) and omega-6 fatty acids (ω-6 FAs) such as linoleic acid and arachidonic acid are important fatty acids responsible for positive effects on human health. The main sources of ω-3 FAs and ω-6 FAs are marine-based products, especially fish oils. Some food, supplements, and pharmaceutical products would include fish oils as a source of ω-3 FAs and ω-6 FAs; therefore, the quality assurance of these products is highly required. Some analytical methods mainly based on spectroscopic and chromatographic techniques have been reported. Molecular spectroscopy such as Infrared and Raman parallel to chemometrics has been successfully applied for quantitative analysis of individual and total ω-3 FAs and ω-6 FAs. This spectroscopic technique is typically applied as the alternative method to official methods applying chromatographic methods. Due to the capability to provide the separation of ω-3 FAs and ω-6 FAs from other components in the products, gas and liquid chromatography along with sophisticated detectors such as mass spectrometers are ideal analytical methods offering sensitive and specific results that are suitable for routine quality control.

Detection of Pork in Beef Meatballs Using LC-HRMS Based Untargeted Metabolomics and Chemometrics for Halal Authentication.

Adulteration of high-quality meat products using lower-priced meats, such as pork, is a crucial issue that could harm consumers. The consumption of pork is strictly forbidden in certain religions, such as Islam and Judaism. Therefore, the objective of this research was to develop untargeted metabolomics using liquid chromatography-high resolution mass spectrometry (LC-HRMS) combined with chemometrics for analysis of pork in beef meatballs for halal authentication. We investigated the use of non-targeted LC-HRMS as a method to detect such food adulteration. As a proof of concept using six technical replicates of pooled samples from beef and pork meat, we could show that metabolomics using LC-HRMS could be used for high-throughput screening of metabolites in meatballs made from beef and pork. Chemometrics of principal component analysis (PCA) was successfully used to differentiate beef meatballs and pork meatball samples. Partial least square-discriminant analysis (PLS-DA) clearly discriminated between halal and non-halal beef meatball samples with 100% accuracy. Orthogonal projection to latent structures-discriminant analysis (OPLS-DA) perfectly discriminated and classified meatballs made from beef, pork, and a mixture of beef-pork with a good level of fitness (R2X = 0.88, R2Y = 0.71) and good predictivity (Q2 = 0.55). Partial least square (PLS) and orthogonal PLS (OPLS) were successfully applied to predict the concentration of pork present in beef meatballs with high accuracy (R2 = 0.99) and high precision. Thirty-five potential metabolite markers were identified through VIP (variable important for projections) analysis. Metabolites of 1-(1Z-hexadecenyl)-sn-glycero-3-phosphocholine, acetyl-l-carnitine, dl-carnitine, anserine, hypoxanthine, linoleic acid, and prolylleucine had important roles for predicting pork in beef meatballs through S-line plot analysis. It can be concluded that a combination of untargeted metabolomics using LC-HRMS and chemometrics is promising to be developed as a standard analytical method for halal authentication of highly processed meat products.

Magnetic molecularly imprinted polymer nanoparticles for the extraction and clean-up of thiamethoxam and thiacloprid in light and dark honey.

In this work, new selective and sensitive dual-template molecularly imprinted polymer nanoparticles (MIPs) were synthesized and characterized. Sorbent MIPs were investigated for simultaneous extraction and clean-up of thiamethoxam and thiacloprid from light and dark honey samples. In this study, ultra-high-performance liquid chromatography-tandem mass spectrometry triple-quadrupole (UHPLC-MS/MS) (QQQ) was used to detect and quantify the pesticides. The kinetic model with adsorption kinetics of sorbent was investigated. The optimal adsorption conditions were 80 mg of polymer MIPs, a 30-min extraction time, and a pH of 7. The detection limit (LOD) and the quantification limit (LOQ) varied from 0.045 to 0.070 µg kg-1 and from 0.07 to 0.10 µg kg-1, respectively. The intra-day and inter-day precision (RSD, %) ranged from 1.3 to 2.0% and from 8.2 to 12.0%, respectively. The recovery of thiamethoxam and thiacloprid ranged from 96.8 to 106.5% and 95.3 to 104.4%, respectively, in light and dark honey samples.

Modified APHA closed-tube reflux colorimetric method for TOC determination in water and wastewater.

The analysis of total organic carbon (TOC) by the American Public Health Association (APHA) closed-tube reflux colorimetric method requires potassium dichromate (K2Cr2O7), silver sulfate (AgSO4), and mercury (HgSO4) sulfate in addition to large volumes of both reagents and samples. The method relies on the release of oxygen from dichromate on heating which is consumed by carbon associated with organic compounds. The method risks environmental pollution by discharging large amounts of chromium (VI) and silver and mercury sulfates. The present method used potassium monochromate (K2CrO4) to generate the K2Cr2O7 on demand in the first phase. In addition, miniaturizing the procedure to semi microanalysis decreased the consumption of reagents and samples. In the second phase, mercury sulfate was eliminated as part of the digestion mixture through the introduction of sodium bismuthate (NaBiO3) for the removal of chlorides from the sample. The modified method, the potassium monochromate closed-tube colorimetry with sodium bismuthate chloride removal (KMCC-Bi), generates the potassium dichromate on demand and eliminates mercury sulfate. The semi microanalysis procedure leads to a 60% reduction in sample volume and ≈ 33.33 and 60% reduction in monochromate and silver sulfate consumption respectively. The LOD and LOQ were 10.17 and 33.90 mg L-1 for APHA, and 4.95 and 16.95 mg L-1 for KMCC-Bi. Recovery was between 83 to 98% APHA and 92 to 104% KMCC-Bi, while the RSD (%) ranged between 0.8 to 5.0% APHA and 0.00 to 0.62% KMCC-Bi. The method was applied for the UV-Vis spectrometry determination of COD in water and wastewater. Statistics was done by MINITAB 17 or MS Excel 2016. ᅟ Graphical abstract.

A simple method for chromium speciation analysis in contaminated water using APDC and a pre-heated glass tube followed by HPLC-PDA.

In this study, a simple sample preparation method was developed for the determination of tri-and hexavalent chromium in water samples. It utilizes a pre-heated customized glass tube (CGT), to supply the heat energy required for the reaction of Cr(III) with ammonium pyrrolidinedithiocarbamate (APDC). The products of the Cr complexes, tris(1-pyrrolidinecarbodithioato)chromium(III) and bis(1-pyrrolidinecarbodithioato)[1-pyrrolidinecarbodithio(thioperoxoato)]chromium(III) were chromatographed with Shimadzu LC-20AT and Zobax Eclipse C18 (150mm × 4.6mm, 5µm) column using ACN: Water, (7:3, v/v) as the mobile phase. The concentration of Cr(III) ranged from 0.06mgL-1 to 0.09mgL-1 and that of Cr(VI) was between 0.02mgL-1 to 0.04mgL-1 in the samples. Percentage recoveries from spiked real samples were between 87% (tap water) to 110% (wastewater) for Cr(III) and 92% (pond water) to 117% (tap water) for Cr(VI). The limits of detection (LODs) were 0.0029mgL-1 and 0.0014mg/L-1 for Cr(III) Cr(VI) respectively. While the limits of quantitation (LOQs), were 0.0098mgL-1 and 0.0047mgL-1 for Cr(III) and Cr(VI) respectively. Method precision (RSD (%)) was 3.3% and 3.5% for Cr(III) and Cr(VI) respectively. The developed method was applied for the speciation analysis of chromium in drinking water, tap water, wastewater, river water, and pond water samples. Our findings proved the method is simple and inexpensive. The method was validated by the analysis of a certified reference material (CRM) SLRS-4. The percentage recovery and RSD(%) from the spiked CRM were 91% and 115% and 0.32% and 1.4% for Cr(III) and Cr(VI) respectively.

Geochemical characteristics of rare earth elements in different types of soil: A chemometric approach.

Rare earth elements (REEs) are becoming significant due to their huge applications in many industries, large-scale mining and refining activities. Increasing usage of such metals pose negative environmental impacts. In this research ICP-MS has been used to analyze soil samples collected from former ex-mining areas in the depths of 0-20 cm, 21-40 cm, and 41-60 cm of residential, mining, natural, and industrial areas of Perak. Principal component analysis (PCA) revealed that soil samples taken from different mining, industrial, residential, and natural areas are separated into four clusters. It was observed that REEs were abundant in most of the samples from mining areas. Concentration of the rare elements decrease in general as we move from surface soil to deeper soils.

Chemical speciation and bioavailability of rare earth elements (REEs) in the ecosystem: a review.

Rare earths (RE), chemically uniform group of elements due to similar physicochemical behavior, are termed as lanthanides. Natural occurrence depends on the geological circumstances and has been of long interest for geologist as tools for further scientific research into the region of ores, rocks, and oceanic water. The review paper mainly focuses to provide scientific literature about rare earth elements (REEs) with potential environmental and health effects in understanding the research. This is the initial review of RE speciation and bioavailability with current initiative toward development needs and research perceptive. In this paper, we have also discussed mineralogy, extraction, geochemistry, analytical methods of rare earth elements. In this study, REEs with their transformation and vertical distribution in different environments such as fresh and seawater, sediments, soil, weathering, transport, and solubility have been reported with most recent literature along key methods of findings. Speciation and bioavailability have been discussed in detail with special emphasis on soil, plant, and aquatic ecosystems and their impacts on the environment. This review shows that REE gained more importance in last few years due to their detrimental effects on living organisms, so their speciation, bioavailability, and composition are much more important to evaluate their health risks and are discussed thoroughly as well.

Assessing anthropogenic levels, speciation, and potential mobility of rare earth elements (REEs) in ex-tin mining area.

A study was carried out to determine the level of rare earth elements (REEs) in water and sediment samples from ex-mining lakes and River in Kinta Valley, Perak, Malaysia. Surface water and sediments from an ex-mining lake and Kinta River water samples were analyzed for REEs by inductively coupled plasma mass spectrometry. The total concentration of REEs in the ex-mining lake water samples and sediments were found to be 3685 mg/l and 14159 mg/kg, respectively, while the total concentration of REEs in Kinta River water sample was found to be 1224 mg/l. REEs in mining lake water were found to be within 2.42 mg/l (Tb) to 46.50 mg/l (Ce), while for the Kinta River, it was 1.33 mg/l (Ho) to 29.95 mg/l (Ce). Sediment samples were also found with REEs from 9.81 mg/kg (Ho) to 765.84 mg/kg (Ce). Ce showed the highest average concentrations for mining lake (3.88 to 49.08 mg/l) and Kinta River (4.44 to 33.15 mg/l) water samples, while the concentration of La was the highest (11.59 to 771.61 mg/kg) in the mining lake sediment. Lu was shown to have the highest enrichment of REEs in ex-mining lake sediments (107.3). Multivariate statistical analyses such as factor analysis and principal component analysis indicated that REEs were associated and controlled by mixed origin, with similar contributions from anthropogenic and geogenic sources. The speciation study of REEs in ex-tin mining sediments using a modified five-stage sequential extraction procedure indicated that yttrium (Y), gadolinium (Gd), and lanthanum (La) were obtained at higher percentages from the adsorbed/exchanged/carbonate fraction. The average potential mobility of the REEs was arranged in a descending order: Yb > Gd > Y = Dy > Pr > Er > Tm > Eu > Nd > Tb > Sc > Lu > Ce > La, implying that under favorable conditions, these REEs could be released and subsequently pollute the environment.

Release, deposition and elimination of radiocesium ((137)Cs) in the terrestrial environment.

Radionuclide contamination in terrestrial ecosystems has reached a dangerous level. The major artificial radionuclide present in the environment is (137)Cs, which is released as a result of weapon production related to atomic projects, accidental explosions of nuclear power plants and other sources, such as reactors, evaporation ponds, liquid storage tanks, and burial grounds. The release of potentially hazardous radionuclides (radiocesium) in recent years has provided the opportunity to conduct multidisciplinary studies on their fate and transport. Radiocesium's high fission yield and ease of detection made it a prime candidate for early radio-ecological investigations. The facility setting provides a diverse background for the improved understanding of various factors that contribute toward the fate and transfer of radionuclides in the terrestrial ecosystem. In this review, we summarize the significant environmental radiocesium transfer factors to determine the damaging effects of radiocesium on terrestrial ecosystem. It has been found that (137)Cs can trace the transport of other radionuclides that have a high affinity for binding to soil particles (silts and clays). Possible remedial methods are also discussed for contaminated terrestrial systems. This review will serve as a guideline for future studies of the fate and transport of (137)Cs in terrestrial environments in the wake of the Fukushima Nuclear Power Plant disaster in 2011.

A Comparative Study of Selected Trace Element Content in Malay and Chinese Traditional Herbal Medicine (THM) Using an Inductively Coupled Plasma-Mass Spectrometer (ICP-MS).

A total of 60 products of traditional herbal medicine (THM) in various dosage forms of herbal preparation were analyzed to determine selected trace elements (i.e., Zn, Mn, Cu, Cd, and Se) using ICP-MS. Thirty types of both Chinese and Malay THMs were chosen to represent each population. The closed vessel acid microwave digestion method, using CEM MARS 5, was employed for the extraction of the selected trace elements. The digestion method applied was validated by using certified reference material from the Trace Element in Spinach Leaves (SRM1570a). The recoveries of all elements were found to be in the range of 85.3%-98.9%. The results indicated that Zn, Mn, Cu, Cd and Se have their own trends of concentrations in all samples studied. The daily intake concentrations of the elements were in the following order: Mn > Zn > Cu > Se > Cd. Concentrations of all five elements were found to be dominant in Chinese THMs. The essentiality of the selected trace elements was also assessed, based on the recommended daily allowance (RDA), adequate intake (AI) and the United States Pharmacopeia (USP) for trace elements as reference. The concentrations of all elements studied were below the RDA, AI and USP values, which fall within the essential concentration range, except for cadmium.