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Moniliformin (MON) is a widespread emerging mycotoxin often occurring in maize at significant levels. Few published studies investigated MON redistribution in maize-derived products for human consumption; to better understand this issue, 5 maize lots with different levels of MON contamination were processed following an industrial milling process to evaluate the redistribution of the mycotoxin in final products (grits), by-products destined to feed (bran and flour) and cleaning waste. MON was quantified by LC-MS/MS after the purification step through the SPE column; moreover, a confirmatory method based on MON derivatization with 1,2-diamino-4,5-dichlorobenzene was developed. Relevant MON reduction was obtained after sieve cleaning, scourer process, and optical sorting, achieving a decrement of the concentration level close to 70%. The following other milling procedures showed a limited reduction from cleaned maize to small and large grits; considering the entire industrial process, the reduction percentage of MON contamination in the final products was 80.9 ± 9.3% and 81.0 ± 6.7% for small and large grits, respectively. The flaking process showed a very limited reduction of MON, close to 10%. Considering the widespread of MON occurrence in maize, the study highlights the importance of cleaning steps to achieve a low risk of exposure for the consumer., (© 2024. The Author(s).)

An infrared squaraine dye was utilized to detect Cu 2+ in solvents based on H-aggregates of squaraine dye. H-aggregates are a type of aggregation with enhanced photophysical properties compared to monomers. In the presence of a Ca 2+ solution, F-Cl offers exceptional H-aggregators that can be transformed into monomers by adding Cu 2+ . Furthermore, this mode successfully demonstrated fluorescence changes in HeLa cells cultured in vitro after the addition of Ca 2+ or Cu 2+ . A highly specific detection of Cu 2+ was achieved using this transformation mode., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Introduction: In the period between 1997 and 2010, sibutramine-containing drugs were widely prescribed for obesity and over-weight management. Due to safety concerns, in 2010 all medicines containing sibutramine were urgently withdrawn from the USA and European pharmaceutical market. Although sibutramine is no longer available in pharmaceutical products, there have been numerous reports of mislabeled weight-loss dietary supplements containing sibutramine., (This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Fusarium proliferatum and Fusarium subglutinans are common pathogens of maize which are known to produce mycotoxins, including moniliformin (MON) and fumonisins (FBs). Fungal secondary metabolism and response to oxidative stress are interlaced, where hydrogen peroxide (H 2 O 2 ) plays a pivotal role in the modulation of mycotoxin production. The objective of this study is to examine the effect of H 2 O 2 -induced oxidative stress on fungal growth, as well as MON and FBs production, in different isolates of these fungi. When these isolates were cultured in the presence of 1, 2, 5, and 10 mM H 2 O 2 , the fungal biomass of F. subglutinans isolates showed a strong sensitivity to increasing oxidative conditions (27-58% reduction), whereas F. proliferatum isolates were not affected or even slightly improved (45% increase). H 2 O 2 treatment at the lower concentration of 1 mM caused an almost total disappearance of MON and a strong reduction of FBs content in the two fungal species and isolates tested. The catalase activity, surveyed due to its crucial role as an H 2 O 2 scavenger, showed no significant changes at 1 mM H 2 O 2 treatment, thus indicating a lack of correlation with MON and FB changes. H 2 O 2 treatment was also able to reduce MON and FB content in certified maize material, and the same behavior was observed in the presence and absence of these fungi, highlighting a direct effect of H 2 O 2 on the stability of these mycotoxins. Taken together, these data provide insights into the role of H 2 O 2 which, when increased under stress conditions, could affect the vegetative response and mycotoxin production (and degradation) of these fungi.

Six Australian and five overseas complementary medicines (CM) and meal replacement shake products were analysed for potential adulteration with two common active pharmaceutical ingredients, caffeine and sibutramine, using thin-layer chromatography and mass spectrometry. The declared amount of caffeine in each product was also reviewed. Finally, the products were examined for heavy metal contamination using inductively coupled plasma-mass spectrometry. The results showed that there was no detected adulteration of either caffeine (for those products that did not list caffeine as an ingredient) or sibutramine in the 11 products; however, based on the product labels, one Australian and one overseas (two in total) CM product contained more than the maximum daily safety limit (400 mg) of caffeine. Potentially excessive lead and/or chromium was detected in six products, including four Australian products and two products purchased online. One Australian CM product appeared to contain these heavy metals at concentrations at, or exceeding, the safety limits specified in the United States Pharmacopeia or set by the World Health Organization. The overconsumption of caffeine and heavy metals has the potential of causing significant health effects in consumers.

With an increase in the obese population, the indiscriminate demand for anti-obesity drugs for rapid weight loss or maintenance has grown. As a result, illegal substances that could induce unexpected negative health effects or fatal side effects are being produced and mixed into consumer products. In the present study, the metabolites of five major illegal anti-obesity drugs are analyzed for the first time. Our data can be utilized to identify related compounds and predict their toxicological effects. Didesmethylsibutramine, desmethylsibutramine, homosibutramine, chlorosibutramine, and benzylsibutramine were metabolized in in vitro and in vivo models, and the metabolites were identified using liquid chromatography quadrupole-time of flight mass spectrometry (LC-Q-TOF-MS) and tandem mass spectrometry (LC-Q-TOF-MS/MS). The in vivo metabolite analysis was carried out using urine and feces samples from rats, and the in vitro metabolite analysis was performed by incubating the analogues with human liver microsomes. We found that each sibutramine analogue was metabolized into several constituents: 2 (M1-2), 5 (M1-5), 11 (M1-11), 7 (N1-7), and 5 (O1-5). In conclusion, our metabolic study could be used for toxicological detection of illegal obesity treatments and metabolite identification in forensic cases., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Immunochromatographic assay (ICA) has been used widely for the onsite monitoring of illegal additives due to its simplicity, speed, and low cost. However, a scanner is commonly required for ICA to achieve quantitative results. In this work, we developed a visual semi-quantitative ICA for sibutramine, a banned additive in diet foods, without the need for a scanner for measurement. Monoclonal antibodies specific for sibutramine were raised and conjugated with upconversion nanoparticles (UCNPs) as the luminescent tracer. ICA was developed by employing multiple test lines to achieve the semi-quantitative detection of sibutramine. Based on the optimal conditions, the cutoff levels (limit of quantitation, LOQ) of T1 line, T2 line, T3 line, and T4 line were 0.02 μg/mL, 0.15 μg/mL, 1.0 μg/mL, and 7.5 μg/mL, respectively, in buffer system. The ICA demonstrated a LOQ at 0.2 mg/kg for sibutramine in diet food samples. The assay (including pretreatment) can be finished within 30 min without the aid of other instruments, except a laser pen. No false positive or false negative results were observed. The results indicated that the proposed method was reliable, simple, and rapid for the screening of sibutramine abuse in diet food samples.

The food irradiation marker, 2-dodecylcyclobutanone (2-DCB), assayed by SPME provides a fast and simple method to estimate the irradiation history of fat-containing food products. The SPME conditions were optimized to maximize the extraction of 2-DCB from chicken jerky treats (CJT) irradiated at low (5 kGy) and high (50 kGy) doses. The extracted 2-DCB was measured using GC-MS in selected ion mode (m/z 98, and 112). Water dilution (1:5) was needed to mobilize 2-DCB and allow partition to the headspace form the CJT matrix. Increasing the incubation temperature to 80 °C resulted in higher response. Spiking control jerky samples with 2-DCB from 10 to 150 ng/g CJT compared with spiking water revealed a significant food matrix effect. This method provides a fast, simple, and environmental friendly alternative for the existing solvent extraction methods., (© 2020 Institute of Food Technologists®.)

The main objective of this study was to screen, for the first time, the natural occurrence of non-regulated fungal metabolites in 204 maize samples harvested in Serbia in maize growing seasons with extreme drought (2012), extreme precipitation and flood (2014) and moderate drought conditions (2013 and 2015). In total, 109 non-regulated fungal metabolites were detected in examined samples, whereby each sample was contaminated between 13 and 55 non-regulated fungal metabolites. Moniliformin and beauvericin occurred in all samples collected from each year. In samples from year 2012, oxaline, questiomycin A, cyclo (l-Pro-l-Val), cyclo (l-Pro-l-Tyr), bikaverin, kojic acid and 3-nitropropionic acid were the most predominant (98.0-100%). All samples from 2014 were contaminated with 7-hydroxypestalotin, 15-hydroxyculmorin, culmorin, butenolid and aurofusarin. Bikaverin and oxaline were quantified in 100% samples from 2013 and 2015, while 3-nitropropionic acid additionally occurred in 100% samples from 2015., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

As an important cellular signal transduction messenger, Ca 2+ has the capability to regulate cell function and control many biochemical processes, including metabolism, gene expression, and cell survival and death. Here, we introduce an accessible method for the photoactivation of Ca 2+ channels mediated by squaraine (SQ) to rapidly induce cellular Ca 2+ release and activate signal transduction. With a short preparation time, the maximum Ca 2+ concentration increase could reach approximately 450% in 30 s, resulting from marked Ca 2+ release channel opening in the endoplasmic reticulum (ER). This release was enhanced by another target location of SQ, that is, the outer mitochondrial-associated membrane where Ca 2+ channels accumulate, and by the consequent large amounts of reactive oxygen species resulting from the respiratory chain activity stimulated by Ca 2+ load. We used this method to investigate cellular signal transduction in different cancer cells and revealed rapid intracellular Ca 2+ flow, unidirectional intercellular signaling processes, and neuronal signaling activity, which demonstrated the potential and convenience of the method for routine Ca 2+ research.

New methods are described for the construction of targeted fluorescence probes for imaging cancer and the assessment of tumor targeting performance in a living mouse model. A novel noncovalent assembly process was used to fabricate a set of structurally related targeted fluorescent probes with modular differences in three critical assembly components: the emission wavelength of the squaraine fluorochrome, the number of cRGDfK peptide units that target the cancer cells, and the length of the polyethylene glycol chains as pharmacokinetic controllers. Selective targeting of cancer cells was proven by a series of cell microscopy experiments followed by in vivo imaging of subcutaneous tumors in living mice. The mouse imaging studies included a mock surgery that completely removed a fluorescently labeled tumor. Enhanced tumor accumulation due to probe targeting was first evaluated by conducting Single Agent Imaging (SAI) experiments that compared tumor imaging performance of a targeted probe and untargeted probe in separate mouse cohorts. Although there was imaging evidence for enhanced tumor accumulation of the targeted probe, there was moderate scatter in the data due to tumor-to-tumor variability of the vasculature structure and interstitial pressure. A subsequent Paired Agent Imaging (PAI) study coinjected a binary mixture of targeted probe (with emission at 690 nm) and untargeted probe (with emission at 830 nm) into the same tumor-burdened animal. The conclusion of the PAI experiment also indicated enhanced tumor accumulation of the targeted probe, but the statistical significance was much higher, even though the experiment required a much smaller cohort of mice. The imaging data from the PAI experiment was analyzed to determine the targeted probe's Binding Potential (BP) for available integrin receptors within the tumor tissue. In addition, pixelated maps of BP within each tumor indicated a heterogeneous spatial distribution of BP values. The results of this study show that the combination of fluorescent probe preassembly and PAI is a promising new way to rapidly develop targeted fluorescent probes for tumors with high BP and eventual use in clinical applications such as targeted therapy, image guided surgery, and personalized medicine.

Sibutramine is a highly effective drug for the treatment of obesity. In this regard, unscrupulous manufacturers can add sibutramine as a biologically active synthetic substance prohibited for use in the composition of dietary supplements. Thus, the problem associated with the illegal circulation of such dietary supplements is especially actual, given the scale of the sale of these products. The development and validation of methods for the determination of sibutramine in dietary supplements for slimming (anorexigenic action) for the purpose of quality control in order to ensure the quality of dietary supplements when introduced into civil circulation at customs and on the market. The aim of the study - to develop a method for the quantitative determination of sibutramin in dietary supplements for weight loss by high performance liquid chromatography (HPLC) and to validate it. Material and methods . For the quantitative determination of sibutramine in dietary supplements, we used an Agilent 1100 high performance liquid chromatograph with a UV-detector. The stationary phase was a chromatographic column C 18 NUCLEOSIL 4.6×150 mm, particle size 5 μm. The mobile phase contained 0.05 M formate buffer pH 4.0 and acetonitrile in a ratio of 40:60 (by volume). Results and discussion . A methodology has been developed for the determination of sibutramine in dietary supplements for weight loss, which makes it possible to control the quality of dietary supplements. Based on the obtained chromatograms, the specificity was determined; the plant components did not influence the determination of sibutramine in model mixtures. The suitability of the chromatographic system was determined: the retention factor of the compound - 2.222 (more than 2.0), N - 5776 theoretical plates (more than 5000), T peak of sibutramine - 0.939 (not more than 1.5). Within the analytical area of the linearity method: R 2 =0.9993 (over 0.9950). ε=0.46% (does not exceed 1.5%), the confidence interval includes the value of 100%, the calculated value of the Student's criterion tcalc (2.47) was less than the tabular ttable (2.80), which proved the correctness of the methodology. The precision of the results was determined by the obtained RSD value, which was 0.91% (less than 1%). Limit of detection (LOD) and limit of quantification (LOQ) took values of 0.1 and 1.0%, respectively. The range of measured concentrations was 0.01-20.0 mg/g. Conclusion . As a result of the studies, the method for sibutramine determination in dietary supplements with anorexigenic action was tested and can be used in quality control of dietary supplements., Competing Interests: The authors declare no overt and potential conflict of interest related to the publication of this article., (Copyright© GEOTAR-Media Publishing Group.)

A rapid screening method was developed to determine sibutramine and five derivatives in health food by high performance liquid chromatography-quadrupole/electrostatic orbitrap high-resolution mass spectrometry (HPLC-Q/Orbitrap HRMS). The sample was extracted with methanol via ultrasonic-assisted extraction coupled with high-speed centrifugation. Separation was performed on a Hypersil Gold column (100 mm×2.1 mm, 3 μm) by gradient elution with methanol/water (containing 0.15%(v/v) formic acid) as the mobile phase. The positive full mass scan/date-dependent MS 2 (Full MS/dd-MS 2 ) mode was used during MS detection, and quantification was achieved by resolution of the precursor mass. The analytes in the sample were separated, and accurate mass and MS 2 fragment ions were simultaneously attained within 8 min. The results indicated that the obtained mass accuracy errors of the six analytes were less than 1×10 -6 . Good linearities were obtained in the range of 0.5-20.0 μg/L, and all correlation coefficients were higher than 0.999. The limits of quantification were 25 μg/kg and the recoveries were in the range of 93.5%-103.5% with relative standard deviations of 1.5%-7.7%. This simple-pretreatment, rapid, accurate, high-sensitivity, and high-selectivity method can be used in the rapid screening and quantitative analysis of sibutramine and its derivatives in health food.

An innovative chromatographic analysis was developed for the determination of moniliformin (MON). Because of its ionic nature, MON is weakly retained in reversed-phase chromatography and the separation may be tricky. Nevertheless, this technique is normally used either with the formation of ion pairs or employing specific RP columns for polar compounds, or combining anion exchange and hydrophobic interactions. Hydrophilic interaction chromatography (HILIC) was also used, but a non-negligible peak tailing was observed. Besides its ionic nature, MON is a di-ketone and di-ketones, mainly β-di-ketones, can easily form complexes with lanthanide ions. Then, in this work the addition of lanthanide ions to the mobile phase was investigated, aiming at improving peak shape and MON separation. La 3+ , Tb 3+ or Eu 3+ aqueous solutions were used as mobile phase and MON was chromatographed using a LC-NH 2 column. The probable formation of coordination complexes lanthanide-MON in the HPLC mobile phase allowed to obtain a symmetrical peak shape and a satisfactory chromatographic separation by both mass spectrometry (MS/MS) and UV detection. Finally, a suitable extraction and purification method for MON determination in cereal samples was developed.

Maize (Zea mays), sorghum (Sorghum bicolor) and pearl millet (Pennisetum glaucum) are basic staple foods for many rural or poorer communities. These crops are susceptible to plant diseases caused by multiple species of Fusarium, some of which also produce mycotoxins, including fumonisins and moniliformin that are detrimental to both humans and domesticated animals. Eighteen potentially toxigenic Fusarium strains were isolated from maize (n = 10), sorghum (n = 7) and pearl millet (n = 1) growing in the same field in Nigeria. The 17 strains from maize and sorghum were all F. proliferatum and the one strain from pearl millet was F. pseudonygamai. Under conducive conditions, the 17 F. proliferatum strains produced fumonisins, 11 in relatively large quantities (700-17,000 mg total fumonisins, i.e., FB 1  + FB 2  + FB 3 /kg culture material), and six at <45 mg/kg. Ten F. proliferatum strains produced >100 mg of moniliformin per kg culture material with a maximum of 8900 mg/kg culture material. All strains could use all grains for growth and toxin production, regardless of the host from which they were isolated. Isolates varied in the amount of toxin produced on each substrate, with toxin production a property of the strain and not the host from which the strain was recovered. However, the extent to which a toxin-producing phenotype could be altered by the grain on which the fungus was grown is consistent with subtle genetic × environment interactions that require a larger data set than the one presented here to rigorously identify. In conclusion, there is significant variation in the ability of strains of F. proliferatum to produce fumonisins and moniliformin on maize, sorghum and millet. If the amount of toxin produced on the various grains in this study reflects real-world settings, e.g., poor storage, then the consumers of these contaminated grains could be exposed to mycotoxin levels that greatly exceed the tolerable daily intakes., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Fusarium temperatum is an emerging maize pathogen that causes maize ear and stalk rot diseases and produces various mycotoxins including moniliformin, beauvericin, enniatins and fumonisin B1, which poses a potential risk to the human food or animal feed supply chains. Early detection of F. temperatum is crucial to prevent its derived mycotoxins from entering the food chain, and is also a useful tool in disease management practices. Here, we describe a loop-mediated isothermal amplification (LAMP) assay for rapid diagnosis of F. temperatum. The 28S ribosomal DNA sequences (28S rDNA) of F. temperatum were used to design a set of six primers. The reaction conditions were optimized for developing a fast assay with high specificity and sensitivity, and were able to detect the presence of less than 10 pg of target DNA per reaction within 60 min. Furthermore, the resulting amplicons were visualized by adding SYBR Green I to the reaction tubes. Suspected F. temperatum infected maize stalk samples collected from Yunnan province, China were identified using the developed LAMP assay. In conclusion, the method not only provides a rapid and specific screening for the existence of F. temperatum in a bulk of maize samples without using sophisticated equipment, but also is potentially useful for other agriculturally important toxigenic fungi., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: A survey on Fusarium species and moniliformin (MON) occurrence in sorghum grains collected from one of the main sorghum-producing areas of Argentina was conducted. Also, growth of F. thapsinum, one of the main sorghum pathogens, and MON production under different water activity (a w ) conditions on a sorghum-based medium were determined., Results: Infection of sorghum grains by Fusarium species ranged from 82.5 to 99%; closely related species F. verticillioides, F. thapsinum and F. andiyazi were the most frequently recovered, followed by F. proliferatum and F. subglutinans. By sequencing a portion of the translation elongation factor-1α (TEF-1α) gene and by maximum parsimony analysis, F. verticillioides and closely related species were identified as F. thapsinum, F. andiyazi and F. verticillioides. Species within the F. graminearum species complex (FGSC) were isolated in high frequency. Maximum growth rates of 12 F. thapsinum strains were obtained at 0.995 a w . All evaluated strains were able to produce MON at all a w values tested, but MON production was higher at 0.995-0.982 a w . MON was detected in 41% of the samples at levels ranging from 363.2 to 914.2 µg kg -1 ., Conclusion: This study provides new data on the occurrence of Fusarium species in sorghum grains destined for animal consumption in Argentina. The production of MON at different a w values showed that the toxin can be produced under field conditions. The risk to livestock exposed to daily low levels of MON associated with the toxin occurrence in the sorghum grains analyzed is unknown. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

This manuscript is a continuation of a recent study that described the use of fully integrated gas chromatography with direct deposition Fourier transform infrared detection and mass spectrometric detection (GC-FT-IR-MS) to identify and confirm the presence of sibutramine and AB-FUBINACA. The purpose of the current study was to employ the GC-FT-IR portion of the same instrument to quantify these compounds, thereby demonstrating the ability to identify, confirm, and quantify drug substances using a single GC-FT-IR-MS unit. The performance of the instrument was evaluated by comparing quantitative analytical figures of merit to those measured using an established, widely employed method for quantifying drug substances, high performance liquid chromatography with ultraviolet detection (HPLC-UV). The results demonstrated that GC-FT-IR was outperformed by HPLC-UV with regard to sensitivity, precision, and linear dynamic range (LDR). However, sibutramine and AB-FUBINACA concentrations measured using GC-FT-IR were not significantly different at the 95% confidence interval compared to those measured using HPLC-UV, which demonstrates promise for using GC-FT-IR as a semi-quantitative tool at the very least. The most significant advantage of GC-FT-IR compared to HPLC-UV is selectivity; a higher level of confidence regarding the identity of the analyte being quantified is achieved using GC-FT-IR. Additional advantages of using a single GC-FT-IR-MS instrument for identification, confirmation, and quantification are efficiency, increased sample throughput, decreased consumption of laboratory resources (solvents, chemicals, consumables, etc.), and thus cost.

The growing market of herbal remedies worldwide could pose severe problems to consumers' health due to the possible presence of potentially harmful, undeclared synthetic substances or analogues of prescription drugs. The present work shows a simple but effective approach to unequivocally identify synthetic anorectic compounds in allegedly 'natural' herbal extracts, by exploiting liquid chromatography/time of flight (Q-TOF LC/MS) technology coupled to liquid chromatography/triple quadrupole (LC-MS/MS) confirmation and quantitation. The procedure was applied to five tea herbal extracts and pills sold as coadjutant for weigh loss. The method exploited liquid-liquid sample extraction (LLE) and separation in a C18 (2.1mm×150mm, 1.8μm) column. QTOF acquisitions were carried out both in scan mode and all ion MS/MS mode and results were obtained after search against ad hoc prepared library. Sibutramine, 4-hydroxyamphetamine, caffeine and theophylline were preliminary identified samples. Confirmation and quantitation of the preliminary identified compounds were obtained in LC-MS/MS after preparation of appropriated standards. Sibutramine, caffeine and theophylline were finally confirmed and quantitate., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The amount of photolesions produced in DNA after exposure to physiological doses of ultraviolet radiation (UVR) can be estimated with high sensitivity and at low cost through an immunological assay, ELISA, which, however, provides only a relative estimate that cannot be used for comparisons between different photolesions such as cyclobutane pyrimidine dimer (CPD) and pyrimidine(6-4)pyrimidone photoproduct (64PP) or for analysis of the genotoxicity of photolesions on a molecular basis. To solve this drawback of ELISA, we introduced a set of UVR-exposed, calibration DNA whose photolesion amounts were predetermined and estimated the absolute molecular amounts of CPDs and 64PPs produced in mouse skin exposed to UVC and UVB. We confirmed previously reported observations that UVC induced more photolesions in the skin than UVB at the same dose, and that both types of UVR produced more CPDs than 64PPs. The UVR protection abilities of the cornified and epidermal layers for the lower tissues were also evaluated quantitatively. We noticed that the values of absorbance obtained in ELISA were not always proportional to the molecular amounts of the lesion, especially for CPD, cautioning against the direct use of ELISA absorbance data for estimation of the photolesion amounts. We further estimated the mutagenicity of a CPD produced by UVC and UVB in the epidermis and dermis using the mutation data from our previous studies with mouse skin and found that CPDs produced in the epidermis by UVB were more than two-fold mutagenic than those by UVC, which suggests that the properties of CPDs produced by UVC and UVB might be different. The difference may originate from the wavelength-dependent methyl CpG preference of CPD formation. In addition, the mutagenicity of CPDs in the dermis was lower than that in the epidermis irrespective of the UVR source, suggesting a higher efficiency in the dermis to reduce the genotoxicity of CPDs produced within it. We also estimated the minimum amount of photolesions required to induce the mutation induction suppression (MIS) response in the epidermis to be around 15 64PPs or 100 CPDs per million bases in DNA as the mean estimate from UVC and UVB-induced MIS.

A simple LC-MS/MS method was developed for the determination of moniliformin (MON) in maize and applied for the analysis of samples within the official food surveillance. The homogenized samples were extracted with acetonitrile/water 50/50 (v/v) which proved to have the highest extraction efficiency compared to other tested solvents. The centrifuged extracts were diluted with acetonitrile and were measured after chromatographic separation by HILIC (hydrophobic interaction liquid chromatography)-HPLC without any cleanup (dilute and shoot approach). The LOD and LOQ achieved by this procedure were 2.6 and 8.8 μg/kg, respectively. Thirty-nine samples of popcorn, maize meal, and semolina were collected in 2014 and 2015 at mills, cinemas, wholesale, and retail from the Bavarian market (Germany). The rate of contamination with MON was very high (97%) with levels ranging between the LOD and 847 μg/kg. The mean level was 118 μg/kg and the median, 39 μg/kg. The maximum value was detected in maize meal. The results are discussed with respect to possible health implications for the consumer.

A straightforward analytical method was developed and validated to determine the mycotoxin moniliformin in cereal-based foods. Moniliformin is extracted with water and quantified with liquid chromatography tandem mass spectrometry, and its presence confirmed with liquid chromatography-Orbitrap-high-resolution mass spectrometry. The method was validated for flour, bread, pasta and maize samples in terms of linearity, matrix effect, recovery, repeatability and limit of quantification. Quantification was conducted by matrix-matched calibration. Positive samples were confirmed by standard addition. Recovery ranged from 77 to 114% and repeatability from 1 to 14%. The limit of quantification, defined as the lowest concentration tested at which the validation criteria of recovery and repeatability were fulfilled, was 10 μg/kg. The method was applied to 102 cereal-based food samples collected in the Netherlands and Germany. Moniliformin was not detected in bread samples. One of 22 flour samples contained moniliformin at 10.6 μg/kg. Moniliformin occurred in seven out of 25 pasta samples at levels around 10 μg/kg. Moniliformin (MON) was present in eight out of 23 maize products at levels ranging from 12 to 207 μg/kg.

Sibutramine may be illicitly included in herbal slimming foods and supplements marketed as "100% natural" to enhance weight loss. Considering public health and legal regulations, there is an urgent need for effective, rapid and reliable techniques to detect sibutramine in dietetic herbal foods, teas and dietary supplements. This research comprehensively explored, for the first time, detection of sibutramine in green tea, green coffee and mixed herbal tea using ATR-FTIR spectroscopic technique combined with chemometrics. Hierarchical cluster analysis and PCA principle component analysis techniques were employed in spectral range (2746-2656cm -1 ) for classification and discrimination through Euclidian distance and Ward's algorithm. Unadulterated and adulterated samples were classified and discriminated with respect to their sibutramine contents with perfect accuracy without any false prediction. The results suggest that existence of the active substance could be successfully determined at the levels in the range of 0.375-12mg in totally 1.75g of green tea, green coffee and mixed herbal tea by using FTIR-ATR technique combined with chemometrics., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Emerging Fusarium and Alternaria mycotoxins gain more and more interest due to their frequent contamination of food and feed, although in vivo toxicity and toxicokinetic data are limited. Whereas the Fusarium mycotoxins beauvericin, moniliformin and enniatins particularly contaminate grain and grain-based products, Alternaria mycotoxins are also detected in fruits, vegetables and wines. Although contamination levels are usually low (µg/kg range), higher contamination levels of enniatins and tenuazonic acid may occasionally occur. In vitro studies suggest genotoxic effects of enniatins A, A1 and B1, beauvericin, moniliformin, alternariol, alternariol monomethyl ether, altertoxins and stemphyltoxin-III. Furthermore, in vitro studies suggest immunomodulating effects of most emerging toxins and a reproductive health hazard of alternariol, beauvericin and enniatin B. More in vivo toxicity data on the individual and combined effects of these contaminants on reproductive and immune system in both humans and animals is needed to update the risk evaluation by the European Food Safety Authority. Taking into account new occurrence data for tenuazonic acid, the complete oral bioavailability, the low total body clearance in pigs and broiler chickens and the limited toxicity data, a health risk cannot be completely excluded. Besides, some less known Alternaria toxins, especially the genotoxic altertoxins and stemphyltoxin III, should be incorporated in risk evaluation as well., Competing Interests: The authors declare no conflict of interest.

A simple, fast and selective stability indicating liquid chromatographic method has been described for the simultaneous determination of squaric acid dibutyl ester and its impurities. The chromatographic separation was achieved on a C 2 column (250mm×4.6mm i.d., 5μm) using a mobile phase consisting of 0.15% phosphoric acid - acetonitrile - methanol (30:60:10, v/v/v). Isocratic elution was performed at a flow rate of 1.0mLmin -1 . The analytes were detected by UV at 252nm. The method was validated according to the ICH guidelines and satisfactory results were obtained. The specificity of the developed method was tested using forced degradation solutions of the drug substance. Characterization of squaric acid dibutyl ester and its forced degradation products was achieved by coupling mass spectrometry (MS) to the liquid chromatographic (LC) system. The method was successfully applied for quality control purposes including assay and determination of related compounds as required by regulatory guidelines to ensure its safety and efficacy since no monograph is available in official compendia., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The fungal and multi-mycotoxin profiles of groundnuts sold in domestic markets in Nigeria as well as the associated risk to consumers were assessed in the present study. Four hundred fungal isolates representing mainly Aspergillus [58.6%: Aspergillus section Flavi (37.1%) and A. niger-clade (21.5%)], Penicillium (40.9%) and Fusarium (0.5%) were isolated from 82 (97.6%, n=84) groundnut samples collected from four agro-ecological zones (AEZs) of Nigeria. The incidence of aflatoxin-producing A. flavus isolates (71%) was significantly (p<0.05) higher in the groundnuts than that of the non-aflatoxigenic isolates (29%). Fifty-four fungal metabolites [including aflatoxins (AFB 1 , AFB 2 , AFG 1 , AFG 2 and AFM 1 ), beauvericin (BEAU), cyclopiazonic acid (CPA), moniliformin, nivalenol and ochratoxin A] and four bacterial metabolites were detected in the groundnuts by liquid chromatography tandem mass spectrometry. Aflatoxins (39%; max: 2076μg/kg; mean: 216μg/kg) were detected in more samples than any other mycotoxin. About 25, 23 and 14% of the samples respectively were above the 2μg/kg AFB 1 , 4 and 20μg/kg total aflatoxin limits of the European Union and US FDA respectively. The mean margins of exposure of AFB 1 and total aflatoxins for adult consumers were 1665 and 908, respectively, while mean estimated daily intake values for infants, children and adults were <0.1% for BEAU and 4% for CPA. Consumers of mycotoxin contaminated groundnuts in Nigeria may therefore be at a risk of liver cancer in addition to other combinatory effects of mycotoxin/metabolite cocktails. There is need for increased targeted interventions in the groundnut value chain in Nigeria for public health benefits., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Slimming supplements were popularly sold online driven by the increasement of obesity and the development of social networking platform. However, events of drug abuse in slimming supplements were also frequently reported. In this study, a graphene tip solid-phase extraction (Gtip SPE) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was established for determining fenfluramine, phenolphthalein, bumetanide, and sibutramine in slimming supplements. It was validated in terms of linearity (0.9985-0.9995), LOD (1.8ngmL -1 ), LOQ (5.6ngmL -1 ), intra-day precision (<5.1%), inter-day precision (<7.3%), and recovery (82.9-95.2%). Sibutramine is the most commonly used drug, which was detected in Bihais, Galong, and Aolist, with content 12.4, 3.6, 20.3mgg -1 , respectively. Phenolphthalein was also found with content lower than 5.2mgg -1 . The successful application of Gtip SPE and UPLC-MS/MS method indicated its advantage in analyzing low level of contaminates resulted from violation of regulation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

18 F-fluciclovine ( trans -1-amino-3- 18 F-fluorocyclobutanecarboxylic acid) is an amino acid positron emission tomography (PET) tracer used for cancer staging (e.g., prostate and breast). Patients scheduled to undergo amino acid-PET are usually required to fast before PET tracer administration. However, there have been no reports addressing whether fasting improves fluciclovine-PET imaging. In this study, the authors investigated the influence of fasting on fluciclovine-PET using triple-tracer autoradiography with 14 C-fluciclovine, [5,6-³H]-2-fluoro-2-deoxy-d-glucose (³H-FDG), and 99m Tc-hydroxymethylene diphosphonate ( 99m Tc-HMDP) in a rat breast cancer model of mixed osteolytic/osteoblastic bone metastases in which the animals fasted overnight. Lesion accumulation of each tracer was evaluated using the target-to-background (muscle) ratio. The mean ratios of 14 C-fluciclovine in osteolytic lesions were 4.6 ± 0.8 and 2.8 ± 0.6, respectively, with and without fasting, while those for ³H-FDG were 6.9 ± 2.5 and 5.1 ± 2.0, respectively. In the peri-tumor bone formation regions (osteoblastic), where 99m Tc-HMDP accumulated, the ratios of 14 C-fluciclovine were 4.3 ± 1.4 and 2.4 ± 0.7, respectively, and those of ³H-FDG were 6.2 ± 3.8 and 3.3 ± 2.2, respectively, with and without fasting. These results suggest that fasting before 18 F-fluciclovine-PET improves the contrast between osteolytic and osteoblastic bone metastatic lesions and background, as well as 18 F-FDG-PET.

Previous studies have established that 2-alkylcyclobutanones (2-ACBs) are unique radiolytic products in lipid-containing foods that could only be formed through exposure to ionizing radiation, but not by any other means of physical/heat treatment methods. Therefore, 2-ACBs are currently the marker molecules required by the European Committee for Standardization to be used to identify foods irradiated with ionizing irradiation. Using a spectrum of state-of-the-art analytical instruments, we present in this study for the first time that the generation of 2-ACBs was also possible when fatty acids and triglycerides are exposed to a non-ionizing, short-wavelength ultraviolet (UV-C) light source. An irradiation dosage-dependent formation of 2-ACBs was also observed in UV-C irradiated fatty acids, triglycerides, corn oil, and pork samples. With UV-C irradiation becoming an increasingly common food treatment procedure, it is anticipated that the results from this study will alert food scientists and regulatory officials to a potential new source for 2-ACBs., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The purpose of this study was to develop an LC-tandem MS method for the simultaneous detection of common synthetic drugs as adulterants in natural and herbal slimming products. Sixteen drugs belonging to a wide range of pharmaceutical classes were studied. Included in the list of drugs were anorexics, anxiolytics, antidepressants, diuretics, laxatives, and stimulants. The method used a C18 column (4.6 × 50 mm and 1.8 μm particle size). Separation of the drugs was achieved by gradient elution using 4 mM ammonium formate in water + 0.1% formic acid as the aqueous component and 4 mM ammonium formate in methanol + 0.1% formic as the organic component of the mobile phase. As not all of the analytes ionized in the positive mode, the mass spectrometer was operated in the electrospray ionization mode with polarity switching. The samples were extracted with methanol and the use of 50% acetonitrile in water and 50% methanol in water were investigated as diluents for injection into the LC-MS system. Utilizing both diluents, the validation parameters including accuracy, precision, LOD, and LOQ were assessed. The validation results and utilization of the method to analyze a variety of weight-loss supplements indicate that the two diluents give similar results and can be used interchangeably. This knowledge provides the user with the option of selecting either diluent for sample preparation depending on the sample matrix without having to revalidate the method. The method was applied to the analysis of weight-loss supplements available in local pharmacies, herbal pharmacies, and over the Internet.

Irradiation of food products and ingredients must be indicated by proper labeling. This study evaluated the appropriateness of the European Standard EN 1785:2003 for the detection of 2-alkylcyclobutanones, which are radiolysis products of fatty acids, in cashew nuts and nutmeg and confirmed its suitability to detect irradiation of cashew nut samples at average absorbed doses of 1 kGy and above. An alternative method was developed, which is based on matrix solid phase dispersion and subsequent separation and detection of oxime derivatives of 2-alkylcyclobutanones by high performance-high resolution mass spectrometry. It is more rapid, less resource consuming, and more sensitive than EN 1785:2003. This method allowed detection of 2-alkylcyclobutanones in cashew nuts irradiated at 100 Gray and in nutmeg irradiated at 400 Gray. None of the 26 cashew nut and 14 nutmeg samples purchased in different EU Member States contained traces of 2-alkylcyclobutanones., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The aim of the project was to produce updated information during 2005-14 on the Fusarium species found in Finnish cereal grains, and the toxins produced by them, as the last comprehensive survey study of Fusarium species and their toxins in Finland was carried out at the turn of the 1960s and the 1970s. Another aim was to use the latest molecular and chemical methods to investigate the occurrence and correlation of Fusarium species and their mycotoxins in Finland. The most common Fusarium species found in Finland in the FinMyco project 2005 and 2006 were F. avenaceum, F. culmorum, F. graminearum, F. poae, F. sporotrichioides and F. langsethiae. F. avenaceum was the most dominant species in barley, spring wheat and oat samples. The occurrence of F. culmorum and F. graminearum was high in oats and barley. Infection by Fusarium fungi was the lowest in winter cereal grains. The incidence of Fusarium species in 2005 was much higher than in 2006 due to weather conditions. F. langsethiae has become much more common in Finland since 2001. F. graminearum has also risen in the order of importance. A highly significant correlation was found between Fusarium graminearum DNA and deoxynivalenol (DON) levels in Finnish oats, barley and wheat. When comparing the FinMyco data in 2005-06 with the results of the Finnish safety monitoring programme for 2005-14, spring cereals were noted as being more susceptible to infection by Fusarium fungi and the formation of toxins. The contents of T-2 and HT-2 toxins and the frequency of exceptionally high DON concentrations all increased in Finland during 2005-14. Beauvericin (BEA), enniatins (ENNs) and moniliformin (MON) were also very common contaminants of Finnish grains in 2005-06. Climate change is leading to warmer weather, and this may indicate more changes in Finnish Fusarium mycobiota and toxin contents and profiles in the near future.

The effect of water activity (aw = 0.95, 0.98 and 0.995), temperature (15, 25 and 30°C), incubation time (7, 14, 21 and 28 days), and their interactions on growth and moniliformin (MON), beauvericin (BEA), fusaproliferin (FUS) and fumonisin B1 (FB1) production by two strains of Fusarium temperatum isolated from Argentinean maize were determined in vitro on sterile layers of maize grains. The results showed that there was a wide range of conditions for growth and mycotoxins production by F. temperatum. Both strains were found to grow faster with increasing aw and at 30°C. In relation to mycotoxin production, the two strains produced more FUS than the other mycotoxins regardless of aw or temperature evaluated (maximum = 50,000 μg g(-1)). For FUS, MON and BEA, the maximum levels were observed at 0.98 aw and 30°C (50,000, 5000 and 2000 μg g(-1) respectively). The lowest levels for these three mycotoxins were detected at 15°C and 0.95 aw (1700 and 100 μg g(-1) for FUS and MON respectively), and at 0.98 aw (400 μg g(-1) for BEA). The maximum levels of FB1 were produced at 15°C and 0.98 aw (1000 μg g(-1)). At all aw and temperatures combinations evaluated there was an increase in toxin concentrations with time incubation. The maximum levels were detected at 21 days. Statistical analyses of aw, temperature, incubation time, and the two- and three-way interactions between them showed significant effects on mycotoxins production by F. temperatum. For its versatility on growth and mycotoxin production, F. temperatum represents a toxicological risk for maize in the field and also during grain storage.

Background: Weight-loss medicines, including crude drugs and herbal supplements disguised as diet-aid products, are readily obtainable and distributed widely, especially in Southeast Asia. Even if such products are unapproved or prescription-only medicines, consumers can purchase them through an agency or directly on the Internet. We evaluated the quality and safety of herbal products purchased on the Internet to reveal their influence on public health., Methods: Diet-aid products containing Bukuryo (Poria sclerotium), Bakumondo (Ophiopogonis tuber), or Daio (rhubarb rhizome) were purchased through websites that did not provide physical addresses or which advertised misleading medicines (e.g., unapproved Cialis 100 mg tablets, Viagra 100 mg tablets) on websites. We carefully noted details in the descriptions on package inserts or accompanying product characteristics and analyzed the ingredients using qualitative and quantitative methods, namely high-performance liquid chromatography equipped with a photodiode array detector. We requested the respective manufacturers to authenticate their products through a structured questionnaire., Results: We purchased 15 items from 15 Internet sites and imported all 15 items to Japan. One item stated to contain rhubarb rhizome was identified as a prescription medicine; the others were dietary supplements and not medicines. Even though we did not analyze the constituents of all crude drugs, we found some active ingredients in the items. Sibutramine was detected in items confirmed to be supplements, including those containing Poria sclerotium and Ophiopogonis tuber. Each capsule contained ≈ 12 mg of sibutramine, which is the daily dose for anti-obesity medicines. Sibutramine is not approved for use in Japan and its sale has been suspended in Europe and the USA owing to serious adverse effects on the circulatory system., Conclusions: Our findings indicate that dietary supplements containing injurious ingredients are distributed to Japanese consumers and potentially to a broader international audience, and that purchasing them through unreliable websites bears potential health risks. To avoid potential adverse events, there should be adequate alerts about the risks of taking products without appropriate indications.

A novel biosensor regulated by the rotator of F0F1-ATPase was developed to analyze 2-dodecylcyclobutanone (2-DCB) to detect γ-ray irradiated beef rapidly. The biosensor was assembled by conjugating 2-DCB monoclonal antibodies with the "rotator" ε-subunit of F0F1-ATPase within chromatophores through an ε-subunit monoclonal antibody-biotin-avidin-biotin linker. The limit of detection (LOD) of 2-DCB was approximately 10(-8) μg/mL. The recovery ratio of 2-DCB from ground beef patties ranged from 75.1% to 116.4%. The intra-assay and inter-assay coefficients of variation were both <15.0%. The proposed method was validated by gas chromatography-mass spectrometry with high correlation. The biosensor was used to detect 2-DCB in ground beef patties with different fat contents (10%, 20%, and 30%) irradiated at 0.5, 1.0, 3.0, 5.0, and 7.0 kGy. The 2-DCB concentration linearly increased with the radiation dose in all the beef samples. 2-DCB concentration increased with fat levels in the three samples., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

There is an increasing demand for herbal medicines in weight loss treatment. Some synthetic chemicals, such as sibutramine (SB), have been detected as adulterants in herbal formulations. In this study, two strategies using near infrared (NIR) spectroscopy have been developed to evaluate potential adulteration of herbal medicines with SB: a qualitative screening approach and a quantitative methodology based on multivariate calibration. Samples were composed by products commercialized as herbal medicines, as well as by laboratory adulterated samples. Spectra were obtained in the range of 14,000-4000 per cm. Using PLS-DA, a correct classification of 100% was achieved for the external validation set. In the quantitative approach, the root mean squares error of prediction (RMSEP), for both PLS and MLR models, was 0.2% w/w. The results prove the potential of NIR spectroscopy and multivariate calibration in quantifying sibutramine in adulterated herbal medicines samples., (© 2015 American Academy of Forensic Sciences.)

To counter the growth of herbal medicines adulterated with pharmaceuticals crossing borders, rapid, inexpensive and non-destructive analytical techniques, that can handle complex matrices, are required. Since mid-infrared (MIR), near infrared (NIR) and Raman spectroscopic techniques meet these criteria, their performance in identifying adulterants in seized weightloss herbal medicines is definitively determined. Initially a validated high pressure liquid chromatography methodology was used for reference identification and quantification of the adulterants sibutramine H2O·HCl, fenfluramine HCl and phenolphthalein. Of 38 products, only sibutramine and phenolphthalein were detected by HPLC. The spectroscopic measurements showed Raman was ill-suited due to sample burning and emission while NIR lacked adulterant selectivity. Conversely, MIR demonstrated apt identification performance, which manifested as spectrally meaningful separation based on the presence and type of adulterant during principal component analysis (test set validated). Partial least squares regression models were constructed from the MIR training sets for sibutramine and phenolphthalein - both models fitted the training set data well. Average test set prediction errors were 0.8% for sibutramine and 2.2% for phenolphthalein over the respective concentration ranges of 1.7-11.7% and 0.9-34.4%. MIR is apposite for the screening of anorectic and laxative adulterants and is the most viable technique for wider adulterant screening in herbal medicines., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The availability of a simple chemical precipitation workflow aided by targeted and untargeted mass spectrometry would provide an accurate diagnostic platform for the direct determination of moniliformin in cereals for food safety control. In-house method validation was performed at six concentration levels of 8, 40, 80, 200, 400, and 600 ng g(-1) in cereal flours of wheat, corn, rye, oats and barley. Spiking experiments were made at three concentration levels of 20, 40 and 100 ng g(-1). Protein precipitation and "PHREE" column cleanup strategy provided recoveries of 81-108% for all cereals matrices using external calibrants. "PHREE" purification provided significant (p < 0.05) ion signal enhancement reduction advantage for all matrices except corn flour. Moniliformin underwent significant (p < 0.05) degradation over 2 weeks when prepared in acidified water. A simple, low-cost and fit-for-purpose procedure for the identification and quantitation of moniliformin in cereals becomes available to support prospective regulatory function., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Purpose: Androgens play a crucial role in prostate cancer progression, and trans-1-amino-3-[(18)F]fluorocyclobutanecarboxylic acid (anti-[(18) F]FACBC) are used for visualization of prostate cancer. We examined the effect of androgen on the expression of amino acid transporters related to anti-[(18)F]FACBC transport and uptake of trans-1-amino-3-fluoro-[1-(14)C]cyclobutanecarboxylic acid (anti-[(14)C]FACBC)., Procedures: Expression of amino acid transporters and uptake of anti-[(14)C]FACBC in androgen receptor (AR)-positive LNCaP and AR-negative DU145 human prostate cancer cells cultured with/without 5α-dihydrotestosterone (DHT) and the effect of bicalutamide, an AR antagonist, on DHT-associated changes were investigated., Results: DHT stimulated the expression of amino acid transporters ASCT2, SNAT5, 4F2 heavy chain, and LAT3 in LNCaP but not in DU145 cells. Anti-[(14)C]FACBC uptake was enhanced, in a DHT-dependent manner, in LNCaP cells only., Conclusions: DHT enhanced the expression of ASCT2, the transporter responsible for anti-[(18)F]FACBC uptake, thereby increasing anti-[(14)C]FACBC uptake in AR-positive LNCaP cells. Androgen-mediated induction may contribute to the distinct anti-[(18)F]FACBC accumulation pattern in prostate cancer.

Sibutramine is one of the most occurring adulterants encountered in dietary supplements with slimming as indication. These adulterated dietary supplements often contain a herbal matrix. When customs intercept these kind of supplements it is almost impossible to discriminate between the legal products and the adulterated ones, due to misleading packaging. Therefore in most cases these products are confiscated and send to laboratories for analysis. This results inherently in the confiscation of legal, non-adulterated products. Therefore there is a need for easy to use equipment and techniques to perform an initial screening of samples. Attenuated total reflectance-infrared (ATR-IR) spectroscopy was evaluated for the detection of sibutramine in adulterated dietary supplements. Data interpretation was performed using different basic chemometric techniques. It was found that the use of ATR-IR combined with the k-Nearest Neighbours (k-NN) was able to detect all adulterated dietary supplements in an external test set and this with a minimum of false positive results. This means that a small amount of legal products will still be confiscated and analyzed in a laboratory to be found negative, but no adulterated samples will pass the initial ATR-IR screening., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The isomeric truxillines are a group of minor alkaloids present in all illicit cocaine samples. The relative amount of truxillines in cocaine is indicative of the variety of coca used for cocaine processing, and thus, is useful in source determination. Previously, the determination of isomeric truxillines in cocaine was performed with a gas chromatography/electron capture detection method. However, due to the tedious sample preparation as well as the expense and maintenance required of electron capture detectors, the protocol was converted to a gas chromatography/flame-ionization detection method. Ten truxilline isomers (alpha-, beta-, delta-, epsilon-, gamma-, omega, zeta-, peri-, neo-, and epi-) were quantified relative to a structurally related internal standard, 4',4″-dimethyl-α-truxillic acid dimethyl ester. The method was shown to have a linear response from 0.001 to 1.00 mg/mL and a lower detection limit of 0.001 mg/mL. In this method, the truxillines are directly reduced with lithium aluminum hydride and then acylated with heptafluorobutyric anhydride prior to analysis. The analysis of more than 100 cocaine hydrochloride samples is presented and compared to data obtained by the previous methodology. Authentic cocaine samples obtained from the source countries of Colombia, Bolivia, and Peru were also analyzed, and comparative data on more than 23,000 samples analyzed over the past 10 years with the previous methodology is presented., (Published by Elsevier B.V.)

2-Dodecylcyclobutanone (2-DCB) is one of the major 2-alkylcyclobutanones (2-ACBs) that are uniquely formed when triglycerides are exposed to ionizing radiation and is being widely used as marker signature for identifying irradiated food. Current methods for 2-DCB extraction are either time-/solvent-consuming or involve the use of expensive extraction instruments. We have developed in this study an efficient extraction method for extracting 2-DCB from γ-irradiated food by direct extraction using acetonitrile. In comparison with the European standard method for irradiated food identification, our method reduced the per sample organic solvent consumption from 450 to 30mL per sample and shortened sample processing time from 8h to 30min. The assay combining direct acetonitrile extraction, precolumn derivatization with hydroxylamine, and LC-MS/MS analysis was validated with irradiated chicken as positive control, and applied in surveillance of irradiated food in Hong Kong., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Methods of analysis for 2-dodecylcyclobutanone (2-DCB) using gas chromatography with mass spectrometric detection (GC-MS), liquid chromatography with time-of-flight mass spectrometric detection (LC-TOF-MS) and LC with tandem MS (MS/MS) detection have been developed and optimised for maximum sensitivity to allow very low irradiation doses to be detected. The LC-MS/MS method, following derivatisation with 2,4-dinitrophenylhydrazine, was found to be the most sensitive technique and was used to determine the amount of 2-DCB formed from the model compounds palmitic acid, glyceryl tripalmitate and 1,3-dipalmitoyl-2-oleoylglycerol irradiated over a range of doses by two different irradiation sources (gamma and electron beam). The model compounds were also treated with a number of non-irradiation based processing techniques including heating in the presence and absence of oxygen, light, and redox active metal salts, in a conventional oven, microwave oven and pressure cooker. No 2-DCB was detected in any of the processed non-irradiated model compounds, reaffirming the hypothesis that 2-DCB is a unique radiolytic product that can be used as a marker of irradiation in foodstuffs., (Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.)

A new method has been developed using flow injection tandem mass spectrometry to semi-quantitatively screen for weight loss drugs, including sibutramine, N-desmethylsibutramine, N-didesmethylsibutramine, and phenolphthalein in dietary supplements. Positive identification of these drugs in samples was further confirmed and quantified by liquid chromatography tandem mass spectrometry. The degradation products of sibutramine were observed and identified by LC-MS/MS which include N-desmethylsibutramine, N-didesmethylsibutramine, N-formyldesmethylsibutramine, and N-formyldidesmethylsibutramine., (Published by Elsevier B.V.)

The adulteration of herbal supplements is of growing importance, especially when they contain undeclared compounds like sibutramine that are unsafe drugs. Sibutramine was withdrawn from US and European markets in 2010. In this study, an HPTLC-UV densitometric method was developed for the quantification of sibutramine in herbal diet foods. Sample extracts were directly applied onto HPTLC silica gel plates and separated with a mobile phase made of a toluene-methanol mixture. Sibutramine was quantified at 225 nm and its unequivocal identification was confirmed by MS using a TLC-MS interface. During two surveys, 52 weight loss supplements obtained via the Internet were screened. Half of those were adulterated with sibutramine at amounts reaching up to 35 mg per capsule. The results of this validated HPTLC method were compared with those obtained by HPLC-UV and HPLC-MS/MS. The results were not significantly different with the three methods.

Because of the rapid growth in dietary supplement availability and public concern for weight control, the investigation of foods and various dietary supplements illegally adulterated with weight loss compounds has become increasingly important. A total of 29 weight loss compounds, including sennoside, sibutramine, ephedrine and their analogues, found to be adulterated in foods and dietary supplements were simultaneously examined by LC-MS/MS. The 188 samples were collected between 2009 and 2012 in South Korea, and method validation was performed to determine the adulterants to the weight loss compounds. LODs, LOQs and linearity ranged from 0.03 to 7.5 ng ml⁻¹, from 0.08 to 30.00 ng ml⁻¹, and from 0.990 to 0.999, respectively. The results showed that nine weight loss compounds, namely bisacodyl, desmethylsibutramine, didesmethylsibutramine, ephedrine, fluoxetine, pseudoephedrine, sennoside A, sennoside B and sibutramine, were detected in 62 of all collected samples and were found in order of frequency as follows: sibutramine, 25.7%; sennoside A, 22.9%; sennoside B, 20.0%; fluoxetine, 8.6%; desmethylsibutramine, 7.1%; bisacodyl, ephedrine, and pseudoephedrine, 4.3%; and didesmethylsibutramine, 2.9%. Sibutramine, which was the most frequently found adulterant, ranged in levels from 0.03 to 132.40 mg g⁻¹ (2010), from 0.88 to 76.2 mg g⁻¹ (2011), and from 0.07 to 0.24 mg g⁻¹ (2012). Although the concentrations of most compounds ranged widely, some compounds such as bisacodyl and fluoxetine were found at high concentrations in several samples.

2-Alkylcyclobutanones (2-ACBs) have long been considered as unique radiolytic products that can be used as indicators for irradiated food identification. A recent report on the natural existence of 2-ACB in non-irradiated nutmeg and cashew nut samples aroused worldwide concern because it contradicts the general belief that 2-ACBs are specific to irradiated food. The goal of this study is to test the natural existence of 2-ACBs in nut samples using our newly developed liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with enhanced analytical sensitivity and selectivity ( Ye , Y. ; Liu , H. ; Horvatovich , P. ; Chan , W. Liquid chromatography-electrospray ionization tandem mass spectrometric analysis of 2-alkylcyclobutanones in irradiated chicken by precolumn derivatization with hydroxylamine . J. Agric. Food Chem. 2013 , 61 , 5758 - 5763 ). The validated method was applied to identify 2-dodecylcyclobutanone (2-DCB) and 2-tetradecylcyclobutanone (2-TCB) in nutmeg, cashew nut, pine nut, and apricot kernel samples (n = 22) of different origins. Our study reveals that 2-DCB and 2-TCB either do not exist naturally or exist at concentrations below the detection limit of the existing method. Thus, 2-DCB and 2-TCB are still valid to be used as biomarkers for identifying irradiated food.

A highly sensitive and specific competitive indirect enzyme-linked immunosorbent assay (ciELISA) based on monoclonal antibodies was developed for the detection of 2-dodecylcyclobutanone (2-DCB), a chemical marker for irradiated lipid-containing foods. 2-Oxocyclobutane undecanoic acid was used as an alternative to 2-DCB and conjugated to BSA and OVA via a conventional carbodiimide condensation reaction to prepare the immunogen and the coating antigen for 2-DCB. The monoclonal antibody against 2-DCB was obtained using the hybridoma technique, with a high specificity and low cross-reactivity for 2-tetradecylcyclobutanone (2-TCB; <8%) and other structurally related compounds (<0.1%). The ciELISA method was applicable at optimal experimental conditions of 0.001-100 μg/mL 2-DCB in a buffer solution, with an IC50 value of 0.25 μg/mL and a limit of detection (defined as the IC20) of approximately 0.004 μg/mL. The recovery efficiency of 2-DCB from ground beef patties ranged from 84.4 to 109.8%. The intra-assay and interassay coefficients of variation were <10.0 and <12.0%, respectively. The proposed method was validated by gas chromatography-mass spectrometry with high correlation. The same method was used to detect 2-DCB in ground beef patties irradiated at 0.5, 1.0, 3.0, 5.0, and 7.0 kGy; the 2-DCB concentration linearly increased with the radiation dose.

Food irradiation is a common preservation method that is used in many countries. The ability to identify irradiated food is important for assuring compliance with regulatory policies, such as food labeling requirements, and for informed consumer choice. There is thus a significant demand for analytical methods of high sensitivity and selectivity to identify irradiated food, especially for foods subjected to low-dose irradiation and for processed or composite foods that contain small quantities of irradiated ingredients. 2-Alkylcyclobutanones (2-ACBs) are uniquely formed during food irradiation and have been adopted by the European Committee for Standardization as signature biomarkers for the identification of irradiated foods. We now report the development of a novel assay for quantification of 2-ACBs in γ-irradiated food by liquid extraction of fat content followed by precolumn derivatization and liquid chromatography-tandem mass spectrometric (LC-MS/MS) detection. Precolumn derivatization with hydroxylamine introduced a polar functional group into the otherwise nonpolar 2-ACBs, which greatly enhanced ESI-MS response. The method was validated for extraction efficiency, precision, accuracy, and detection limit. In comparison with the current GC-MS based European official method (EN1785:2003) for 2-ACBs determination, our new LC-MS/MS method offers a more efficient sample processing protocol with reduced solvent consumption. More importantly, the combination of chemical derivatization and LC-MS/MS detection significantly enhanced the analytical sensitivity of the method, which allows confident identification of food irradiated with as little as 10 Gy. To the best of our knowledge, this is the first report of 2-ACB determination by LC-MS/MS and the first analytical method allowing confident identification of irradiated food at dosage of down to 10 Gy.

The combination of (1)H NMR lipid profiling with multivariate analysis was applied to differentiate irradiated and non-irradiated beef. Two pattern recognition chemometric procedures, stepwise linear discriminant analysis (sLDA) and artificial neural networks (ANNs), provided a successful discrimination between the groups investigated. sLDA allowed the classification of 100% of the samples into irradiated or non-irradiated beef groups; the same result was obtained by ANNs using the 1 kGy irradiation dose as discriminant value suggested by the network. Furthermore, sLDA allowed the classification of 81.9% of the beef samples according to the irradiation dose (0, 2.5, 4.5 and 8 kGy). (1)H NMR lipid profiling, coupled with multivariate analysis may be considered a suitable and promising screening tool for the rapid detection of irradiated meat in official control of food., (Copyright © 2012 Elsevier Ltd. All rights reserved.)