Determination of phosphodiesterase 5 (PDE5)inhibitors in instant coffee premixes using liquid chromatography-high-resolution mass spectrometry (LC-HRMS)

© 2019 Elsevier B.V. As a widely consumed beverage, coffee tends to be a target for intentional adulteration. This study describes the application of modified quick, easy, cheap, effective, rugged, and safe (QuEChERS)coupled to liquid chromatography-high-resolution mass spectrometry (LC-HRMS)for simultaneous screening, identification, and quantification of undeclared phosphodiesterase 5 (PDE5)inhibitors in instant coffee premixes (ICPs). The mass spectrometer was operated in auto MS/MS acquisition for simultaneous MS and MS/MS experiments. Qualitative establishments from the suspected-target screening and targeted identification processes led to an unambiguous analyte assignment from the protonated molecule ([M+H]+)precursor ion which is subsequently used for quantification of 23 targeted PDE5 inhibitors. The analytical method validation covered specificity, linearity, range, accuracy, limit of detection (LOD), limit of quantification (LOQ), precisions, matrix effect (ME), and extraction recovery (RE). The specificity was established using the optimised chromatographic separation as well as the distinguishable [M+H]+ precursor ion. The linearity of each target analyte was demonstrated with a coefficient of determination (r2)of >0.9960 over the expected range of sample concentrations. The accuracy ranged from 88.1%–119.3% with LOD and LOQ of <70 ng/mL and 80 ng/mL, respectively. Excellent precisions were established within 0.4%–9.1% of the relative standard deviation. An insignificant ME within −5.2% to +8.7% was achieved using three different strategies of chromatography, sample extraction, and sample dilution. The RE was good for all target analytes within 84.7%–123.5% except for N-desethylacetildenafil at low (53.8%)and medium (65.1%)quality control levels. The method was successfully applied to 25 samples of ICPs where 17 of them were found to be adulterated with PDE5 inhibitors and their analogues. Further quantification revealed the total amount of these adulterants ranged from 2.77 to 121.64 mg per sachet

Data on the optimisation and validation of a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to establish the presence of phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes

© 2019 The Authors This paper presents the data on the optimisation and validation of a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to establish the presence of phosphodiesterase 5 (PDE5) inhibitors and their analogues as adulterants in instant coffee premixes. The method development data covered chromatographic optimisation for better analyte separation and isomeric resolution, mass spectrometry optimisation for high sensitivity and sample preparation optimisation for high extraction recovery (RE) and low matrix effect (ME). The validation data covered specificity, linearity, range, accuracy, limit of detection, limit of quantification, precisions, ME, and RE. The optimisation and validation data presented here is related to the article: “Determination of phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes using liquid chromatography-high-resolution mass spectrometry (LC-HRMS)” Mohd Yusop et al., 2019

Fluorescence polarisation for high-throughput screening of adulterated food products via phosphodiesterase 5 inhibition assay.

The surge in the consumption of food products containing herbal aphrodisiacs has driven their widespread adulteration. A rapid screening strategy is, therefore, warranted to curb this problem. This study established an enzyme inhibition assay to screen phosphodiesterase 5 (PDE5) inhibitors as adulterants in selected food products. Fluorescein-labelled cyclic-3',5'-guanosine monophosphate was utilised as substrates for the PDE5A1 enzyme, aided by the presence of nanoparticle phosphate-binding beads on their fluorescence polarisation. The sample preparation was optimised to improve the enzyme inhibition efficiency and applied to calculate the threshold values of six blank food matrices. The assay was validated using sildenafil, producing an IC50 of 4.2 nM. The applicability of the assay procedure was demonstrated by screening 55 distinct food samples. The results were subsequently verified using confirmatory liquid chromatography-high-resolution mass spectrometry (LC-HRMS) analysis. Altogether, 49 samples inhibited the PDE5 enzyme above the threshold values (75.7%-105.5%) and were registered as potentially adulterated samples. The remaining six samples were marked as nonadulterated with percentage inhibition below the threshold values (-3.3%-18.2%). The LC-HRMS analysis agreed with the assay results for all food products except for the instant coffee premix (ICP) samples. False-positive results were obtained for the ICP samples at 32% (8/25), due to possible PDE5 inhibition by caffeine. Contrarily, all other food samples were found to produce 0% (0/30) false-positive or false-negative results. The broad-based assay, established via a simple mix-incubate-read format, exhibited promising potential for high-throughput screening of PDE5 inhibitors in various food products, except those with naturally occurring phosphodiesterase inhibitors such as caffeine

Tetra­kis[μ-1,4-bis­(4,5-dihydro-1,3-oxazol-2-yl)benzene-κ2 N:N′]tetra­kis­(μ-methano­lato-κ2 O:O)bis­(μ-perchlorato-κ2 O:O′)tetra­copper(II) bis­(perchlorate)

The title tetra­nuclear CuII complex, [Cu4(C12H12N2O2)4(CH3O)4(ClO4)2](ClO4)2, is located around an inversion center. Each CuII atom is coordinated by two cis-O atoms from two bridging methano­late anions and two cis-N atoms from two bridging 1,4-bis­(4,5-dihydro-1,3-oxazol-2-yl)benzene (L) ligands in the basal plane, and is further coordinated by one O atom of the bridging perchlorate anion, forming a distorted square-pyramidal geometry. The Cu⋯Cu separations in the recta­ngular core are 2.9878 (11) and 6.974 (1) Å. In the asymmetric unit, there are two L ligands with a syn conformation. In one L ligand, the dihedral angles between the central benzene ring and the terminal 4,5-dihydro-1,3-oxazol-2-yl mean planes are 22.1 (4) and 33.1 (4)°, and in the other L ligand the corresponding dihedral angles are 29.3 (4) and 29.9 (4)°. The uncoordinated perchlorate anion is linked with the complex mol­ecules via weak C—H⋯O hydrogen bonds

Design Philosophy for Buildings’ Comfort-Level Performance

The data reported by Japan Meteorological Agency (JMA) show that the fatal casualties and severe injuries are due to heavy shaking during massive earthquakes. Current earthquake-resistant building standards do not include comfort-level performance. Hence, a new performance design philosophy is proposed in this research to evaluate the quantitative effect of earthquake-induced shaking in a building. The earthquake-induced response accelerations in a building are analysed, and the response accelerations related with the characteristic property of the building are used to evaluate the number of Seismic Intensity Level (SIL). To show the indispensability of the newly proposed comfort-level design philosophy, numerical simulations are conducted to evaluate the comfort level on different floors in a building. The results show that the evaluation of residents’ comfort levels should be considered in the current earthquake-resistant building design codes

Poly[(μ3-quinoline-6-carboxyl­ato-κ3 N:O:O′)silver(I)]

In the title coordination polymer, [Ag(C10H6NO2)]n, the AgI cation is coordinated by two O atoms and one N atom from three 6-quinoline­carboxyl­ate anions in a distorted T-shaped AgNO2 geometry, in which the O—Ag—O angle is 160.44 (9)°. The 6-quinoline­carboxyl­ate anion bridges three Ag+ cations, forming a nearly planar polymeric sheet parallel to (101). The distance between Ag+ cations bridged by the carboxyl group is 2.9200 (5) Å. In the crystal, π–π stacking is observed between parallel quinoline ring systems, the centroid–centroid distance being 3.7735 (16) Å

Isolation and identification of an isomeric sildenafil analogue as an adulterant in an instant coffee premix

Abstract The proliferation of adulterated health foods and beverages in the market demands a comprehensive analytical strategy to identify the adulterants, particularly those of isomeric phosphodiesterase 5 (PDE5) inhibitors. An instant coffee premix (ICP) purchased from an online retailer was flagged for suspected adulteration through PDE5 inhibition assay. The ICP was then analysed using suspected-target and non-targeted screenings of a liquid chromatography-quadrupole time-of-flight mass spectrometry. Based on these findings, a PDE5 inhibitor initially assigned as compound X was isolated from the ICP by employing a liquid chromatography-diode array detection before its structural elucidation with liquid chromatography-ultraviolet (LC-UV) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. The suspected-target screening matched the protonated molecule ([M?+?H]+) precursor ion of compound X at m/z 499.2310 with two suspected analytes that are structural isomers of one another. The fragmentation patterns of compound X were comparable to those analogues in the dithiocarbodenafil group through the non-targeted screening. These findings, complemented by the LC-UV and NMR spectroscopy data, together with the chromatographic separation of related structural isomers, conclude the identity of compound X. To our best knowledge, this is the first study to report the presence of 3,5-dimethylpiperazinyl-dithiodesmethylcarbodenafil in an ICP sample. Key points The herbal-based male sexual performance products? lucrative market has instigated their rampant adulteration, particularly with PDE5 inhibitors. The adulterated products may also contain analogues of the approved PDE5 inhibitors, which usually passed into the market undetected as they are not included in the routine targeted screening procedure. The present study detected, isolated, and identified an isomeric sildenafil analogue from an instant coffee premix sample using rapid qualitative assay and comprehensive analytical analysis. This paper highlighted the applicability of the established strategies for routine casework, particularly in a forensic drug testing laboratory

Cell sorting in a Petri dish controlled by computer vision.

Fluorescence-activated cell sorting (FACS) applying flow cytometry to separate cells on a molecular basis is a widespread method. We demonstrate that both fluorescent and unlabeled live cells in a Petri dish observed with a microscope can be automatically recognized by computer vision and picked up by a computer-controlled micropipette. This method can be routinely applied as a FACS down to the single cell level with a very high selectivity. Sorting resolution, i.e., the minimum distance between two cells from which one could be selectively removed was 50-70 micrometers. Survival rate with a low number of 3T3 mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was 66 +/- 12% and 88 +/- 16%, respectively. Purity of sorted cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures were 95 +/- 2% and 62 +/- 7%, respectively. Hydrodynamic simulations confirmed the experimental sorting efficiency and a cell damage risk similar to that of normal FACS

DeltaPhish: Detecting Phishing Webpages in Compromised Websites

The large-scale deployment of modern phishing attacks relies on the automatic exploitation of vulnerable websites in the wild, to maximize profit while hindering attack traceability, detection and blacklisting. To the best of our knowledge, this is the first work that specifically leverages this adversarial behavior for detection purposes. We show that phishing webpages can be accurately detected by highlighting HTML code and visual differences with respect to other (legitimate) pages hosted within a compromised website. Our system, named DeltaPhish, can be installed as part of a web application firewall, to detect the presence of anomalous content on a website after compromise, and eventually prevent access to it. DeltaPhish is also robust against adversarial attempts in which the HTML code of the phishing page is carefully manipulated to evade detection. We empirically evaluate it on more than 5,500 webpages collected in the wild from compromised websites, showing that it is capable of detecting more than 99% of phishing webpages, while only misclassifying less than 1% of legitimate pages. We further show that the detection rate remains higher than 70% even under very sophisticated attacks carefully designed to evade our system.Comment: Preprint version of the work accepted at ESORICS 201

Nonlinear 3D Model of Double Shear Lap Tests for the Bond of Near-surface Mounted FRP Rods in Concrete Considering Different Embedment Depth

The utilization of near-surface mounted Fiber Reinforced Polymer (FRP) reinforcement as a method of strengthening in reinforced concrete structures has increased considerably in recent years. Moreover, the application of double-shear lap tests for this rein-forcement method leads to the achievement of a local bond-slip behavior in a bonded joint. This research, therefore, focused on 3-D modeling of this type of test to suitably characterize the bond mechanics between FRP rods and concrete at various embedment depth. The use of different alternatives to represent the interface between the FRP rod and concrete were analyzed after which a&nbsp;comparison was drawn between the numerical finite element (FE) simulations and experimental measurements. The results showed the prediction of the load–slip corresponded with the data obtained from the experiment. Finally, the proposed model has the ability to express the relationship between the penalty stiffness parameters in shear direction Kss = (Ktt) and the embedment depth of FRP rods