Developing a Program Costs Checklist of Digital Health Interventions: A Scoping Review and Empirical Case Study.

INTRODUCTION: The rate of development and complexity of digital health interventions (DHIs) in recent years has to some extent outpaced the methodological development in economic evaluation and costing. Particularly, the choice of cost components included in intervention or program costs of DHIs have received scant attention. The aim of this study was to build a literature-informed checklist of program cost components of DHIs. The checklist was next tested by applying it to an empirical case, Mamma Mia, a DHI developed to prevent perinatal depression. METHOD: A scoping review with a structured literature search identified peer-reviewed literature from 2010 to 2022 that offers guidance on program costs of DHIs. Relevant guidance was summarized and extracted elements were organized into categories of main cost components and their associated activities following the standard three-step approach, that is, activities, resource use and unit costs. RESULTS: Of the 3448 records reviewed, 12 studies met the criteria for data extraction. The main cost categories identified were development, research, maintenance, implementation and health personnel involvement (HPI). Costs are largely considered to be context-specific, may decrease as the DHI matures and vary with number of users. The five categories and their associated activities constitute the checklist. This was applied to estimate program costs per user for Mamma Mia Self-Guided and Blended, the latter including additional guidance from public health nurses during standard maternal check-ups. Excluding research, the program cost per mother was more than double for Blended compared with Self-Guided (€140.5 versus €56.6, 2022 Euros) due to increased implementation and HPI costs. Including research increased the program costs to €190.8 and €106.9, respectively. One-way sensitivity analyses showed sensitivity to changes in number of users, lifespan of the app, salaries and license fee. CONCLUSION: The checklist can help increase transparency of cost calculation and improve future comparison across studies.

Estimating program or intervention costs of digital health interventions (DHIs) can be challenging without a checklist. We reviewed scientific literature to identify key cost categories of DHIs: development, research, maintenance, implementation and health personnel involvement. We also summarized relevant information regarding resource use and unit cost for each of the aforementioned categories. Applying this checklist to Mamma Mia, a DHI aimed at preventing perinatal depression, we find that the total cost per user for Mamma Mia Self-Guided is €106.9 and for Mamma Mia Blended is €190.8. Our checklist enhances visibility of DHI cost components and can aid analysts in better estimating costs for economic evaluations.

Simultaneous RNA and DNA Adductomics Using Single Data-Independent Acquisition Mass Spectrometry Analysis.

Adductomics studies are used for the detection and characterization of various chemical modifications (adducts) of nucleic acids and proteins. The advancements in liquid chromatography coupled with high-resolution tandem mass spectrometry (HRMS/MS) have resulted in efficient methods for qualitative and quantitative adductomics. We developed an HRMS-based method for the simultaneous analysis of RNA and DNA adducts in a single run and demonstrated its application using Baltic amphipods, useful sentinels of environmental disturbances, as test organisms. The novelty of this method is screening for RNA and DNA adducts by a single injection on an Orbitrap HRMS instrument using full scan and data-independent acquisition. The MS raw files were processed with an open-source program, nLossFinder, to identify and distinguish RNA and DNA adducts based on the characteristic neutral loss of ribonucleosides and 2'-deoxyribonucleosides, respectively. In the amphipods, in addition to the nearly 150 putative DNA adducts characterized earlier, we detected 60 putative RNA adducts. For the structural identification of the detected RNA adducts, the MODOMICS database was used. The identified RNA adducts included simple mono- and dimethylation and other larger functional groups on different ribonucleosides and deaminated product inosine. However, 54 of these RNA adducts are not yet structurally identified, and further work on their characterization may uncover new layers of information related to the transcriptome and help understand their biological significance. Considering the susceptibility of nucleic acids to environmental factors, including pollutants, the developed multi-adductomics methodology with further advancement has the potential to provide biomarkers for diagnostics of pollution effects in biota.

Manjari Medika Grape Seed Extract Protects Methotrexate-Induced Hepatic Inflammation: Involvement of NF-κB/NLRP3 and Nrf2/HO-1 Signaling System.

Objective: Grape Seed Extract is a natural source of various polyphenols, which have been shown to possess potent antioxidant and free radical-scavenging activities. The earlier studies have reported that grape seed extract exhibits broad-spectrum pharmacological activities. Therefore, studying the hepatoprotective effects and elucidation of mechanisms of action of the Indian Variety, Manjari Medika grape seed extract (GSE), may give an insight into therapeutic benefits. Methotrexate (MTX) is the first-line pharmacological therapy for different rheumatic diseases. The major adverse events such as hepatotoxicity are evident even in the low doses used for the treatment. The present study investigated the role of MTX on hepatic damage in murine liver and the plausible protective effects of the Indian grape variety, Manjari Medika grape seed extract, in ameliorating it. Methods and Results: To assess the hepatological modulation, mice were divided into eight groups to investigate the ameliorative potential of this GSE (75 and 125 mg/kg) and correlate the experimental findings. The active components of the extract were assessed through UPLC-(ESI)-QToF-MS analysis. On the other hand, various biochemical and immunological indices were carried out to correlate the experimental data. The result demonstrated that the prophylactic administration of GSE reduced MTX-induced hepatic toxicity indices, which subsequently restored the hepatic morphological architecture. Moreover, the application of GSE in a dual dosage (75 and 125 mg/kg) suppressed MTX-induced reactive oxygen species generation, followed by lipid peroxidation and cellular nitrite formation. MTX-induced inflammasome activation through the redox-assisted cascade of TLR4/NF-κB signaling was further reduced by applying the GSE. The results showed that the activation of cytoprotective transcription factor Nrf2 enhanced the level of endogenous antioxidants. Furthermore, through the regulation of TLR4/NF-κB and Nrf2/HO-1 axis, this extract could reduce the MTX-mediated hepatic damage. Conclusion: Our findings suggest that Manjari Medika seed extract could be used as a therapeutic agent to relieve the side effects of MTX and other hepatic disorders.

Heat transfer analysis: convective-radiative moving exponential porous fins with internal heat generation.

The efficiency, temperature distribution, and temperature at the tip of straight rectangular, growing and decaying moving exponential fins are investigated in this article. The influence of internal heat generation, surface and surrounding temperatures, convection-conduction, Peclet number and radiation-conduction is studied numerically on the efficiency, temperature profile, and temperature at the tip of the fin. Differential transform method is used to investigate the problem. It is revealed that thermal and thermo-geometric characteristics have a significant impact on the performance, temperature distribution, and temperature of the fin's tip.The results show that the temperature distribution of decaying exponential and rectangular fins is approximately 15 and 7% higher than growing exponential and rectangular fins respectively. It is estimated that the temperature distribution of the fin increases by approximately 6% when the porosity parameter is increased from 0.1 to 0.5. It is also observed that the decay exponential fin has better efficiency compared to growing exponential fin which offers significant advantages in mechanical engineering.

Genetic Evidence in Favor of a Polyketide Origin of Acremeremophilanes, the Fungal "Sesquiterpene" Metabolites.

Eremophilanes are a large group of "sesquiterpenes" produced by plants and fungi, with more than 180 compounds being known in fungi alone. Many of these compounds are phytotoxic, antimicrobial, anticancer and immunomodulators, and hence are of great economic values. Acremeremophilanes A to O have earlier been reported in a marine isolate of Acremonium sp. We report here the presence of Acremeremophilane I, G, K, N, and O, in a plant beneficial fungus Trichoderma virens, in a strain-specific manner. We also describe a novel, P strain-specific polyketide synthase (PKS) gene cluster in T. virens. This gene cluster, designated amm cluster, is absent in the genome of a Q strain of T. virens, and in other Trichoderma spp.; instead, a near identical cluster is present in the genome of the toxic mold Stachybotrys chartarum. Using gene knockout, we provide evidence that acremeremophilanes are biosynthesized via a polyketide route, and not via the mevalonate/terpene synthesis route as believed. We propose here that the 10-carbon skeleton is a product of polyketide synthase, to which a five-carbon isoprene unit is added by a prenyl transferase (PT), a gene for which is present next to the PKS gene in the genome. Based on this evidence, we propose that at least some of the eremophilanes classified in literature as sesquiterpenes (catalyzed by terpene cyclase) are actually meroterpenes (catalyzed by PKSs and PTs), and that the core moiety is not a sesquiterpene, but a hybrid polyketide/isoprene unit. IMPORTANCE The article contradicts the established fact that acremeremophilane metabolites produced by fungi are sesquiterpenes; instead, our findings suggest that at least some of these well-studied metabolites are of polyketide origin. Acremeremophilane metabolites are of medicinal significance, and the present findings have implications for the metabolic engineering of these metabolites and also their overproduction in microbial cell factories.

Coupling the high-resolution LC-MS characterisation of the phenolic compounds with the antimicrobial and antibiofilm properties of helencha (Enydra fluctuans Lour.).

This study reports the polyphenol profile of helencha (Enydra fluctuans Lour.), an underutilised, aquatic leafy vegetable, based on high resolution accurate mass analysis. The methanolic extract of helencha leaves was screened by ultra-high performance liquid chromatography with quadrupole time of flight mass spectrometry (LC-QToF-MS). An in-house developed database of phytochemical metabolites was referred for compound identifications. Based on the detection of the pseudomolecular ion and at least one molecule-specific fragment ion (each with < 5 ppm of mass error), 25 potentially-bioactive phenolic compounds were putatively identified. These included 6 flavonols, 4 phenolic acids, 3 lignans, 3 flavones and 1 each of flavanol, flavanone, dihydroflavonol, tetramethoxyflavone, isoflavonoid and methylated flavonol. In addition, 3 unclassified compounds are also reported. The helencha extract showed antibiofilm properties with a potent bacteriostatic activity against the clinical isolates of Pseudomonas aeruginosa, a human pathogenic bacteria. The complementary molecular docking studies indicated strong binding interactions of the identified compounds with the active site of LasR protein of P. aeruginosa. The in vitro and in silico study results would be useful to develop novel neutraceutical products based on helencha-extract and design new lead compounds to control the biofilm producing pathogenic microorganisms. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-021-04968-y).

Dual role of a dedicated GAPDH in the biosynthesis of volatile and non-volatile metabolites- novel insights into the regulation of secondary metabolism in Trichoderma virens.

Trichoderma virens produces viridin/viridiol, heptelidic (koningic) acid, several volatile sesquiterpenes and gliotoxin (Q strains) or gliovirin (P strains). We earlier reported that deletion of the terpene cyclase vir4 and a glyceraldehyde-3-phosphate dehydrogenase (GAPDH, designated as vGPD) associated with the "vir" cluster abrogated the biosynthesis of several volatile sesquiterpene metabolites. Here we show that, the deletion of this GAPDH also impairs the biosynthesis of heptelidic acid (a non-volatile sesquiterpene), viridin (steroid) and gliovirin (non-ribosomal peptide), indicating regulation of non-volatile metabolite biosynthesis by this GAPDH that is associated with a secondary metabolism gene cluster. To gain further insights into the details of this novel form of regulation, we identified the terpene cyclase gene responsible for heptelidic acid biosynthesis (hereafter designated as has1) and prove that the expression of this gene is regulated by vGPD. Interestingly, deletion of has1 impaired biosynthesis of heptelidic acid (HA), viridin and gliovirin, but not of volatile sesquiterpenes. Deletion of the vir cluster associated terpene cyclase gene (vir4), located next to the vGPD gene, did not impair biosynthesis of HA, viridin or gliovirin. We thus unveil a novel circuitry of regulation of secondary metabolism where an HA-tolerant GAPDH isoform (vGPD) regulates HA biosynthesis through the transcriptional regulation of the HA-synthase gene (which is not part of the "vir" cluster). Interestingly, impairment of HA biosynthesis leads to the down-regulation of biosynthesis of other non-volatile secondary metabolites, but not of volatile secondary metabolites. We thus provide evidence that the "vir" cluster associated, HA-tolerant GAPDH in T. virens participates in the biosynthesis of volatile sesquiterpenes as a biosynthetic enzyme, and regulates the production of non-volatile metabolites via regulation of HA biosynthesis. The orthologue of the "vir" cluster in Aspergillus oryzae was earlier reported to synthesize HA by another group. Our study thus proves that the same gene cluster can code for unrelated metabolites in different species.

Multiclass pesticide residue analysis in tobacco (Nicotiana tabacum) using high performance liquid chromatography-high resolution (Orbitrap) mass spectrometry: A simultaneous screening and quantitative method.

Public exposure to pesticides through tobacco has attracted serious attention. Here we report a simultaneous screening and quantitation method for the non-target multiresidue analysis of pesticides in different tobacco types. The method involved extraction of a homogenate (20 g, containing 2 g tobacco) in ethyl acetate (10 mL), cleanup of 2 mL extract by dispersive solid phase extraction with PSA (50 mg)+C18 (50 mg)+GCB (25 mg)+MgSO4 (100 mg), followed by reconstitution in 1 mL acetonitrile:water (3:7) and analysis using HPLC with Quadrupole-Orbitrap mass spectrometry. The high resolution accurate mass analysis was performed through sequential full-scan (resolution=35000) and variable data independent acquisition (resolution=17500) events. When the method was evaluated in a mixture of 181 pesticides, it effectively minimised matrix interferences and false negatives. The target compounds included 5 pairs of isomers and 27 pairs of isobars, which were distinguished based on chromatographic separation, mass resolving power and/or unique product ions. The screening detection limit (SDL) for 86.4% of the test pesticides was set at 5 ng/g, while the remainder had the SDLs at 10 ng/g (9.3%) and 40 ng/g (4.3%). Nearly, 75% of the compounds showed recoveries of 70-120% at 10 ng/g. The rest of the compounds showed satisfactory recoveries at 40 and 100 ng/g. In all cases, precision-RSDs were < 20%. The established method demonstrated a successful performance in four different types of tobacco matrices while aligning with the guidelines of SANTE and US-FDA. Owing to its efficiency, the method is recommended for screening and quantitation of multiclass pesticides in tobacco.

Antityrosinase Activity of Combretum micranthum, Euphorbia hirta and Anacardium occidentale Plants: Ultrasound Assisted Extraction Optimization and Profiling of Associated Predominant Metabolites.

Tyrosinase is an important component of the enzyme polyphenol oxidase, which upon contact with the phenolic substrates forms the pigment melanin and induces undesirable food browning. The phenolic and triterpenoid compounds that naturally occur in plants are well known as tyrosinase inhibitors. Combretum micranthum (CM) leaves, Euphorbia hirta (EH) plant, and Anacardium occidentale (AO) fruits are traditionally known to have potential anti-tyrosinase activities. The aim of this study was to optimize the ultrasound-assisted extraction of secondary metabolites from these matrices, and to evaluate in tubo the antityrosinase activity of these extracts. Efforts were also taken to profile the secondary metabolites, mainly the phenolic and triterpenoid compounds, in order to understand their probable association with tyrosinase inhibition. The optimal ultrasound-assisted extraction conditions for simultaneous extraction of phenolic, and triterpenoid compounds were determined. The aqueous fraction of these extracts showed significant antityrosinase activity, with the CM leaves exhibiting the strongest inhibitory effect (IC50 of 0.58 g·L-1). The predominant metabolic compounds from these natural extracts were putatively identified by using a high-resolution quadrupole-time of flight (QToF) LC-MS instrument. The high-resolution accurate mass-based screening resulted in identification of 88 predominant metabolites, which included dihydrodaidzein-7-O-glucuronide, micromeric acid, syringic acid, morin, quercetin-3-O-(6″-malonyl-glucoside), 4-hydroxycoumarin, dihydrocaffeic acid-3-O-glucuronide, to name some, with less than 5 ppm of mass error.

Profiling of polyphenols in phalsa (Grewia asiatica L) fruits based on liquid chromatography high resolution mass spectrometry.

The aim of the study was to screen the metabolite profile of phalsa (Grewia asiatica), an underutilized fruit crop, using liquid chromatography-high resolution mass spectrometric analysis. A total of 50 compounds were tentatively identified based on their molecular mass and characteristic fragment ions, each with less than 5 ppm of mass error. These compounds included 21 flavonols, 2 dihydroflavonols, 7 flavones, 3 flavanols, 6 anthocyanins, 3 isoflavonoids, 2 phenolic acids, 2 flavanones, and 4 other phenolics. Flavonols were the predominant group of compounds, representing around 52.6% of the total phenolics. The paper has also discussed the potentiality of phalsa as an emerging functional food for the management of various human diseases in relation to the existing literature.

Anthocyanin Profiling Using UV-Vis Spectroscopy and Liquid Chromatography Mass Spectrometry.

BACKGROUND: As a powerful antioxidant and natural colorant, anthocyanins are being used increasingly as a component of food supplements and nutraceutical products. Hence, its characterization is a prerequisite for further exploration of its nutraceutical potential. UV-Vis and MS are the two important techniques, which have been largely employed for the qualitative and quantitative determination of anthocyanins. However, a comprehensive review of the applications of these techniques in literature is scarce. OBJECTIVE: This paper aims to review the utilization of UV-Vis spectral data as well as mass spectral data for characterization and putative identification of anthocyanins with approaches of quantification. METHODS: The techniques described in literature have been thoroughly reviewed and comparatively evaluated. The complementary approaches of UV-Vis and MS spectra have been discussed for identification and quantification of these compounds. RESULTS: Valuable information about the chemical composition and structure of anthocyanins can be predicted from the UV-Vis spectral data, such as number and type of glycosylation as well as absence or presence of acylation, to name a few. It is also pointed out that for their structural confirmation, selectivity of mass detectors with unit and high-resolution analysis could be effective. CONCLUSIONS: The combination of LC-MS with UV-Vis spectroscopy provides complementary information on structural details of anthocyanins. In case the analytical reference standards are available, a triple quadrupole mass spectrometer provides selectivity and quantitative sensitivity in analysis. On the other hand, high-resolution MS analysis provides valuable information for tentative identification during nontarget screening of compounds when the reference standard is not available. HIGHLIGHTS: This paper reviews the applications of UV-Vis spectroscopy and LC-MS for qualitative and quantitative analysis of anthocyanins.

Elucidation of False Detection of Pesticides During Residue Analysis in Indian Tobacco by Multidimensional GC-MS.

BACKGROUND: False detection of pesticides in agricultural produce may raise serious questions regarding both consumer safety and trade. High levels of delta-hexachlorocyclohexane (δ-HCH; 11.7-22.3 mg/kg) were detected in some tobacco samples in a retention time-based GC analysis. Hence, the selection of an appropriate analytical method is an uncompromisable necessity. OBJECTIVES: This research work aimed to elucidate false detection of pesticides along with identification of coeluting tobacco matrix compounds to understand the dynamics of false detection with an increase in the number of analyzed pesticides and to screen suitable analytical methods. METHODS: Initially, retention time-based GC analysis was performed for monitoring of 10 pesticide residues in tobacco leaf matrix, followed by GC-MS/selected-ion monitoring (SIM) analysis. Then, the total number of pesticides to be analyzed was increased to 47, and residue analysis was performed by involving GC-MS/SIM and multidimensional (MD) GC-MS. RESULTS: A false-positive detection of δ-HCH due to a coeluting tobacco aroma compound, neophytadiene, during residue analysis of 10 pesticides in tobacco (Nicotiana tabacum L.) leaf was observed. This problem was resolved by employing the unique quantifier and qualifier ions in SIM mode. However, with 47 pesticides, neophytadiene completely masked the signal of δ-HCH, which resulted in an impure spectrum of δ-HCH (<30% similarity match) even after application of selective quantifier and qualifier ions. Finally, MDGC-MS analysis could resolve it by chromatographic separation of the said analyte from the coeluting matrix compound. CONCLUSIONS: The findings of this work offer the potential to minimize false reporting of target pesticides to comply with consumer safety and trade standards. HIGHLIGHTS: The study identifies various tobacco matrix compounds coeluting with pesticides during multiresidue analysis. Neophytadiene, a tobacco aroma compound, resulted in false-positive detection of δ-HCH. The MDGC-MS could be effectively used as a confirmatory analysis tool for reliable detection of pesticide residue in tobacco leaf matrix.

Improved analysis of captan, tetrahydrophthalimide, captafol, folpet, phthalimide, and iprodione in fruits and vegetables by liquid chromatography tandem mass spectrometry.

An improved liquid chromatography tandem mass spectrometry method is reported for the determination of residues of captan (+tetrahydrophthalimide), captafol, folpet (+phthalimide), and iprodione in fruits and vegetables. The optimized electrospray ionization parameters (high cone gas flow, and a low desolvation temperature) did not result in degradation of target compounds, rather they provided a significant advantage over the conventional GC-MS/MS methods, which lack sensitivity and repeatability. Strategies for minimizing losses in recovery of these compounds during sample preparation included cryogenic comminution, extraction with acidified ethyl acetate or acetonitrile, and dilution of the final extract with acidified water prior to LC-MS/MS analysis. The method performance complied with the SANTE/11813/2017 guidelines, with recoveries in the range of 70-120% at the LOQ of 0.01 mg/kg across the tested matrices at various pHs. The efficiency of the method was reflected in its precision (RSDs < 10%) for incurred residues.

Targeted phenolic profiling of Sauvignon blanc and Shiraz grapes grown in two regions of India by liquid chromatography-tandem mass spectrometry.

The phenolic compounds play an important role in production of quality grapes and wines. The current investigation focused on optimization of an extraction method for targeted analysis of 33 phenolic compounds in grapes by liquid chromatography tandem mass spectrometry (LC-MS/MS). The optimized method was successfully used for phenolic profiling of two wine grape varieties, Sauvignon blanc (white) and Shiraz (red) originated from Pune and Nasik regions of Maharashtra State, India. The optimized sample preparation procedure involved liquid-liquid extraction with acidified methanol by vortexing for 2 min followed by analysis on LC-MS/MS. The limit of quantification of the targeted compounds was in the range of 29 to 411 µg/L. The results indicated that skin of both varieties contained the highest amount of flavonols (69.47 ± 14.74 mg/kg in Sauvignon blanc and 129.47 ± 10.05 mg/kg in Shiraz) compared to pulp. The highest amounts of flavan-3-ols were present in grape seed collected from the Pune region (2016.84 ± 14.73 mg/kg in Sauvignon blanc and 1945.06 ± 32.69 mg/kg in Shiraz). The concentration of stilbenes was the highest in grape skin (0.13 ± 0.52 to 5.78 ± 5.45 mg/kg) compared to seed and pulp of both varities. Hydroxybenzoic acid (vanillin), hydroxycinnamic acid (p-coumaric acid) and anthocyanins (oenin, malvidin, cyanidin and kuromanin) were found only in Shiraz variety. The results of antioxidant activity (FRAP and DPPH assay) indicated the highest scavenging activity in seed (978.64 ± 56.23 to1133.38 ± 143.65 µMol TE/g DW FRAP and 594.93 ± 37.94 to 631.94 ± 56.45 µMol TE/g DW in DPPH). The phenolic contents in Sauvignon blanc and Shiraz grapes between Pune and Nasik regions did not have any significant difference.

The Effect of Microbial Endophyte Consortia on Pseudotsuga menziesii and Thuja plicata Survival, Growth, and Physiology Across Edaphic Gradients.

Increased frequency of droughts and degraded edaphic conditions decreases the success of many reforestation efforts in the Pacific Northwest. Microbial endophyte consortia have been demonstrated to contribute to plant growth promotion and protection from abiotic and biotic stresses - specifically drought conditions - across a number of food crops but for limited tree species. Our research aimed to investigate the potential to improve establishment of economically and ecologically important conifers through a series of in situ field trials and ex situ simulations. Microbial endophyte consortia from Salicaceae, previously shown to confer drought tolerance, and conifer endophyte strains with potentially symbiotic traits were selected for trials with Douglas-fir (Pseudotsuga menziesii) and western redcedar (Thuja plicata). Reductive experimentation was used to subject seedlings to a spectrum of simulated drought levels or presence/absence of fertilizer, testing hypotheses that endophyte consortia impart improved drought resistance and growth promotion, respectively. Inoculation from Salicaceae consortia significantly (p ≤ 0.05) improved survival among seedlings of both species subject to increasing drought stress, with T. plicata seedlings surviving at twofold higher rates in extreme drought conditions. Both species demonstrated improved growth 540 days after inoculation of seed with conifer derived consortia. In the carefully controlled greenhouse experiments with both species, seedling Fv/Fm and SPAD values remained significantly (p ≤ 0.05) more stable in inoculated treatment groups as stress increased. Our findings confirm that multi-strain consortia may be applied as seed or field amendment to conifers, and the approach is efficient in garnering a positive growth response and can mitigate abiotic stressors.

High throughput residue analysis of paraquat and diquat involving hydrophilic interaction liquid chromatographic separation and mass spectrometric determination.

A selective, sensitive and robust LC-MS/MS method is reported for the determination of the residues of paraquat and diquat in various fruit matrices, including grape, apple and pomegranate. The extraction with acidified water (0.1 M HCl) at 80°C (15 min) offered superior recoveries for both analytes with a significantly lower matrix effects as compared to the extraction with acidified methanol by the methods reported in the existing literature. The optimised HPLC conditions on hydrophilic interaction liquid chromatography (HILIC) columns, when coupled with electrospray ionisation-tandem mass spectrometry, offered their limit of quantification at 0.01 mg kg-1. The analysis on an XBridge HILIC column required a thorough optimisation of the gradient programme to induce chromatographic separation and minimise matrix effects. This was not necessary when a CORTECS HILIC column was used, which provided selective and sensitive analysis within 5 min runtime using isocratic flow. Isotopically labelled internal standards corrected the recoveries of both analytes within 70-120% (RSD < 20%). For the first time, the applications of high resolution accurate mass analysis in the 'time of flight - multiple reaction monitoring' mode have been demonstrated as a complementary means of targeted screening of these compounds at 0.01 mg kg-1 level. The method has a strong potential for applications in both official control and by those involved in food production for checking compliance with the EU MRLs.

Comprehensive multiresidue determination of pesticides and plant growth regulators in grapevine leaves using liquid- and gas chromatography with tandem mass spectrometry.

Grape leaf, which is known for its nutritional and medicinal properties, is finding increased applications for cuisine and remedial purposes. This article reports a comprehensive analytical method for the identification and quantification of a broad range of pesticides and plant growth regulators (PGRs) in the grape leaf matrix. The sample preparation method for pesticides involved an optimized QuEChERS-based extraction protocol, with subsequent clean-up by the dispersive solid phase extraction (dSPE) using a mixture of sorbents (25 mg PSA + 5 mg GCB + 150 mg MgSO4). The PGRs were extracted with methanol. The performance of the method was investigated and validated for a mixture of 363 pesticides (148 in GC-MS/MS and 203 in LC-MS/MS) and 12 PGRs (LC-MS/MS) in compliance with the analytical quality control criteria of the SANTE/11813/2017 guidelines. The matrix effects were comparatively higher against grape berries. The findings indicated satisfactory recoveries at 10 ng/g and higher levels with precision RSDs less than 20%. This method has potential applications in commercial residue testing laboratories and also for the regulatory compliance check purposes for its lower LOQs compared to the corresponding EU-MRLs.

Analysis of pesticide residues in tuber crops using pressurised liquid extraction and gas chromatography-tandem mass spectrometry.

Tuber crops substantially contribute to the food security in the developing countries. Often, their cultivation involves unregulated applications of pesticides, leading to MRL non-compliances. Despite their rising currency in international trade, there exist scarcely any methods for pesticide residue analysis in these matrices. Therefore, we developed a multi-residue method for simultaneous analysis of a diverse range of pesticides in tuber crops, based on pressurized liquid extraction by ethyl acetate, followed by selective identification and quantification of the residues using GC-MS selected reaction monitoring. The method was evaluated for 150 pesticides. Results showed that their limits of quantification were 0.1-10ng/g, with recoveries of 70-120%. When compared to the conventional analytical techniques, such as QuEChERS and buffered ethyl acetate extraction, this method provided superior performance in terms of precision, and recovery of the spiked and incurred residues with similar productivity. The method holds promise for commercial and regulatory residue analysis.

High-sensitivity direct analysis of aflatoxins in peanuts and cereal matrices by ultra-performance liquid chromatography with fluorescence detection involving a large volume flow cell.

This paper reports a sensitive and cost effective method of analysis for aflatoxins B1, B2, G1 and G2. The sample preparation method was primarily optimised in peanuts, followed by its validation in a range of peanut-processed products and cereal (rice, corn, millets) matrices. Peanut slurry [12.5 g peanut + 12.5 mL water] was extracted with methanol: water (8:2, 100 mL), cleaned through an immunoaffinity column and thereafter measured directly by ultra-performance liquid chromatography-fluorescence (UPLC-FLD) detection, within a chromatographic runtime of 5 minutes. The use of a large volume flow cell in the FLD nullified the requirement of any post-column derivatisation and provided the lowest ever reported limits of quantification of 0.025 for B1 and G1 and 0.01 µg/kg for B2 and G2. The single laboratory validation of the method provided acceptable selectivity, linearity, recovery and precision for reliable quantifications in all the test matrices as well as demonstrated compliance with the EC 401/2006 guidelines for analytical quality control of aflatoxins in foodstuffs.

A simultaneous screening and quantitative method for the multiresidue analysis of pesticides in spices using ultra-high performance liquid chromatography-high resolution (Orbitrap) mass spectrometry.

A novel screening and quantitation method is reported for non-target multiresidue analysis of pesticides using ultra-HPLC-quadrupole-Orbitrap mass spectrometry in spice matrices, including black pepper, cardamom, chili, coriander, cumin, and turmeric. The method involved sequential full-scan (resolution = 70,000), and variable data independent acquisition (vDIA) with nine consecutive fragmentation events (resolution = 17,500). Samples were extracted by the QuEChERS method. The introduction of an SPE-based clean-up step through hydrophilic-lipophilic-balance (HLB) cartridges proved advantageous in minimizing the false negatives. For coriander, cumin, chili, and cardamom, the screening detection limit was largely at 2 ng/g, while it was 5 ng/g for black pepper, and turmeric. When the method was quantitatively validated for 199 pesticides, the limit of quantification (LOQ) was mostly at 10 ng/g (excluding black pepper, and turmeric with LOQ = 20 ng/g) with recoveries within 70-120%, and precision-RSDs <20%. Furthermore, the method allowed the identification of suspected non-target analytes through retrospective search of the accurate mass of the compound-specific precursor and product ions. Compared to LC-MS/MS, the quantitative performance of this Orbitrap-MS method had agreements in residue values between 78-100%.