Food Safety Testing Research Articles

Metagenomic analysis of medicinal Cannabis samples; pathogenic bacteria, toxigenic fungi, and beneficial microbes grow in culture-based yeast and mold tests.

Background: The presence of bacteria and fungi in medicinal or recreational Cannabis poses a potential threat to consumers if those microbes include pathogenic or toxigenic species. This study evaluated two widely used culture-based platforms for total yeast and mold (TYM) testing marketed by 3M Corporation and Biomérieux, in comparison with a quantitative PCR (qPCR) approach marketed by Medicinal Genomics Corporation. Methods: A set of 15 medicinal Cannabis samples were analyzed using 3M and Biomérieux culture-based platforms and by qPCR to quantify microbial DNA. All samples were then subjected to next-generation sequencing and metagenomics analysis to enumerate the bacteria and fungi present before and after growth on culture-based media. Results: Several pathogenic or toxigenic bacterial and fungal species were identified in proportions of >5% of classified reads on the samples, including Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Ralstonia pickettii, Salmonella enterica, Stenotrophomonas maltophilia, Aspergillus ostianus, Aspergillus sydowii, Penicillium citrinum and Penicillium steckii. Samples subjected to culture showed substantial shifts in the number and diversity of species present, including the failure of Aspergillus species to grow well on either platform. Substantial growth of Clostridium botulinum and other bacteria were frequently observed on one or both of the culture-based TYM platforms. The presence of plant growth promoting (beneficial) fungal species further influenced the differential growth of species in the microbiome of each sample. Conclusions: These findings have important implications for the Cannabis and food safety testing industries.

The Application and Development of Measurement in Guaranteeing the Food Safety in China

Measurement plays an important role in the process of food safety testing. The study of the application and development of measurement in guaranteeing food safety is of great theoretical and guiding significance. This study expounds the concept of measurement and the recent application of measurement in guaranteeing the food safety in the domestic and international areas and analyzes the function of measurement in the detection of food safety. Furthermore, some useful countermeasures and suggestions are put forward to improve the measurement system of food in China. The aim of study is to provide a reference for establishing a perfect mechanism for the implementation of food measuring work.

Dual FITC lateral flow immunoassay for sensitive detection of Escherichia coli O157:H7 in food samples

A pattern of signal amplification lateral flow immunoassay (LFIA) for pathogen detection, which used fluorescein isothiocyanate (FITC) labeled antigen and antibody for dual FITC-LFIA was developed. Escherichia coli O157:H7 (E.coli O157:H7) was selected as the model analyte. In the signal amplification LFIA method, FITC was mixed with sample culture medium, with the presence of E.coli O157:H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation, creating a fluorescent antigen probe. This antigen probe was added to LFIA, which already contained E.coli O157:H7 monoclonal antibodies-FITC (McAb-E.coli O157:H7-FITC) dispersed in the conjugate pad. Another E.coli O157:H7 McAb was the test line, and goat anti-mouse IgG antibody was the control line in nitrocellulose (NC) membrane. The visual limit of detection (LOD) of the strip for qualitative detection was 105 CFU/mL while the LOD for semi-quantitative detection could down to 104 CFU/mL by using scanning reader. Signal amplification LFIA was perfectly applied to the detection of food samples with E.coli O157:H7. The LOD was substantially improved to 1 CFU/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 8 and 8h respectively. The results of this method was more sensitive by 10-fold than the conventional colloidal gold (CG) based strips and comparable to the traditional ELISA. This simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring food-borne pathogens in food-safety testing.

Cannabis microbiome sequencing reveals several mycotoxic fungi native to dispensary grade Cannabis flowers.

The Center for Disease Control estimates 128,000 people in the U.S. are hospitalized annually due to food borne illnesses. This has created a demand for food safety testing targeting the detection of pathogenic mold and bacteria on agricultural products.This risk extends to medical Cannabis and is of particular concern with inhaled, vaporized and even concentrated Cannabis products . As a result, third party microbial testing has become a regulatory requirement in the medical and recreational Cannabis markets, yet knowledge of the Cannabis microbiome is limited. Here we describe the first next generation sequencing survey of the fungal communities found in dispensary based Cannabis flowers by ITS2 sequencing, and demonstrate the sensitive detection of several toxigenic Penicillium and Aspergillus species, including P. citrinum and P. paxilli, that were not detected by one or more culture-based methods currently in use for safety testing.

Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor

A simple, one-step, rapid method to detect Escherichia coli O157: H7 (E. coli O157: H7) using a label-free immunofluorescence strip sensor is presented. Fluorescein isothiocyanate (FITC) was added to the sample culture medium to prepare the fluorescent probe for the label-free strip sensor. With the presence of E. coli O157: H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation and maintain good affinity to the monoclonal antibodies (McAb) against E. coli O157: H7. The direct-type immunofluorescence strip sensor was based on the binding between fluorescent bacteria and the unlabeled McAb immobilized at the test line in nitrocellulose membrane (NC membrane) reaction zone. The visual limit of detection (LOD) of the strip for qualitative detection was 106cells/mL while the LOD for semi-quantitative detection could go down to 105cells/mL by using scanning reader. The LOD was substantially improved to 1cells/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 10 and 8h respectively, which was competitive to some current rapid E. coli O157: H7 detection methods. Besides the obvious advantages, including reduced detection time and operation procedures, the results of this method meet the various detection requirements for E. coli O157: H7 and are comparable to the traditional enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich gold-labeled strips. This is the first report of semi-quantitative immunofluorescence strip for directly detecting foodborne pathogen using only one unlabeled antibody. All detections could be achieved in less than 5min. In addition, this simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring foodborne pathogens in food-safety testing.

Efficient Capture of Particles Via Rotating Magnetic Beads in a Microfluidic Channel

Even with stringent levels of detection the CDC estimates 48 million people a year suffer from food poisoning (1, 2). While it is true that deaths from food poisoning are rare, the economic impact from food poisoning is estimated at $77 billion dollars a year (3). Rapid and efficient detection in food safety applications has two primary challenges. First, the FDA detection level for bacterial contamination is one live cell per 20 grams of food material, a large volume. The practicality of finding one prokaryotic cell in the volume requires pre-concentration as a crucial step in improving the reliability of the test. Second, it is possible to find multiple strains of a given bacteria in a sample. Each of these strains will have similar morphology but different pathological results. Thus a system is required that is strain specific which can also process large quantities of sample. Techniques for cell sorting in microfluidic systems have received a lot of attention recently. The most popular methods to separate the cells are based upon physical characteristics, such as size, shape, weight, density or elasticity. These systems often utilize passive structures that rely on flow conditions or size exclusion principles. However, sorting via this method is not useful for food safety applications because of the strain problem previously described. Labeling the targets with fluorescent or bead-based signals is inefficient due to the large sample volumes involved in food safety testing. This work presents a system that captures dilute concentrations of particles from a bulk fluid (4, 5). The particles are captured by magnetic beads inside a microfluidic channel. The magnetic beads are actuated by rotating around soft magnetic features using an external magnetic field. The magnetic bead sweeps through the fluid, increasing the chances of the particles coming into contact with the magnetic bead by increased fluid mixing and flow agitation (6). Protein and antibody coatings on the magnetic bead then capture the target particles from the bulk fluid and trap it with the magnetic bead. This system is easily multiplexedacross multiple channels to process large quantities of fluid sample and incorporated into a portable system.The system described here uses commercially available magnetic beads (Dynabeads M-280, Life Technologies) and soft magnetic features of permalloy (80/20 NiFe). The soft magnetic features are made by electron beam evaporation onto a bare glass wafer. A standard photolithography process with wet etching of the excess metal was used to produce the soft magnetic features. A layer of silicon dioxide is deposited via plasma enhanced chemical vapor deposition (PECVD) over the entire wafer to protect the soft magnetic features from oxidation during experiments. Finally a PDMS microfluidic channel is created from standard PDMS molding and placed over the wafer to create a fully sealed channel. Magnetic beads are pumped into the array where the magnetic forces from the external magnetic field and the soft magnetic features distribute the beads across the entire array. In this work fluorescent particles (Fluorspheres, 1 µm diameter) are pumped through the array of rotating magnetic beads at various conditions and images taken of the fluorescent particles capture in the array. These particles are analogous in size to common bacteria sought in food safety testing. The goal is to quantify the effectiveness of this system to capture targets from the bulk fluid under different flow and geometric conditions. The conditions considered in this work are flow rate, bead rotation speed, the ratio of magnetic bead diameter to channel height and the packing density of the magnetic features in the array.

Cannabis microbiome sequencing reveals several mycotoxic fungi native to dispensary grade Cannabis flowers

The Center for Disease Control estimates 128,000 people in the U.S. are hospitalized annually due to food borne illnesses. This has created a demand for food safety testing targeting the detection of pathogenic mold and bacteria on agricultural products. This risk extends to medical Cannabis and is of particular concern with inhaled, vaporized and even concentrated Cannabis products . As a result, third party microbial testing has become a regulatory requirement in the medical and recreational Cannabis markets, yet knowledge of the Cannabis microbiome is limited. Here we describe the first next generation sequencing survey of the fungal communities found in dispensary based Cannabis flowers by ITS2 sequencing, and demonstrate the sensitive detection of several toxigenic Penicillium and Aspergillus species, including P. citrinum and P. paxilli, that were not detected by one or more culture-based methods currently in use for safety testing.

Metagenomic analysis of viruses associated with field-grown and retail lettuce identifies human and animal viruses

The emergence of culture- and sequence-independent metagenomic methods has not only provided great insight into the microbial community structure in a wide range of clinical and environmental samples but has also proven to be powerful tools for pathogen detection. Recent studies of the food microbiome have revealed the vast genetic diversity of bacteria associated with fresh produce. However, no work has been done to apply metagenomic methods to tackle viruses associated with fresh produce for addressing food safety. Thus, there is a little knowledge about the presence and diversity of viruses associated with fresh produce from farm-to-fork. To address this knowledge gap, we assessed viruses on commercial romaine and iceberg lettuces in fields and a produce distribution center using a shotgun metagenomic sequencing targeting both RNA and DNA viruses. Commercial lettuce harbors an immense assemblage of viruses that infect a wide range of hosts. As expected, plant pathogenic viruses dominated these communities. Sequences of rotaviruses and picobirnaviruses were also identified in both field-harvest and retail lettuce samples, suggesting an emerging foodborne transmission threat that has yet to be fully recognized. The identification of human and animal viruses in lettuce samples in the field emphasizes the importance of preventing viral contamination on leafy greens starting at the field. Although there are still some inherent experimental and bioinformatics challenges in applying viral metagenomic approaches for food safety testing, this work will facilitate further application of this unprecedented deep sequencing method to food samples.

Cannabis microbiome sequencing reveals several mycotoxic fungi native to dispensary grade Cannabis flowers

The Center for Disease Control estimates 128,000 people in the U.S. are hospitalized annually due to food borne illnesses. This has created a demand for food safety testing targeting the detection of pathogenic mold and bacteria on agricultural products.This risk extends to medical Cannabis and is of particular concern with inhaled, vaporized and even concentrated Cannabis products . As a result, third party microbial testing has become a regulatory requirement in the medical and recreational Cannabis markets, yet knowledge of the Cannabis microbiome is limited. Here we describe the first next generation sequencing survey of the microbial communities found in dispensary based Cannabis flowers and demonstrate the limitations in the culture-based regulations that are being superimposed from the food industry.

A baseline correction algorithm for Raman spectroscopy by adaptive knots B-spline

The Raman spectroscopy technique is a powerful and non-invasive technique for molecular fingerprint detection which has been widely used in many areas, such as food safety, drug safety, and environmental testing. But Raman signals can be easily corrupted by a fluorescent background, therefore we presented a baseline correction algorithm to suppress the fluorescent background in this paper. In this algorithm, the background of the Raman signal was suppressed by fitting a curve called a baseline using a cyclic approximation method. Instead of the traditional polynomial fitting, we used the B-spline as the fitting algorithm due to its advantages of low-order and smoothness, which can avoid under-fitting and over-fitting effectively. In addition, we also presented an automatic adaptive knot generation method to replace traditional uniform knots. This algorithm can obtain the desired performance for most Raman spectra with varying baselines without any user input or preprocessing step. In the simulation, three kinds of fluorescent background lines were introduced to test the effectiveness of the proposed method. We showed that two real Raman spectra (parathion-methyl and colza oil) can be detected and their baselines were also corrected by the proposed method.

EVALUATION OF A LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR THE DETECTION OF VIRUSES IN READY-TO-EAT FOODS

The ability of viruses to persist in the environment and on fresh produce, as well as their low infectious doses, allows even a small amount of contamination to cause foodborne viral outbreaks. Human Adenoviruses (hAdVs) of subgroup F (enteric serotypes 40 and 41) are known to be associated with 5–20% worldwide of acute gastroenteritis cases among infants and young children. The present study aimed to evaluate a simple and cost-effective, one-step, single-tube adenovirus type 40/41 specific loop-mediated isothermal amplification (LAMP) assay for the detection of hAdV40/41 DNA in a series of ready-to-eat food samples (strawberries, sour cherries, lettuce, cherry tomatoes and green onions). No LAMP assay has previously been reported for the detection of these virus types in food samples. Results were obtained within 60 min, under isothermal conditions at 69°C. After amplification, the products were detected either by observing a ladder pattern following gel electrophoresis, or a color change with the addition of SYBR Green I to the reaction tube. The limits of detection of hAdV40/41 in food samples were found to be >30 GCs per reaction, when a nucleic acids extraction step was included in the analysis. A 1 log higher (>100 GCs/reaction) detection limit was found for lettuce, cherry tomatoes and green onions, without a nucleic acids extraction step included before the isothermal amplification. The LAMP assay for the virological analysis of food samples is expected to provide a robust, innovative, powerful, cheap and fast monitoring tool, without the need of sophisticated equipment, which will be available for food safety testing by the food industry and the public food health authorities.

Research of Food Safety Detection Based on Multi-sensor Data Fusion Technology

This study presents a method for food safety testing based on multi-sensor data fusion, mainly to deal with food safety testing to detect the most common substances, these materials include: formaldehyde, heavy metals lead and cadmium and organ phosphorus and carbamate pesticides. Because it is more sensors, so the detection system will generate a lot of signals. Limited ability of the sensor output signals, these signals are often relatively weak and because the system will be applied to on-site testing and site environment is complex signal is easily influenced by many factors. System design related circuit, using a variety of chips, purify and amplify the signal processing of analog to digital conversion. The final article, but also on data fusion technology relevant circumstances were introduced to the relevant algorithm analysis shows. The experiment data shows that the system has a wide detection range and the configuration is simple with low detection cost which shows the method prompted can meet the engineering requirements of food safety detection.

Two-Photon Sensing and Imaging of Endogenous Biological Cyanide in Plant Tissues Using Graphene Quantum Dot/Gold Nanoparticle Conjugate.

One main source of cyanide (CN(-)) exposure for mammals is through the plant consumption, and thus, sensitive and selective CN(-) detection in plants tissue is a significant and urgent work. Although various fluorescence probes have been reported for CN(-) in water and mammalian cells, the detection of endogenous biological CN(-) in plant tissue remains to be explored due to the high background signal and large thickness of plant tissue that hamper the effective application of traditional one-photo excitation. To address these issues, we developed a new two-photo excitation (TPE) nanosensor using graphene quantum dots (GQDs)/gold nanoparticle (AuNPs) conjugate for sensing and imaging endogenous biological CN(-). With the benefit of the high quenching efficiency of AuNPs and excellent two-photon properties of GQDs, our sensing system can achieve a low detection limit of 0.52 μM and deeper penetration depth (about 400 μm) without interference from background signals of a complex biological environment, thus realizing sensing and imaging of CN(-) in different types of plant tissues and even monitoring CN(-) removal in food processing. To the best of our knowledge, this is the first time for fluorescent sensing and imaging of CN(-) in plant tissues. Moreover, our design also provides a new model scheme for the development of two-photon fluorescent nanomaterial, which is expected to hold great potential for food processing and safety testing.

Diagnosis of bovine tuberculosis using a metal oxide-based electronic nose.

Electronic noses (e-noses) have been used for environmental monitoring, standardization of medicinal flavourings, food safety tests and diagnosis of infectious diseases based on the statistical analysis of volatile organic compounds (VOCs). Bovine tuberculosis (bTB) is officially diagnosed using the intradermal skin test (IST), which is time-consuming and labour-intensive. Therefore, a more convenient and rapid test with greater sensitivity would be advantageous as prescreening test. In this study, we used a metal oxide sensor (MOS) type e-nose to analyse VOCs in a bTB-infected (n = 11) and bTB-free (n = 10) sera, from cattle whose health status was confirmed using the IST, and pathological and bacteriological examinations. The differences in VOCs from bTB-infected and bTB-free sera detected by the e-nose were statistically analysed using principal components and discriminant factor analyses. bTB-infected and bTB-free sera could be discriminated by MOS type e-nose, and analysing time per sample was only 20 min. VOC analysis using a MOS e-nose was a rapid and automated prescreening method to diagnose bTB, and can be used to select bTB-suspect cattle for IST confirmation. Further studies are required to estimate test sensitivity and specificity. Significance and impact of the study: Bovine tuberculosis (bTB) in cattle is diagnosed using the intradermal skin test (IST); however, this method is very time-consuming and labour-intensive. We analysed volatile organic compounds that are obtained from serum using a metal oxide sensor type of electronic nose to discriminate between TB-infected and TB-free sera. This simple and automated technique will be useful to prescreen bTB-suspects and reduce the time and labour required to perform the IST.

Food safety assessment of an antifungal protein from Moringa oleifera seeds in an agricultural biotechnology perspective

Mo-CBP3 is an antifungal protein produced by Moringa oleifera which has been investigated as potential candidate for developing transgenic crops. Before the use of novel proteins, food safety tests must be conducted. This work represents an early food safety assessment of Mo-CBP3, using the two-tiered approach proposed by ILSI. The history of safe use, mode of action and results for amino acid sequence homology using the full-length and short contiguous amino acids sequences indicate low risk associated to this protein. Mo-CBP3 isoforms presented a reasonable number of alignments (>35% identity) with allergens in a window of 80 amino acids. This protein was resistant to pepsin degradation up to 2 h, but it was susceptible to digestion using pancreatin. Many positive attributes were presented for Mo-CBP3. However, this protein showed high sequence homology with allergens and resistance to pepsin digestion that indicates that further hypothesis-based testing on its potential allergenicity must be done. Additionally, animal toxicity evaluations (e.g. acute and repeated dose oral exposure assays) must be performed to meet the mandatory requirements of several regulatory agencies. Finally, the approach adopted here exemplified the importance of performing an early risk assessment of candidate proteins for use in plant transformation programs.

Iron (III)-Sodium-EDTA, as used for food fortification, aggravates intestinal inflammation and drives tumorigenesis in mouse models of colitis-associated cancer

Western lifestyle is associated with increasing prevalence of IBD, processed food and food fortification being a likely cause. Iron supplementation may increase gut inflammation and colitis-associated cancer (CAC). Novel Fe products which are chemically not Fe(II) salts and presumably better tolerated have become available. Here we compared various Fe compounds at 450 mg elemental Fe/kg chow [Na-EDTA-Fe(III) (as used for food fortification), Fe(III)maltol (Iron Therapeutics), plant Fe (Biogena), and Fe(II)SO4)], in their potential to promote colitis and CAC in animal models (AOM/DSS and IL-10-/-). In addition, in vitro experiments evaluated ROS production (DCFD assay), DNA damage (yH2AX) and oncogenic pathways in HT29, HCT116, Caco-2, RKO, and 1CT cells. Unexpectedly, Na-EDTA-Fe(III)-treated mice developed the severest form of colitis and only the 1st and last DSS cycles were administered. Similarly, in IL-10-/- mice, the Na-EDTA-Fe(III) group was euthanized 4 months before all other groups due to severe weight loss and disease activity (DAI). The mean DAI in the Na-EDTA-Fe(III) AOM/DSS mice was higher (1.82 vs. 0.94 (control), 0.76 (Fe(II)SO4), 0.67 (Fe(III)maltol), 0.54 (plant Fe), 0.80 (iron deficient); p < 0.001 by ANOVA). The total tumor area was higher with Na-EDTA-Fe(III) (5.96 mm2 vs. 1.38 mm2 (control), 1.88 mm2 (FeSO4), 1.97 mm2 (Fe(III)maltol), 0.98 mm2 (plant Fe), 1.51 mm2 (iron deficient); p = 0.001 by ANOVA) and there were more invasive tumors. ROS production was not elevated but there was an increase in yH2AX with Na-EDTA-Fe(III) and phosphorylation of Akt and Erk. Na-EDTA-Fe(III), as used for fortification in flour and cereals, exacerbates intestinal inflammation and drives tumorigenesis in mouse models of IBD. It is unclear whether this effect is caused by EDTA itself (which also is added to food as Na-EDTA or Na-EDTA-Ca(II)) or only in conjunction with Fe. Nevertheless, our data point to the hazard of food processing and fortification regarding IBD and to the necessity of food safety testing in animal models thereof.

Detection of shiga toxins by lateral flow assay.

Shiga toxin-producing Escherichia coli (STEC) produce shiga toxins (Stxs) that can cause human disease and death. The contamination of food products with STEC represents a food safety problem that necessitates rapid and effective detection strategies to mitigate risk. In this manuscript, we report the development of a colorimetric lateral flow assay (LFA) for the rapid detection of Stxs in <10 min using a pair of monoclonal antibodies that bind epitopes common to Stx1 and six Stx2 variants. This LFA provides a rapid and sensitive test for the detection of Stxs directly from STEC culture supernatants or at risk food samples with a 0.1 ng/mL limit of detection (LOD) for Stx2a. This Stx LFA is applicable for use in the rapid evaluation of Stx production from cultured E. coli strains or as a tool to augment current methods as part of food safety testing.

Electrical and electrochemical monitoring of nucleic Acid amplification.

Nucleic acid amplification is a gold standard technique for analyzing a tiny amount of nucleotides in molecular biology, clinical diagnostics, food safety, and environmental testing. Electrical and electrochemical monitoring of the amplification process draws attention over conventional optical methods because of the amenability toward point-of-care applications as there is a growing demand for nucleic acid sensing in situations outside the laboratory. A number of electrical and electrochemical techniques coupled with various amplification methods including isothermal amplification have been reported in the last 10 years. In this review, we highlight recent developments in the electrical and electrochemical monitoring of nucleic acid amplification.

Spectroscopic Techniques for Nondestructive Detection of Fungi and Mycotoxins in Agricultural Materials: A Review

Purpose: Fungal secondary metabolite (mycotoxin) contamination in foods can pose a serious threat to humans and animals. Spectroscopic techniques have proven to be potential alternative tools for early detection of mycotoxins. Thus, the aim of this review is to provide an overview of the current developments in nondestructive food safety testing techniques, particularly regarding fungal contamination testing in grains, focusing on the application of spectroscopic techniques to this problem. Methods: This review focuses on the use of spectroscopic techniques for the detection of fungi and mycotoxins in agricultural products as reported in the literature. It provides an overview of the characteristics of the main spectroscopic methods and reviews their applications in grain analysis. Results: It was found that spectroscopy has advantages over conventional methods used for fungal contamination detection, particularly when combined with chemometrics. These advantages include the rapidness and nondestructive nature of this approach. Conclusions: While spectroscopy offers many benefits for the detection of mycotoxins in agricultural products, a number of limitations exist, which must be overcome prior to widespread adoption of these techniques.

Method Validation for Qualitative and Quantitative Analysis of Pesticide Residues in Tomato with GC-MS/MS (TQD) for Food Safety Testing

Pesticide residues analysis in fruits and fresh vegetables is a challenge for food safety as the gap between pesticide sprays and harvests is very less in vegetables. A multi residue method was developed for qualitative and quantitative analysis of 64 pesticides (insecticides, fungicides and herbicides) using QuEChERS extraction method and GC-MS/MS (Triple Quadrupole) for analysis. Eight different concentrations of certified reference materials from 0.05 ppm to 0.30 ppm were injected in GC-MS/MS with MRM method, in six replications, and R2 ranged from 0.990–0.999 with RSD of 0.55 to 11.24. The sample preparation approach is through adoption of QuEChERS method, untreated control tomato samples were fortified with mixture of pesticides at 0.05, 0.25, 0.5 mg/kg, each is five replications, and the recovery of pesticides is in the range of 80–95%, and hence method can be used for qualitative and quantitative analysis of 64 pesticides in/on tomato for monitoring studies.