A Comparative Study on the Combustion Chemistry of Two Bio-hybrid Fuels: 1,3-Dioxane and 1,3-Dioxolane.

Bio-hybrid fuels are a promising solution to accomplish a carbon-neutral and low-emission future for the transportation sector. Two potential candidates are the heterocyclic acetals 1,3-dioxane (C4H8O2) and 1,3-dioxolane (C3H6O2), which can be produced from the combination of biobased feedstocks, carbon dioxide, and renewable electricity. In this work, comprehensive experimental and numerical investigations of 1,3-dioxane and 1,3-dioxolane were performed to support their application in internal combustion engines. Ignition delay times and laminar flame speeds were measured to reveal the combustion chemistry on the macroscale, while speciation measurements in a jet-stirred reactor and ethylene-based counterflow diffusion flames provided insights into combustion chemistry and pollutant formation on the microscale. Comparing the experimental and numerical data using either available or proposed kinetic models revealed that the combustion chemistry and pollutant formation differ substantially between 1,3-dioxane and 1,3-dioxolane, although their molecular structures are similar. For example, 1,3-dioxane showed higher reactivity in the low-temperature regime (500-800 K), while 1,3-dioxolane addition to ethylene increased polycyclic aromatic hydrocarbons and soot formation in high-temperature (>800 K) counterflow diffusion flames. Reaction pathway analyses were performed to examine and explain the differences between these two bio-hybrid fuels, which originate from the chemical bond dissociation energies in their molecular structures.

Uptake and distribution of difenoconazole in rice plants under different culture patterns.

The effects of spraying and root irrigation on the uptake and transport of the fungicide difenoconazole under hydroponic and soil cultivation were investigated. Rice was used as the crop for a short-term exposure experiment. A modified QuEChERS pre-treatment combined with ultra-high-performance liquid chromatography-tandem mass spectrometry was used to extract and detect difenoconazole from rice plants, water and soil. The recoveries of difenoconazole were in the range of 72.8-110.5%, with a relative standard deviation of 2.4-19.5% for all the samples when spiked with 0.01, 0.1 and 1 mg kg-1 of difenoconazole, respectively. The limit of quantitation (LOQ) of this method was 0.01 mg kg-1. The exposure results showed that difenoconazole could be absorbed by rice plants and transmitted to different parts of rice plants in all the treatments. In the hydroponic experiment, difenoconazole was mainly distributed in the roots of rice regardless of whether irrigation or spraying was used. For rice cultivated in soil, difenoconazole mainly accumulated in leaves after the root irrigation treatment, whereas after the spraying treatment, the rice roots were the main site of accumulation of difenoconazole. This experiment extends our knowledge of the influence of the cultivation system and application mode on the translocation of difenoconazole in rice plants.

Chromatographic analysis and residue degradation of phenamacril and difenoconazole on strawberries.

Strawberries are widely cultivated and highly consumed globally, but pests and diseases can severely affect yields. Phenamacril and difenoconazole are high-efficacy pesticides and the mixture of these two pesticides offers a satisfactory option for disease control. In this study, an optimised QuEChERS method combined with dispersive solid-phase extraction purification before injection for simultaneously determining the residues of phenamacril-difenoconazole mixture on strawberries was developed and validated using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Average recoveries of phenamacril and difenoconazole in the strawberry matrices ranged from 100% to 104% and 99% to 104%, with relative standard deviations of 2.6%-5.3% and 2.2%-5.5%, respectively. The degradation half-lives of phenamacril and difenoconazole were 3.5-6.6 days and 2.2-3.4 days on strawberries, respectively. Final residues of phenamacril and difenoconazole on strawberries at eight different cultivation regions were 0.033-0.66 mg kg-1 and <0.02-0.089 mg kg-1 after spraying at the maximum dosage recommended by the company of 300 mg a.i. kg-1 twice, respectively. Overall, this study is the first report of phenamacril and difenoconazole residue analysis in strawberries. Therefore, it could provide the reference data for safe management and proper use of phenamacril and difenoconazole in China.

New Insight into Justicidin B Pathway and Production in Linum austriacum.

Lignans are the main secondary metabolites synthetized by Linum species as plant defense compounds but they are also valuable for human health, in particular, for novel therapeutics. In this work, Linum austriacum in vitro cultures, cells (Cc), adventitious roots (ARc) and hairy roots (HRc) were developed for the production of justicidin B through elicitation with methyl jasmonate (MeJA) and coronatine (COR). The performances of the cultures were evaluated for their stability, total phenols content and antioxidant ability. NMR was used to identify justicidin B and isojusticidin B and HPLC to quantify the production, highlighting ARc and HRc as the highest productive tissues. MeJA and COR treatments induced the synthesis of justicidin B more than three times and the synthesis of other compounds. RNA-sequencing and a de novo assembly of L. austriacum ARc transcriptome was generated to identify the genes activated by MeJA. Furthermore, for the first time, the intracellular localization of justicidin B in ARc was investigated through microscopic analysis. Then, HRc was chosen for small-scale production in a bioreactor. Altogether, our results improve knowledge on justicidin B pathway and cellular localization in L. austriacum for future scale-up processes.

Abuse of nutmeg seeds: Detectable by means of liquid chromatography-mass spectrometry techniques?

Numerous case reports of intoxications with nutmeg seeds (Myristica fragrans, Houtt.) can be found in literature often following their abuse, as psychotropic effects were described after ingestions of large doses. The successful detection of the main ingredients of the nutmeg seeds essential oil elemicin, myristicin, and safrole, as well as their metabolites in human urine by gas chromatography coupled to mass spectrometry (GC-MS) was already described. The aim of this study was to investigate the detectability of the main ingredients of nutmeg seeds and their metabolites in human blood and urine samples using liquid chromatography coupled to linear ion trap mass spectrometry (LC-LIT-MSn ) and liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS/MS) after nutmeg seed abuse. Sample material of three individuals was retrospectively investigated after a systematic screening approach indicated an intoxication with nutmeg seeds as a likely cause of symptoms. Metabolic patterns in plasma and urine using GC-MS were comparable with those described in earlier publications. Investigations using hyphenated liquid chromatography techniques lead to the detection of myristicin and safrole, as well as further metabolites not described using GC-MS and revealed sulfation as an additional Phase II metabolic pathway. These results might help to detect or confirm future intoxications with nutmeg seeds by using LC-MS techniques.

Statistical evaluation of analytical curves for quantification of pesticides in bananas.

The aim of this paper is to statistically validate the analytical curves of a chromatography method to identify and quantify azoxystrobin, difenoconazole and propiconazole residues in banana pulp, using QuEChERS and GC-SQ/MS. A matrix-matched calibration was used and analytical curves were estimated by weighted least squares regression (WLS), confirming heteroscedasticity for all compounds. Statistical tests were performed to confirm the quality adjustment of the proposed linear model. The correlation coefficient for azoxystrobin, difenoconazole and propiconazole were, respectively, 0.9985, 0.9966 and 0.9997 (concentration range: 0.05 and 2.0 mg kg-1). The limits of detection and quantification were, respectively, between 0.007 and 0.066 mg kg-1, and between 0.022 and 0.199 mg kg-1, below the maximum limits stipulated by Brazilian, American, and European legislation. Only difenoconazole had an insignificant matrix effect (6.8%). Thus, the weighted least squares method is shown to be a safe linear regression model, providing greater reliability of the results.

Exhaled breath analysis using on-line preconcentration mass spectrometry for gastric cancer diagnosis.

Breath volatile biomarkers are capable of distinguishing patients with various cancers. However, high throughput analytical technology is a prerequisite to a large-cohort study intended to discover reliable breath biomarkers for cancer diagnosis. Single-photon ionization (SPI) is a universal ionization technology, and SPI-mass spectrometry (SPI-MS) shows a remarkable advantage in the comprehensive detection of volatile organic compounds (VOCs), in particular, nonpolar compounds. In this study, we have introduced SPI-MS coupled with on-line thermal desorption (TD-SPI-MS) to demonstrate nontarget analysis of breath VOCs for gastric cancer patients. The breath fingerprints of the gastric cancer patients were significantly distinct from that of the control group. Acetone, isoprene, 1,3-dioxolan-2-one, phenol, meta-xylene, 1,2,3-trimethylbenzene, and phenyl acetate showed higher relative peak intensities in the breath profiles of gastric cancer patients. A diagnostic prediction model was further developed by using a training set (121 samples) and validated with a test set (53 samples). The predication accuracy of the developed model was 96.2%, and the area under the curve (AUC) of the receiver operator characteristic curve (ROC) was 0.997, indicating a satisfactory prediction ability of the developed model. Thus, by taking gastric cancer as an example, we have shown that TD-SPI-MS will be a promising tool for high throughput analysis of breath samples to discover characteristic VOCs in patients with various cancers.

Competitive Adsorption and Mobility of Propiconazole and Difenoconazole on Five Different Soils.

Propiconazole (PPC) and difenoconazole (DFC) are often combined for field applications. The resulted co-exsistence of PPC and DFC may have an effect on the fate of their individuals in soil. In this study, adsorption, desorption and leaching of PPC and DFC alone and their combinations were investigated in five different soils. Adsorption of PPC and DFC was significantly different on each soil with the Freundlich adsorption coefficients of 2.86-28.69 and 14.86-98.93 negatively correlated with soil pH, respectively. In addition, adsorption of PPC and DFC was declined by 27.12-37.59% and 17.28-25.35% with the presence of coexisting DFC and PPC, respectively. Mobility of PPC and DFC in tested soils was enlarged in coexisting system. The results indicate that adsorption, desorption and mobility of PPC and DFC were mainly affected by soil pH, and these behaviors of individual PPC and DFC were obviously altered by their co-existence.

Salting-out assisted liquid-liquid extraction method optimized by design of experiments for the simultaneous high-performance liquid chromatography analysis of perampanel and stiripentol in mouse matrices.

We report a high-performance liquid chromatography method development able to simultaneously determine perampanel and stiripentol, two third-generation antiepileptics whose therapeutic spectrum can potentially be extended, in several mouse matrices. A salting-out assisted liquid-liquid extraction optimized by a design of experiments approach was adopted for sample preparations. Isopropanol and magnesium sulfate were the extraction solvent and salting-out agent, respectively. Both drugs and internal standard (terbinafine) were separated using a LiChroCART® Purospher Star column (C18 , 55 × 4 mm; 3 µm) isocratically pumped with mobile phase [1% triethylamine in water (pH 2.5) and acetonitrile (53:47, v/v)] at 1 mL/min. Stiripentol and terbinafine were detected by fluorescence at 254/372 nm and perampanel at 275/430 nm. Good linearity was demonstrated for perampanel at 1-500 ng/mL range in brain, 2-2000 ng/mL in liver and 1-2000 ng/mL in plasma and kidney (r2  ≥ 0.9922), and for stiripentol between 10 and 2000 ng/mL in brain and 10 and 20 000 ng/mL in the remaining matrices (r2  ≥ 0.9917). Precision (CV ≤ 15%) and accuracy (bias ±15%) were also verified, with obtained recovery values consistent with those predicted by the experimental design. This method was applied in preliminary pharmacokinetic studies to quantify perampanel or stiripentol after oral administration to mice, showing to be a promising bioanalytical tool to support future nonclinical in vivo pharmacokinetic studies.

A new strategy based on gas chromatography-high resolution mass spectrometry (GC-HRMS-Q-Orbitrap) for the determination of alkenylbenzenes in pepper and its varieties.

Alkenylbenzenes are natural toxins with genotoxic and carcinogenic effects in rodents, which are highly present in condiments frequently consumed. The aim of this study was the development of the first multi-analyte method for the determination of eight alkenylbenzenes (eugenol, methyl eugenol, acetyl eugenol, trans-isoeugenol, safrole, estragole, myristicin and trans-anethole) in different pepper varieties by gas chromatography coupled to high-resolution mass spectrometry (GC-HRMS-Q-Orbitrap) in combination with a simple ultrasound-assisted extraction method (UAE). The method was successfully validated, and it was applied for studying the presence of these analytes in peppers as well as to elucidate the effects of the berries' maturity and the geographical origin on alkenylbenzene contents. The analysis of the pepper samples showed that eugenol (10.5-120 mg/kg), trans-anethole (10.7-42.7 mg/kg) and estragole (2.2-45.7 mg/kg) tended to be the most detected alkenylbenzenes at high levels, whereas trans-isoeugenol (0.69-3.6 mg/kg) and safrole (0.20-3.0 mg/kg) were minor components. Estragole (PubChem CID: 8815); trans-anethole (PubChem CID: 637563); Myristicin (PubChem CID: 4276); Safrole (PubChem CID: 5144); Eugenol (PubChem CID: 3314); Methyl eugenol (PubChem CID: 7127); Acetyl eugenol (PubChem CID: 7136); trans-Isoeugenol (PubChem CID: 853433); Caffeine (PubChem CID: 2519); Dicyclohexylmethanol (PubChem CID: 78197).

Detection of Nutmeg Abuse by Gas Chromatography-Mass Spectrometric Screening of Urine.

High doses of nutmeg (seeds from Myristica fragrans Houtt.) can be abused as a psychoactive drug due to phenylpropene ingredients. During controlled abstinence, e.g., in forensic psychiatric clinics, nutmeg abuse has to be distinguished from an ingestion of other spices having phenylpropene ingredients (e.g., black pepper or garden lovage) or unintentional low-dose nutmeg intake. The aim of this study was to develop an evaluation model for the estimation of time point and amount of nutmeg abuse and differentiation from ingestion of other spices or low doses of nutmeg based on the gas chromatographic-mass spectrometric (GC-MS) analysis of urine samples. A total of 3 volunteers ingested 1.5 g of freshly ground nutmeg. No symptoms were reported. Urine samples were collected for up to 3 days. In addition, 18 blank samples from volunteers with regular diet and 2 authentic samples from forensic psychiatry patients with supposed nutmeg abuse were analyzed. All samples were analyzed by GC-MS in full scan mode. Metabolites of the nutmeg ingredients safrole, myristicin and elemicin were identified via a library search. For semi-quantitative estimations, the area ratios of the analytes to the internal standard (MDMA-d5) were normalized to the creatinine concentration. Up to 8 different metabolites were detected for at least 18 hours after intake of 1.5 g of nutmeg. In the two authentic samples, the normalized area ratios of those metabolites were 0.5-14 times the maximum reached in the intake study. Two additional metabolites could be detected in authentic samples. Probably due to ingestion of other spices, 5 of the 8 metabolites after intake of 1.5 g of nutmeg were detected in blank urine samples as well. The intake of high doses of nutmeg can be differentiated from the ingestion of other spices or low doses of nutmeg via standard GC-MS analysis of urine and application of the proposed evaluation model.

Ocotea pulchellaas an alternative against schistosomiasis: chemical analysis, development of nanoemulsion and biological control activity / Ocotea pulchellacomo una alternativa ante esquistosomiasis: análisis químico, desarrollo de una nanoemulsión y actividad en control biológico

The aim of this work was to evaluate the potential of the essential oil (EO) from Ocotea pulchella leaves as an alternative in the control of schistosomiasis. It was tested O. pulchella EO nanoformulation to assess its activity against adult Biomphalaria glabrata, their spawning and Schistossoma mansoni cercariae. Additionally, the EO chemical composition was investigated by gas-chromatography. Nanoemulsion were elaborated by the low energy method. The adult mollusks, their spawning and cercariae were placed in contact with nanoemulsion to calculate lethal concentrations. Myristicin, bicyclogermacrene and α-Pinene were the main substances in the EO. Nanoemulsion caused mortality of adult B. glabrata, its egg embryos and S. mansoni. These results suggest the use of this nanoemulsion as an alternative in the control of the schistosomiasis cycle.

El objetivo de este trabajo fue evaluar el potencial de los aceites esenciales (AE) de las hojas de Ocotea pulchellacomo una alternativa en el control de esquistosomiasis. Se probó una nanoformulación de AE de O. pulchellapara evaluar su actividad ante adultos de Biomphalaria glabrata, sus huevos y cercarías de Schistossoma mansoni. La nanoemulsión fue elaborada por el método de baja energía. Los moluscos adultos, sus huevos y cercarías se colocaron en contacto con la nanoemulsión para calcular concentraciones letales. Los compuestos mayoritarios en el AE fueron miristicina, biciclogermacreno y α-pineno. La nanoemulsión causó mortalidad en adultos de B. glabrata, sus huevos y a S. mansoni. Los resultados sugieren el uso de esta nanoemulsión como una alternativa en el control del ciclo de esquistosomiasis.

Use of ozone and detergent for removal of pesticides and improving storage quality of tomato.

The efficiencies of two conventional domestic procedures (immersion in pure water and detergent solution at 0.25 and 1%) and two treatments using ozone (immersion in water with bubbling O3 and immersion in ozonated water, both at 1 and 3 mg L-1) were evaluated for the removal of residues of the fungicides azoxystrobin, chlorothalonil and difenoconazole in tomatoes. The fungicides were sprayed on the fruits at the recommended concentration for the crop. The residues in the tomatoes and in the washing solutions were determined by extraction with low-temperature partition techniques and analysis by gas chromatography. More concentrated solutions were more effective in removing pesticide residues. The water bubbled with ozone at 3 mg L-1 was the most efficient treatment for the removal of fungicides, reaching a reduction of 70-90% of the residues. However, the treatments with the lowest concentration of ozone had lower loss of fruit mass during storage.

Dissipation rates and residue levels of diflubenzuron and difenoconazole on peaches and dietary risk assessment.

The dissipation kinetics, residue levels, and potential risks of diflubenzuron and difenoconazole on peaches were investigated under open field conditions. Two years of field trials were carried out in Shanghai, China, and the half-lives of diflubenzuron and difenoconazole on peaches ranged from 4.4 to 25d. Their terminal residue concentrations on peaches were 0.022-5.7 mg/kg after three of the tested sampling intervals. Based on the maximum residue levels (MRLs) of difenoconazole on peaches, a preharvest interval (PHI) of 14 d was proposed. A PHI of 10 d was proposed for diflubenzuron after a dietary safety assessment. During the safety assessment, the hazard quotient (HQ) and risk quotient (RQ) on peaches were determined. The results showed that the HQs (3.6-8.3%) and RQs(51-55%) of diflubenzuron were acceptable, proving that diflubenzuron poses no potential health risks. For difenoconazole, the HQs (0.027-0.071%) were satisfactory, but the RQs (115-116%) exceeded 100%, which indicated potential risk.

HS-SPME analysis of the volatiles profile of water celery (Apium nodiflorum), a wild vegetable with increasing culinary interest.

Water celery (Apium nodiflorum) is a wild plant traditionally harvested in some Mediterranean areas for being consumed raw. Despite its appreciated organoleptic properties, the aromatic profile of the fresh vegetable remains to be studied. In the present study, volatile compounds from five wild populations were extracted by the headspace-solid phase microextraction technique, analysed by gas cromatography-mass spectrometry, and compared to related crops. The wild species had a high number of aromatic compounds. It was rich in monoterpenes (49.2%), sesquiterpenes (39.4%) and phenylpropanoids (9.6%), with quantitative differences among populations, in absolute terms and relative abundance. On average, germacrene D was the main compound (16.6%), followed by allo-ocimene (11.9%) and limonene (11.1%). Only in one population, the levels of limonene were greater than those of germacrene D. Among phenylpropanoids, dillapiol displayed the highest levels, and co-occurred with myristicin in all populations except one. These differences may have a genetic component, which would indicate the possibility of establishing selection programmes for the development of water celery as a crop adapted to different market preferences. On the other hand, comparison with related crops revealed some similarities among individual volatiles present in the different crops, which would be responsible of the common aroma notes. However, water celery displayed a unique profile, which was in addition quantitatively richer than others. Thus, this differentiation may promote the use of water celery as a new crop.

Essential oil composition of aerial parts from Algerian Anacyclus monanthos subsp. cyrtolepidioides (Pomel) Humphries.

The chemical composition of the essential oil from the aerial parts of Anacyclus monanthos subsp. cyrtolepidioides (Pomel) Humphries (Asteraceae) growing in a semi-arid region of Algeria was investigated for the first time. The essential oil was obtained by hydrodistillation and fully characterized by gas chromatography-mass spectrometry (GC-MS). A total of 97 compounds were identified. The essential oil was found to be rich in trans-chrysanthenyl acetate (9.8 ± 2.0%), (E)-ß-farnesene (7.4 ± 1.5%), germacrene D (6.9 ± 1.3%) and myristicin (4.8 ± 0.8%).

Development of a method for quantitative determination of the cytotoxic agent piplartine (piperlongumine) in multiple skin layers.

This study reports the development of a simple and reproducible method, with high rates of recovery, to extract the cytotoxic agent piplartine from skin layers, and a sensitive and rapid UV-HPLC method for its quantification. Considering the potential of piplartine for topical treatment of skin cancer, this method may find application for formulation development and pharmacokinetics studies to assess cutaneous bioavailability. Porcine skin was employed as a model for human tissue. Piplartine was extracted from the stratum corneum (SC) and remaining viable skin layers (VS) using methanol, vortex homogenization and bath sonication, and subsequently assayed by HPLC using a C18 column, and 1:1 (v/v) acetonitrile-water (adjusted to pH 4.0 with acetic acid 0.1%) as mobile phase. The quantification limit of piplartine was 0.2 µg/mL (0.6 µm), and the assay was linear up to 5 µg/mL (15.8 µm), with within-day and between-days assay coefficients of variation and relative errors <15%. Piplartine recovery from SC and VS varied from 86 to 96%. The method was suitable to assay samples from skin penetration studies, enabling detection of differences in cutaneous delivery in different skin compartments resulting from treatment with various formulations and time periods.

Determination of azole fungal residues in soils and detection of Aspergillus fumigatus-resistant strains in market gardens of Eastern France.

Medical azole antifungals are major compounds used to prevent and to treat invasive aspergillosis (IA). Azole fungicides, called DMI (14-alpha demethylase inhibitors), are also widely used for crop protection and have been reported to be linked to azole-resistant A. fumigatus (aR-Af) development in the environment. The aim of this study was to determine whether or not market gardens that spray DMIs in Eastern France are also affected by the presence of aR-Af. Forty aR-Afs were detected in soils in only two of the four market gardens using DMIs, with 23% (7/30) and 10% (3/30) of soils containing aR-Af. A total of 87.5% of these isolates had the TR34/L98H mutation and 22.5% the TR46/Y121F/T289A mutation on the cyp51A gene. Analyses of residual azole concentrations in soils showed the presence of difenoconazole for up to 2 years after spraying, but only in soils of market gardens where aR-Af was detected. It is very important to identify professional activities that could lead to aR-Af development and to develop preventive measures for at-risk patients living near professional activities using DMIs. We have to better understand why, in some cases, the use of DMI is not linked to aR-Af. Measures should be taken to avoid the use of DMI conferring cross-resistance to preserve the efficiency of human therapeutics.

Matrix Effect Evaluation and Method Validation of Azoxystrobin and Difenoconazole Residues in Red Flesh Dragon Fruit (Hylocereus polyrhizus) Matrices Using QuEChERS Sample Preparation Methods Followed by LC-MS/MS Determination.

Production of red flesh dragon fruit (Hylocereus polyrhizus) was hampered by Colletotrichum sp. Pre-harvest application of azoxystrobin and difenoconazole mixture is recommended, therefore, a selective and sensitive multi residues analytical method is required in monitoring and evaluating the commodity's safety. LC-MS/MS is a well-established analytical technique for qualitative and quantitative determination in complex matrices. However, this method is hurdled by co-eluted coextractives interferences. This work evaluated the pH effect of acetate buffered and citrate buffered QuEChERS sample preparation in their effectiveness of matrix effect reduction. Citrate buffered QuEChERS proved to produce clean final extract with relative matrix effect 0.4%-0.7%. Method validation of the selected sample preparation followed by LC-MS/MS for whole dragon fruit, flesh and peel matrices fortified at 0.005, 0.01, 0.1 and 1 g/g showed recoveries 75%-119%, intermediate repeatability 2%-14%. The expanded uncertainties were 7%-48%. Based on the international acceptance criteria, this method is valid.

[Determination of myclobutanil and difenoconazole residues in pollen and honey of litchi by high performance liquid chromatography-tandem mass spectrometry].

An effective method was developed for the determination of two major fungicides including myclobutanil and difenoconazole residues in pollen and honey of litchi by modified QuEChERS-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The pollen and honey samples were all extracted by acetonitrile, the pollen samples were cleaned-up by 0.9 g anhydrous magnesium sulfate (MgSO4), 0.15 g primary secondary amine (PSA) and 0.15 g C18; the honey samples were cleaned-up by 0.9 g MgSO4 and 0.15 g PSA. The 0.1% (v/v) formic acid aqueous solution-acetonitrile (25:75, v/v) were used as the mobile phases. The extracts were separated on a Poroshell-120 EC-C18 chromatographic column, the positive electrospray ion (ESI+) source and selected ion monitoring (SIM) mode were used. The analytes were quantified by the matrix matching standard solutions. The matrix matched standard solutions of myclobutanil and difenoconazole showed good linearities in the range of 1-100 µg/L, and the correlation coefficients (r2) were all above 0.9990. The limits of detection (LODs) of myclobutanil and difenoconazole were 0.25 µg/kg and 0.50 µg/kg, respectively. The limits of quantification (LOQs) of myclobutanil and difenoconazole were 0.83 µg/kg and 1.7 µg/kg, respectively. The average recoveries of myclobutanil and difenoconazole in pollen and honey samples were 87.0%-95.2% and 90.1%-96.4% with the relative standard deviations of 1.2%-3.6% and 0.7%-4.1%, respectively. The method is quick, easy and sensitive, and it is suitable for the rapid determination and trace analysis of myclobutanil and difenoconazole in pollens and honeys of litchi. The method can provide data support for the exposure risk assessment of bees and other pollination insects.