Pesticide residue analysis of fruit juices by LC-MS/MS direct injection. One year pilot survey

For this work, thirteen types of fruit juices (orange, pineapple, peach, apple, multifruit, mango, strawberry, tomato, pear, mandarin, grape, banana and grapefruit) were selected to develop an analytical method for the analysis of 53 pesticides by direct injection in LC-MS/MS. The preparation of the samples was very simple: an aliquot of the juice was centrifuged and it was ten-times diluted prior to analysis, which allowed reducing considerably the time and cost of the analyses. Besides, dilution of the samples permits reducing the amount of matrix going into the system, and thus, decreasing the matrix effects, so common in this type of commodities, opening the possibility to perform quantification with solvent based standards. Validation of the method was carried out in accordance with EU guidelines. Calibration curves covering three orders of magnitude were performed, and they were linear over the concentration range studied for all the matrices (from 0.1 to 100 μg L⁻¹). Practical limits of quantification were in the low μg L⁻¹ range, far below the maximum residue levels (MRLs) of the EU regulations, which do not set specific MRLs for juices, and in this cases of processed food, MRLs of the raw product are applied. Repeatability of the instrumental method was studied in all matrices, obtaining good intra- and inter-day relative standard deviations (RSDs). The proposed method was applied to 106 real fruit juice samples purchased in different local markets during a one-year survey in order to validate the suitability for routine analysis. 43% of the analysed samples gave positive results (higher than the practical limits of quantification).     

Atmospheric pressure glow discharge desorption mass spectrometry for rapid screening of pesticides in food

Flowing afterglow atmospheric pressure glow discharge tandem mass spectrometry (APGD-MS/MS) is used for the analysis of trace amounts of pesticides in fruit juices and on fruit peel. The APGD source was rebuilt after Andrade et al. (Andrade et al., Anal. Chem. 2008; 80: 2646-2653; 2654-2663) and mounted onto a hybrid quadrupole time-of-flight mass spectrometer. Apple, cranberry, grape and orange juices as well as fruit peel and salad leafs were spiked with aqueous solutions containing trace amounts of the pesticides alachlor, atrazine, carbendazim, carbofuran, dinoseb, isoproturon, metolachlor, metolcarb, propoxur and simazine. Best limits of determination (LODs) of pesticides in the fruit juices were achieved for metolcarb (1 microg/L in apple juice), carbofuran and dinoseb (2 microg/L in apple juice); for the analysis of apple skin best LODs were 10 pg/cm(2) of atrazine, metolcarb and propoxur which corresponds to an estimated concentration of 0.01 microg/kg apple, taking into account the surface area and the weight of the apple. The measured LODs were within or below the allowed maximum residue levels (MRLs) decreed by the European Union (1-500 microg/kg for pesticides in fruit juice and 0.01-5 microg/kg for apple skin). No sample pretreatment (extraction, pre-concentration, chromatographic separation) was necessary to analyze these pesticides by direct desorption/ionization using APGD-MS and to identify them using MS/MS. This makes APGD-MS a powerful high-throughput tool for the investigation of very low amounts of pesticides in fruit juices and on fruit peel/vegetable skin. Copyright (c) 2008 John Wiley & Sons, Ltd.     

Quantitative determination of limonin in citrus juices by HPLC using computerized solvent optimization

The commercially available computer program, Drylab, for optimization of separations by high-performance liquid chromatography (HPLC) using binary solvent mixtures is used to improve an HPLC method for separation of the bitter principle, limonin, in grapefruit and navel orange juices. Best conditions for separation of limonin in a reasonable time are 30 to 32% acetonitrile in water at 0.9 mL/min using a 5-micron C18 column 10 cm long. These conditions are used to analyze grapefruit and navel orange juice samples, and these HPLC results are compared with values determined by enzyme immunoassay or thin-layer chromatography (TLC) on the same samples.     

基于代谢组学技术的快速蒸发电离质谱与智能手术刀联用实时鉴别果汁掺伪方法研究

近年来,果汁掺伪问题日益突出。作为一种新型的实时鉴别方法,智能手术刀(IKnife)与快速蒸发电离质谱(REIMS)联用技术无需样品前处理即可解决这一问题。该文采用代谢组学指纹识别技术建立橙汁、苹果汁和葡萄汁的实时鉴别方法,并对其掺假水平进行预测。主成分分析-线性判别分析联用法建立的判别模型能够区分不同类型的果汁,排除20%交叉验证法的正确率为97.28%,未知样品实时鉴别正确率高达100%。筛选出不同果汁中的标志物,并对部分化合物进行鉴定。结果表明,此方法能够成功区分掺有10%-50%苹果汁和葡萄汁的橙汁。通过对比分析REIMS单极质谱和串联质谱(REIMS/MS)的检测结果,两种方法建立的偏最小二乘分析模型均能准确预测掺假水平(所有模型的R 2和Q 2均大于0.82)。与传统的REIMS方法相比,REIMS/MS的预测值更加精确。     

Contactless conductivity detection of sodium monofluoroacetate in fruit juices on a CE microchip

Rapid and quantitative determination of sodium monofluoroacetate in diluted fruit juices (dilution 1:9 v/v in deionized water) and tap water was performed by microchip CE, using contactless conductivity detection. A separation buffer consisting of 20 mM citric acid and histidine at pH 3.5 enabled the detection of the monofluoroacetate (MFA) anion in diluted apple juice, cranberry juice, and orange juice without lengthy sample pretreatments. The analyte was very well separated from interfering anionic species present in juices and tap water. LODs in diluted juices and tap water were determined to be 125, 167, 138, and 173 microg/L for tap water, apple juice, cranberry juice, and orange juice, respectively, based upon an S/N of 3:1. Taking into account the dilution factor, the LODs for juice samples range from 1 to 2 mg/L, which is adequate for monitoring the toxicity of MFA in these juice beverages and tap water. The calibration curves for MFA in diluted fruit juices were linear over the range of 500 microg/L to 80 mg/L. The total analysis time for detecting the MFA anion in fruit juices was less than 5 min, which represents a considerable reduction in analysis time compared to other analytical methods currently used in food analysis.     

超高效液相色谱-串联质谱法快速测定浓缩果汁中噻菌灵和多菌灵的残留量

建立了同时测定浓缩果汁中噻菌灵和多菌灵残留的超高效液相色谱-串联质谱快速检测法。样品用乙酸乙酯提取,以ACQUITY UPLC BEH C18色谱柱(50 mm×2.1 mm, 1.7 μm)进行超高效液相色谱分离,以电喷雾电离串联质谱正离子多反应监测(MRM)模式进行测定,以基质匹配标准溶液外标法定量。结果表明:在试验条件下,噻菌灵和多菌灵在0.5~10 μg/kg范围内线性关系良好,相关系数大于0.99,不同基质中的检出限(S/N=3)范围为0.12~0.23 μg/kg。在0.5、1.0和5.0 μg/kg 3个水平下噻菌灵和多菌灵的加标回收率为76.98%~108.7%,相对标准偏差(RSD)为2.95%~9.99%。同时,本研究对浓缩果汁中噻菌灵和多菌灵残留检测的基质效应进行了考察。本方法具有操作简便、快速、准确的特点,可用于浓缩果汁中噻菌灵和多菌灵残留量的日常检测。     

Polygalacturonase and ethanol production in Kluyveromyces marxianus

The coproduction of ethanol and polygalacturonase (PG) in a pilot-scale batch fermentor using yeast extract—glucose (YD)—and sugar beet molasses (SBM)-based media was implemented utilizing a new high-PG-producing strain of Kluveromyces marxianus. A certain growth inhibition was observed in SBM medium, causing ethanol and PG production to be lower. Ethanol productivity and accumulation values of 1.94 g/(L·h) and 40 g/L, respectively, were attained in YD, whereas the best fermentation efficiency (95.1%) was achieved with SBM medium. Maximal PG synthesis occurred at the end of cell growth, with values of 1.08 and 0.46 U/(mg·h) for the YD and SBM media, respectively. When the cultures reached stationary phase, PG production stopped. The highest accumulation level (17 U/mL) occurred in YD medium, in agreement with previous laboratory-scale studies carried out for this strain. The potential applications of the crude enzyme preparations were evaluated with different fruit juices and vegetable slices. The enzyme was able to increase the filtration rate of orange, pear, and apple juices by twofold. Additionally, complete clarification of apple juice was readily accomplished, whereas cucumber, carrot, and banana tissues were macerated to a lesser extent.     

Mixed-mode solid-phase extraction and cleanup procedures for the liquid chromatographic determination of thiabendazole and carbendazim in fruit juices

Solid-phase extraction (SPE) procedures were developed for rapid cleanup and determination of thiabendazole and carbendazim in orange, apple, and grape juices. Samples were prepared by using an SPE cartridge containing a mixed-mode sorbent with both reversed-phase and strong cation-exchange chemistries. Analysis was by liquid chromatography with photodiode-array UV detection. Orange juice was analyzed by mixed-mode cation-exchange extraction with reversed-phase cleanup; the other juices were analyzed by reversed-phase extraction with cation-exchange cleanup. Recoveries >80% for carbendazim and >90% for thiabendazole. Quantitation limits were 20 microg/L for both analytes.     

Application of a mixed-mode solid-phase extraction and cleanup procedure for LC/MS determination of thiabendazole and carbendazim in apple juice

Recently, a mixed-mode solid-phase extraction (SPE) procedure was developed for rapid extraction and cleanup for determination of the fungicides thiabendazole and carbendazim in various fruit juices. This paper reports the application of that sample preparation procedure to the liquid chromatographic/mass spectrometric determination of these fungicides in apple juice with detection by positive electrospray ionization mass spectrometry (ESI/MS). Response was linear for sample concentrations from 2 to 500 microg/L (ppb). Recoveries averaged 74% (9% RSD) for carbendazim and 93% (9% RSD) for thiabendazole. After SPE cleanup, no matrix supression was observed for the ESI+ response for either compound studied. The method was applied to the analysis of incurred residues in 4 store-bought apple juices; carbendazim levels ranged from 10 to 70 microg/L and thiabendazole levels ranged from less than 2 to 130 microg/L.     

Liquid chromatographic method for determination of patulin in clear and cloudy apple juices and apple puree: collaborative study

A collaborative trial was conducted to validate the effectiveness of a liquid chromatographic (LC) procedure for determination of patulin in both clear and cloudy apple juices and apple puree. The test portion of clear apple juice was directly extracted with ethyl acetate; cloudy apple juice and apple puree were treated with pectinase enzyme before extraction. After back-extraction into sodium carbonate to remove interfering acidic compounds, the extract was dried and concentrated, and patulin was determined by LC with UV detection. Clear and cloudy apple juices, apple puree test samples naturally contaminated with patulin, and blank test samples for spiking with patulin were sent to 14 collaborators in 12 different European countries. Test portions of each of the 3 test sample types were spiked with patulin at 75 ng/g. Recoveries of patulin ranged from 80 to 92%. Based on the results for spiked test samples (blind pairs) and naturally contaminated test samples (blind pairs at 3 levels), the relative standard deviations for repeatability (RSDr) and reproducibility (RSDR) ranged from 8 to 35% and 11 to 36%, respectively. Although HORRAT values of <1.4 were obtained for all 3 matrixes at patulin levels ranging from 26 to 121 ng/g, better performance values (RSDr values 6-10% and RSDR values 11-25%) were obtained for clear and cloudy apple juice spiked above 50 ng/g, which is either the statutory limit or the advisory level for patulin contamination in apple juices in many countries.     

Production of High-Quality Red Fruit Juices by Athermal Membrane Processes

Membrane-based processes are increasingly used to clarify and concentrate thermo-sensitive fruit juices and plant extracts as alternatives to conventional processes. This work aimed to evaluate the quality of red fruit juices clarified and concentrated by an integrated membrane process with special regard to the preservation of valuable compounds. A red fruit juice obtained from a blend of pomegranate, cactus pear, and red orange juices of Sicilian origin was clarified by microfiltration (MF) and then pre-concentrated up to 33 °Brix by nanofiltration (NF). The pre-concentrated juice was finally concentrated by osmotic distillation (OD) up to 50 and 60 °Brix. Samples of clarified, pre-concentrated, and concentrated juice were analyzed for their physico-chemical composition and in terms of the antioxidant activity and inhibitory activity against α-amylase and lipase. The results clearly confirmed the assumption of a mild fruit juice processing method, allowing us to preserve the original nutritional and functional properties of the fresh juice. In particular, the OD retentate at 60 °Brix resulted the most active sample against pancreatic lipase and α-amylase inhibitory activity with IC₅₀ values of 44.36 and 214.65 μg/mL, respectively.     

The Intehaction of Aqueous Solutions of Chlorine with Malic Acid,Tartaric Acid,and Various Fruit Juices: A Source of Mutagens

Abstract

The interactions of aqueous solutions of chlorine with some fruit acids (citric acid, DL-malic acid, arid L-tartaric acid) at different pH values were studied. Diethyl ether extraction followed by GC/MS analysis indicated that a number of mutagens (certain chlorinated propanones and chloral hydrate) are present as major products in some of these samples. A number of fruit juices (orange, grape, apple, pineapple, and grapefruit) were also treated with aqueous solutions of chlorine at their pH values. The products were analyzed by GC/MS. The same mutagens that were formed by the pure acids (citric acid and DL-malic acid) were identified as major products in ether extracts of these samples. Another mutagen, dichloroacetonitrile, was also identified as a minor product in some of these juice samples. All of the major products observed in the chlorination of all five fruit juices are potentially derived from reactions of aqueous solutions of chlorine with citric or malic acid and with trace amounts of acetaldehyde and acetone in the juices. The minor product, dichloroacetonitrile, is likely derived from the chlorination of certain amino acids in the fruit juices.     

Determination of trace cobalt in fruit samples by resonance ionization mass spectrometry

1-color 2-photon resonance ionization mass spectrometry (RIMS) with a new electro-thermal atomizer equipped with six filaments has been applied for quantitative analysis of trace cobalt in fruit samples. The efficient transition lines for sensitive detection were investigated in the wavelength range of 283-302 nm and a transition at 298.71 nm was chosen as an excited state for the determination of Co. A proper calibration curve for cobalt up to 1 ppb has been constructed with satisfactory results. The minimum detectible amount of cobalt in this RIMS system was identified as less than 5 pg. The content of Co in three different fruits, pear, apple and Korean mandarin orange, have been determined by adopting standard addition to the fruit sample juice. The content of the Co in 5 μl of apple, pear and Korean mandarin juice was identified as 150, 45 and 100 pg, respectively.     

High-performance liquid chromatographic analysis of free amino acids in fruit juices using derivatization with 9-fluorenylmethyl-chloroformate

A simple, rapid, and reliable reversed-phase high-performance liquid chromatographic method for the analysis of 16 amino acids of main interest in commercial fruit juices (pear, orange, grapefruit, pineapple, peach, and apricot) is described. No sample cleanup is required. The pH of the fruit juices is adjusted to alkaline value (8.5) using 200 mM borate buffer, then amino acid is converted to stable derivatives using 9-fluorenylmethyl-chloroformate. The excess of derivatization reagent is removed by a hydrophobic amine, 1-amino-adamantane hydrochloride. The derivatization procedure is simple, fast, and described in detail. Amino acids are detected at 263 nm and eluted within 35 min. The calibration, precision (< or = 6.1%), and recovery (102% +/- 4%) of the method are reported. The conditions of separation are optimized; however, serine partially overlapped with aspartic acid. The amino acid profile of fruit juices is consistent with data from the literature.     

Application of graphene for the SPE clean‐up of organophosphorus pesticides residues from apple juices

In this paper, an effective graphene‐based SPE clean‐up procedure coupled with GC–MS was developed for the determination of organophosphorus pesticide residues in apple juices. The apple juice samples were diluted with water and could be loaded onto the cartridge directly. Several parameters affecting the extraction efficiency were investigated, including the type of elution, washing solution, and sample pH. Under the optimized conditions, excellent limits of quantitation for the target analytes were found to be 0.15–1.18 ng/mL, and the average recoveries of the analytes at two spiked levels for real‐sample analysis ranged from 69.8 to 106.2% with RSDs less than 7.3%. Furthermore, the graphene‐based cartridges exhibited superior reusability for juice sample analysis. The proposed method is sensitive, simple, and cost saving, and provides a detection platform for the monitoring of pesticide residues.     

High-performance liquid chromatographic determination of furfural and hydroxymethylfurfural in apple juices and concentrates

Summary A rapid and sensitive method for determining 2-furaldehyde (FUR) and 5-hydroxymethyl-2-furaldehyde (HMF) in apple juices and juice concentrates has been developed. The method for FUR and HMF involves the solid-liquid extraction of the juice by using a C-18 cartridge prior to reversed-phase separation with detection at 280 nm. The mobile phase was acetonitrile-water (8/92, v/v) at a flow rate of 1.0 ml/min. Recoveries from apple juices and juice concentrates spiked at different levels ranged from 94.1 to 104.0 (FUR) and 94.5 to 100.5 (HMF). The quantification limit for both, FUR and HMF, was 5 ppb.     

Liquid chromatographic method for quantitation of patulin at 10 ng/mL in apple-based products intended for infants: interlaboratory study

An interlaboratory trial for the determination of patulin in apple juice and fruit puree was conducted, involving 17 participants representing a cross section of industry, official food control, and research facilities. Mean recoveries reported ranged from 74 (10 ng/g) to 62% (25 ng/g) for apple juice and from 72 (25 ng/g) to 74% (10 ng/g) for fruit puree. Based on results for spiked samples (blind pairs at 2 levels), as well as naturally contaminated samples (blind pairs at 3 levels), the relative standard deviation for repeatability (RSDr) in juice ranged from 8.0 to 14.3% and in puree from 3.5 to 9.3%. The relative standard deviation for reproducibility (RSD(R)) in juice ranged from 19.8 to 39.5% and in puree from 12.5 to 35.2%, reflecting HORRAT values from 0.6 to 1.0 for juice and 0.4 to 0.9 for puree. The method showed acceptable within-laboratory and between-laboratory precision for each matrix, as required by current European legislation.     

Liquid chromatography/mass spectrometric determination of patulin in apple juice using atmospheric pressure photoionization

This paper describes a comparison between atmospheric pressure chemical ionization (APCI) and the recently introduced atmospheric pressure photoionization (APPI) technique for the liquid chromatography/mass spectrometric (LC/MS) determination of patulin in clear apple juice. A column switching technique for on-line extraction of clear apple juice was developed. The parameters investigated for the optimization of APPI were the ion source parameters fragmentor voltage, capillary voltage, and vaporizer temperature, and also mobile phase composition and flow rate. Furthermore, chemical noise and signal suppression of analyte signals due to sample matrix interference were investigated for both APCI and APPI. The results indicated that APPI provides lower chemical noise and signal suppression in comparison with APCI. The linear range for patulin in apple juice (correlation coefficient >0.999) was 0.2-100 ng mL(-1). Mean recoveries of patulin in three apple juices ranged from 94.5 to 103.2%, and the limit of detection (S/N = 3), repeatability and reproducibility were 1.03-1.50 ng mL(-1), 3.9-5.1% and 7.3-8.2%, respectively. The total analysis time was 10.0 min.     

Determination of the carbon-13 content of sugars and pulp from fruit juices by isotope-ratio mass spectrometry (internal reference method). A European interlaboratory comparison

An interlaboratory comparison by ¹³C-isotope ratio mass spectrometry was organised by working group No. 1 of the European Commission of Standardisation (CEN/TC 174) in order to define the repeatability (r) and the reproducibility (R) of the ¹³C determination in sugars and pulp, isolated from the same fruit juice. Six unlabelled juices were analysed in 19 laboratories in the European Union, Australia and the USA. Three samples were authentic juices (orange, grapefruit, pineapple) and the remaining samples were the same juices with an addition of sugar at 15 g l⁻¹ (orange, pineapple) or 11.8 g l⁻¹ (grapefruit). The sugar was prepared by mixing equal amounts of beet and cane sucroses. The different laboratories used the same experimental protocol under different conditions (operator, conversion system for CO₂ preparation, mass spectrometer). The results for the sugars (r = 0.27%., R = 0.82%.) were comparable to those from an earlier ring test (r = 0.30%., R = 0.70%.), while the results for the pulp presented higher interlaboratory variability (r = 0.38%., R = 1.89%.) possibly due to experimental difficulties. Nevertheless, the RSD_R for free sugars ranged 0.9–3.2% and for pulp, 0.9–9.3%. On the basis of the Horwitz equation and taking account of the concentration of the isotopes measured, an RSD_R of 7–8% might have been expected. The experimentally determined RSD_R values were less than twice the theoretically expected values, which is a criterion of an acceptable method. The results were therefore considered to be acceptable by specialists in the field of isotopic analysis, and the method applied was found to improve the sensitivity of the ¹³C determination for the detection of sugar addition in fruit juices.     

Gas chromatographic-mass spectrometric characterization of flavanones in citrus and grape juices.

A method for the characterization of flavanones in fruit juices, involving solvent extraction, hydrolysis to the corresponding aglycones, trimethylsilylation and combined gas chromatography-mass spectrometry, is reported. The application of the method is demonstrated for the analysis of orange, lemon, grapefruit and grape juices.