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Optimization of a Multi-Residue Method for 101 Pesticides in Green Tea

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Optimization of a Multi-Residue Method for 101 Pesticides in Green Tea Revista Brasileira de Farmacognosia 26 (2016) 401–407 ww w.elsevier.com/locate/bjp Original Article Optimization of a multi-residue method for 101 pesticides in green tea leaves using gas chromatography–tandem mass spectrometry a,b,∗ a,b a,b a,b Xue Hou , ShaoRong Lei , LingAn Guo , ShiTing Qiu a Center of Analysis and Testing, Sichuan Academy of Agricultural Sciences, Chengdu, China b Laboratory of Quality and Safety Risk Assessment for Agro-products (Chengdu), Ministry of Agriculture, Chengdu, China a r a b s t r a c t t i c l e i n f o Article history: A method for analysis of 101 pesticide residues in tea leaves was developed and validated for the first time. Received 13 January 2016 Pure acetonitrile was used as extraction solvent rather than acetonitrile after matrix hydration based on Accepted 2 March 2016 the amount of co-extracts and recoveries performance. During clean-up procedure, primary-secondary Available online 13 April 2016 amine/graphitized carbon black (500 mg) was selected, which exhibited outstanding properties in clean- up capabilities and recoveries of pesticides comparing to primary-secondary amine/graphitized carbon Keywords: black (250 mg), NH2-Carbon and TPT absorbents. The method was validated employing gas chromatogra- Tea leaves −1 phy coupled to tandem mass spectrometry at the spiked concentration levels of 0.050 and 0.100 mg kg . Pesticide residues For most of the targeted pesticides, the percent recoveries range from 70 to 120%, with relative standard SPE 2 GC–MS/MS deviations <20%. The linear correlation coefficients (r ) were higher than 0.99 at concentration levels −1 −1 PSA/GCB of 0.025–0.250 mg kg . Limits of quantification ranged from 1.1 to 25.3 ␮g kg for all pesticides. The developed method was successfully applied to the determination of pesticides in tea leaf samples. © 2016 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Introduction and solid-phase extraction (SPE) (Huang et al., 2009). Apart from these more common approaches, during the analysis of some pes- Tea, the dried leaves of the plant of Camellia sinensis (L.) Kuntze, ticide residues, head-space solid-phase micro-extraction (SPME) Theaceae (Yang and Landau, 2000), is an aromatic beverage that (Schurek et al., 2008) and stir bar sorptive extraction (SBSE) (Li consumed worldwide. Due to its properties of antioxidant, antimi- et al., 2012) were also successfully applied. Another famous way crobial, anticarcinogenic and anti-inflammatory (de Mejia et al., for the detection of multi-residues is known as QuEChERS (quick, 2009), tea has attracted great attention. However, tea farming easy, cheap, effective, rugged and safe), which involves MeCN is sensitive to many kinds of diseases, pests and weeds, which extraction and purification with dispersed solid-phase extraction causes the widely use of pesticides. Until now, more than 300 (d-SPE) (Anastassiades et al., 2003a,b; Zhang et al., 2010). The main kinds of pesticide residues in tea have been reported (Pang et al., concept of QuEChERS for sample processing strategy is to use dif- 2011). Pesticides may cause potential health risk to consumers and ferent type of absorbents on the basis of the component of matrix, impose great pressure on the environment (Jaggi et al., 2001), thus for instance: primary-secondary amine (PSA), C18 silica, Florisil many countries have established maximum residue limits (MRL) and graphitized carbon black (GCB). Hayward et al. have devel- for many pesticides, such as European Community (EC) no. 42/2000 oped a multi-residues determination method in botanical dietary and (EC) no. 1881/2006 (Li et al., 2013). supplements using PSA/GCB as clean-up materials. Whereas, in Meanwhile, tremendous efforts have been performed in order this study, several SPE materials were compared depending on to develop analytical methods for pesticides determination in tea. clean-up capabilities and recoveries of pesticides (Hayward et al., Generally, the preparation method of multi-residues analysis is 2013). carried out in a sequence of several steps, including extraction For the detection techniques, at the beginning, gas chromatog- with solvent, purification and detection. For the sample clean-up raphy (GC) and high performance liquid chromatography (HPLC) procedure, there are several efficient ways, such as gel perme- with variety of detectors were widely employed to detect pes- ation chromatography (GPC) (Pang et al., 2006; Huang et al., 2007) ticide residues. The most commonly used detectors including electron capture detector (ECD) (Xia et al., 2008), flame photometric detector (FPD) (Moinfar and Hosseini, 2009), nitrogen phospho- ∗ rus detector (NPD) (Oh, 2007) for GC and fluorescence detector Corresponding author. E-mail: [email protected] (X. Hou). (FLD) (Wu et al., 2009), diode array detection (DAD) (Sharma et al., http://dx.doi.org/10.1016/j.bjp.2016.03.007 0102-695X/© 2016 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/). 402 X. Hou et al. / Revista Brasileira de Farmacognosia 26 (2016) 401–407 2008) for HPLC. Owing to better selection, sensitivity and higher Sample preparation and clean-up throughput in the detection of multi-residues, chromatography coupled to MS (Chen et al., 2012; Zhao et al., 2012) and tan- Sample preparation dem MS (MS/MS) (Zhao et al., 2012; Cajkaa et al., 2012) have Tea leaves were collected from organic tea plantation located in been widely used nowadays. These techniques enable the analy- Sichuan province in China during March 2014, which was identified sis of multi-residues simultaneously with lower LODs and higher as Camellia sinensis (L.) Kuntze cv. Mengshan 9 by Prof. Yun Wang accuracy. (Tea Research Institute, Sichuan Academy of Agricultural Sciences). Unfortunately, although control of pesticides in tea leaves may The tea leaf samples were comminuted with dry ice and homoge- increase the safety of tea from the source, for such detection nized, and then 50 g subsamples were kept frozen until spiking or method, little research has been done until now. In this study, we analysis. describe for the first time the method of 101 residues in tea leaves Tea leaf subsamples (5 g) were weighed in polypropylene cen- based on the application of GC–MS/MS combined with optimized trifuge tubes (50 ml) and 20 ml MeCN were added. The solution was extraction and SPE clean-up procedures. Moreover, the proposed then vortexed for 1 min. QuEChERS extraction bag with 4 g anhy- method has been successfully applied to the determination of these drous MgSO4, 1 g NaCl, 1 g tri-sodium citrate dehydrate (tri-Na) pesticides in real samples. and disodium hydrogencitrate sesquihydrate (di-Na) was added, and the tube was vortexed immediately to prevent coagulation of ◦ MgSO4 for 1 min. After centrifugation (1210 g, 10 min, −10 C), 5 ml Experimental of the upper acetonitrile layer was transferred and purified with SPE. Material and chemicals SPE procedure HPLC-grade MeCN, acetone, toluene and hexane were provided SPE cartridges were preconditioned with 5 ml MeCN–toluene by Tedia (Fairfield, OH, USA). Prepacked QuEChERS extraction bags (3:1, v/v). Then concentrated extract was introduced into the car- with 4 g MgSO , 1 g sodium chloride (NaCl), 1 g tri-sodium citrate 4 tridge. For all experiments, MeCN–toluene (3:1, v/v) was used as dehydrate (tri-Na) and disodium hydrogencitrate sesquihydrate eluting solvent. The eluents were collected and then concentrated (di-Na) were purchased from Agilent (Palo Alto, CA, USA). Standard to dryness using vacuum concentration. Finally, the residue was pesticides (Table 1) with purities ranging from 95 to 99% were redissolved in 2.5 ml mixture of n-hexane and acetone (9 + 1; v/v) supplied by Sigma–Aldrich (Madrid, Spain) and Dr. Ehrenstorfer for GC–MS/MS analysis. (Augsburg, Germany). Stock solutions of mixture pesticides were ◦ prepared in acetone and stored in freezer (−18 C). The working Gravimetric determination of co-extracts solutions were prepared daily. For SPE, PSA/GCB (500 mg) cartridges, PSA/GCB (250 mg) car- For sample extraction procedure, three types of 5 ml of crude tridges, and NH2-Carbon cartridges were supplied by Agilent (Palo MeCN extracts obtained by pure MeCN, MeCN after matrix hydra- Alto, CA, USA). TPT cartridges were obtained from Agela Technolo- tion using 5 ml and 10 ml water were evaporated separately until gies (Tianjin, China). dryness with a weak nitrogen stream, and the residues were gravi- metrically determined by analytical balance. Average amounts of five replicates were evaluated (Fig. 1), the error bars (standard devi- Equipment ations) of the weights for each group were also showed. For clean-up procedure, 5 ml of crude MeCN extracts and 5 ml of A Vortex (IKA, Germany), a vacuum distillation apparatus (IKA, MeCN extracts purified through four types of SPE were evaluated Germany) and a centrifuge (Xingke, China) were used for preparing according to gravimetric method described above (Fig. 3). the samples. GC–MS/MS system (Varian, Inc., USA) was employed, equipped with a Varian 450 gas chromatograph (equipped with a CP Method performance 8400 autosampler and a 1079 injector) and a 320 triple quadrupole MS. A VF-5 MS fused silica capillary column of 30 m × 0.25 mm The precision and accuracy of the method was tested with I.D. and 0.25 ␮m was used (Agilent, Inc., USA). Helium (purity ≥ −1 spiked tea leaves. Recoveries were determined for five repli- 99.999%) was used as a carrier gas at a flow rate of 1.2 ml min . −1 cates at two spiking concentrations (0.050 and 0.100 mg kg ).
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