Analysis of Pesticide Residues in Curry Leaves and Red Gram in Tirupati Region, Chittoor by Gas Chromatography Vineela, Ramya Kuber B*
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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue -6, Nov-Dec- 2017 http://dx.doi.org/10.22161/ijeab/2.6.32 ISSN: 2456-1878 Analysis of Pesticide Residues in Curry Leaves and Red Gram in Tirupati Region, Chittoor by Gas Chromatography Vineela, Ramya Kuber B* Institute of Pharmaceutical Technology, Sri Padmavati Mahila Viswavidyalayam, Tirupati, Andhra Pradesh, India Abstract— Medicine is food and food is medicine” is the sprayed on crops. So the present work is aimed to determine best way to describe on how the ailments were cured by the pesticide residue levels in Curry leaves and Red gram in using the plants during the ancient period of time. The Tirupati region, Chittoor by using Gas Chromatography. “Magical plant of Indian Spice” (Murraya koenigii) has Pesticides which are determined in this article are served human kind not only as food enhancer but also serve chlorpyrifos, profenofos, triazophos. Chlorpyrifos is a broad as village or folk medication to cure many disorders, the spectrum organophosphate pesticide, kills the insects and tribal communities has used many parts of the worms on crops, animals, buildings(Mackay et al.,2014). Murrayakoenigiito cure them diseases. Pigeonpea Profenofos is a non-systemic broad spectrum, foliar [Cajanuscajan(L.) Millsp.]Pigeon pea (Arhar) commonly insecticide and acaricide. It shows excellent translaminar known as Red gram or tur is a very old crop in India. The action and effective against chewing and sucking insects, term ‘Pigeon pea’ was coined in Barbados . The present mites on cotton, rice, maize, sugar beet, soybeans, vetables, study is to determine the pesticide residue levels in Curry tobacco, oil seeds etc., (Madhulika kushwaha et al. 2016). leaf (Murraya koenigii) and Red gram (Peagion Pea) in Triazophos is a broad spectrum, non systematic contact Tirupati , Chittoor region. organosphosphorus pesticides used to control pests, insects, Keywords— Curry leaves, Gas Chromatogrpahy, Pesticide acarids, some nematodes in okra, cotton, maize, rice Standards, Red Gram. paddies etc.,(Aungpdradit et al. 2007).Pesticide residues in curry leafs was determined in different markets of Andhra I. INTRODUCTION Pradesh & Telangana by using LC-MS/MS.(Priyadarshini Murraya koenigii, commonly known as curry leaf or et al., 2017) karivepaku or Kari patta in Indian dialects, belonging to Family Rutaceae which represent more than 150 genera and II. MATERIALS AND METHODS: 1600 species (Gabriel Charles and Vincent Onuegbu, 2014). Market samples of curry leaf and Red gram were collected Fresh leaves, dried leaf powder, and essential oil are widely from local market Tirupati region Chittoor. Samples were used for flavoring soups, curries, fish and meat dishes, eggs extracted for pesticide residues following the validated dishes, traditional curry powder blends, seasoning and ready QuEChERS method to give best results. to use other food preparations (Gupta et al.,1971) . It is 2.1 Sample extraction: called as ‘Magical plant of Indian spice’. It is not only used Curry leaf samples were analyzed for pesticide residues as food enhancing purpose but also shows medication for following the AOAC official method (QuEChERS) which many diseases(Surbhisinghal and meenakshibatt, 2016). is the best method in the laboratory. The samples were The essential oil is also utilized by soap and cosmetic collected from local Tirupati market in polythene bag. aromatherapy Industry. Peagion Pea which belongs to the Curry leaves was homogenized separately with robot coupe family Fabaceae, is commonly known as Red gram ,in Blixer. 5g of sample was weighed and taken in 50 ml Andhra pradesh we called as Kandi pappu, other names centrifuge tube and 10ml distilled water, 15 ml acetonitrile like Tur, arhar, Dhal etc. This is very old crop and major was added to sample tube. The sample was homogenized at cultivated crop in India (Gowda et al., 2011). These crops 14000-15000rpm for 2-3 min. 6g sodium chloride was were contaminated with pesticides, not only these two, added to sample, mixed thoroughly by shaking gently almost all crops are contaminated with pesticide which are followed by centrifugation for 3min at 2500-3000rpm to www.ijeab.com Page | 3033 International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue -6, Nov-Dec- 2017 http://dx.doi.org/10.22161/ijeab/2.6.32 ISSN: 2456-1878 separate the organic layer. The top organic layer of about residues is crucial for proper risk assessment of human 9ml was taken into the 15ml centrifuge tube and added with exposure through food. 1.4g magnesium sulphate to remove the moisture content Compared with other available methods, the QuEchERS and added 1g PSA sorbent (for dispersive solid phase d- method is believe to give the best results. SPE cleanup), and 0.05g of GCB (Graphitized Carbon 4.1 GC determination of pesticide standards: Black) shaken gently followed by centrifuge for 2min at The pesticide residue was identified by comparing its 2500rpm. The sample tube was vortexed for 30 sec then retention time (Rt). The quantitative determination was followed by centrifuge for 5 min at 2500-3000rpm. 2ml carried out with help of a calibration curve. A good linearity supernant layer was transferred into 10ml tube for was established by a correlation coefficient (R2) value evaporation using water bath and reconstitute with 1ml of n- (0.97). hexane. Now pour it in GC vials and inject it in GC having Correlation coefficient is a statistical tool used to measure FPD detector. Same procedure were followed for Red gram. the degree of this type relationship and here a high Weigh individual reference standard around 15mg in 25ml correlation value (A value very close to1.0) indicates a high volumetric flask. Dissolve in n-hexane for standards to be level linear relationship between the concentration of used on GC-FPD and adjust to 500ppm removing required standards and peak area. For quantification an external quantity and making up to 25ml. The working standards are calibration curve with four different concentration matrix prepared by serial dilutions from stock solution i.e 0.1, 0.25, matching were made. 0.5, 0.75, 1ppm by suitable solvent n-hexane and used as The standards are dissolved in n-hexane it was injected in standard check in analysis linearity studies(Swarupa Rani et GC as an blank. In blank chromatogram we observed only al., 2016). one peak i.e solvent peak as shown in Fig.1 III. INSTRUMENT ANALYSIS The chromatographic system was SHIMADZU AOC-20S 2010 plus, equipped with an Auto sampler and EB1 Column 0.25mm×0.25µm, 30m length. The determination of pesticide residue analysis was carried out by following conditions - Injection volume 1µl, Run time 20min, Detector Flame photometric detector(FPD), Injection 0 temperature 260 c, Column flow 6ml/min, column Fig.1: The blank chromatogram with solvent peak 0 programming 60 c, Column EB1, Detector, temperature 2800c the run time will change for standards. IV. RESULTS AND DISCUSSION The pesticide residue levels are calculated by following formula: Residues(µg/g) : height or area of sample µl of sample injected x x height or area of the sample µl of the standard injected conc. of standard / weight of sample x final volume Fig.2: The Standard chromatograms of Chlorpyrifos - equation (1) Pesticides are applied to Vegetables, leafy vegetables, fruits Peak Retention Area Height Name and various crops at various stages of cultivation and for time protection against a range of pests during harvest before 1 17.242 338391 649767 Chlorpyr they become available to consumer. 8 ifos To ensure the safety of food for consumers and regulate Tota 338391 649767 international trade, legislations such as European Union l 8 directives has established maximum residue limits for pesticides in food stuff. Thorough monitoring of pesticide 4.1.1 Linearity for the Chlorpyrifos: www.ijeab.com Page | 3034 International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue -6, Nov-Dec- 2017 http://dx.doi.org/10.22161/ijeab/2.6.32 ISSN: 2456-1878 The linearity was established by analysing standard Peak Retention Area Height Name solutions of the insecticides at five concentrations. The time concentrations were 0.1, 0.25, 0.5, 0.75, 1µg/ml. Graphs 1 20.445 1985369 243885 Profenofos were constructed from the GC chromatograms based on the Total 198569 243885 average peak area of the signal response versus the concentration plot of the analytes. The parameters such as 4.1.2 Linearity for Profenofos: the slope of the regression line, y-intercept and the The linearity of the method was determined at five correlation coefficient were evaluated. concentration levels ranging from 0.1-1 ppm for profenofos. The regression line equation for Profenofos was Y=2E+06Xand the regression coefficient value was 0.994 Y- AXIS respectively. 4000000 The regression data for the calibration curve showed good linear relationship over a concentration range 0.1-1ppm for 3000000 y = 4E+06x y- axis Profenofos with respect to peak area. 2000000 R² = 0.9769 1000000 Linear (y- 0 axis) 2500000 Y-AXIS 0 1 2 2000000 y = 2E+06x Fig.3: Linearity of chloropyrifos R² = 0.9941 1500000 Y-AXIS Table : 1 1000000 Sl no. Concentration(µg/ml) Area Linear (Y- 1 0.1 256874 500000 AXIS) 2 0.25 1022717 0 3 0.5 1572715 0 0.5 1 1.5 4 0.75 2910537 Fig.5: Linearity of profenofos 5 1 3383918 6 Slope 4E Table : 2 7 Y-intercept 06x+4E Sl no Concetration(µl/ml) Area 8 Correlation coefficient 0.976 1 0.1 175354 2 0.25 570827 The linearity of the method was determined at five 3 0.5 997618 concentration levels ranging from 0.1-1ppm for 4 0.75 1600129 Chlorpyrifos. The regression line equation for Chlorpyrifos 5 1 1985369 was Y=4E+06X and the regression coefficient value was 6 Slope 2E 0.967 respectively.