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Author(s): Jyoti Goswami, Thakur Vikram Singh, Chhaya Bhatt, Deepak Kumar Sahu, Kalpana Wani, Ajay Kumar Sahu, Prashant Mundeja, Manish Kumar Rai*, Joyec Rai

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Address: School of Studies in Chemistry, Pt. Ravishankar Shukla University Raipur (Chhattisgarh), 492010, India
Chhattisgarh Council of Science and Technology, Saddu Rd Mowa, Raipur (Chhattisgarh), 492014, India

Published In:   Volume - 33,      Issue - 1,     Year - 2020

Cite this article:
Goswami et al. (2020). An Extractive Spectrophotometric Method for the Determination of Pymetrozine in Various Ecological Samples of Bilaspur District (C.G.). Journal of Ravishankar University (Part-B: Science), 33(1), pp. 65-72.

An Extractive Spectrophotometric Method for the Determination of Pymetrozine in Various Ecological Samples of Bilaspur District (C.G.)

Jyoti Goswami1*, Thakur Vikram Singh1, Chhaya Bhatt1, Deepak Kumar Sahu1, Kalpana Wani1, Ajay Kumar Sahu1, Prashant Mundeja1, Manish Kumar Rai1, Joyec Rai2

1School of Studies in Chemistry, Pt. Ravishankar Shukla University Raipur (Chhattisgarh), 492010, India

2Chhattisgarh Council of Science and Technology, Saddu Rd Mowa, Raipur (Chhattisgarh), 492014, India.


*Corresponding author:,

[Received: 13 December 2019; Revised version: 26 September 2020; Accepted: 28 September 2020]

Abstract: Pesticides are designated mixture of substances for extenuating and destroying any group of pests such as insects and vegetation. Pymetrozine [6-methyl-4-[(E)-pyridin-3-ylmethylideneamino]-2, 5-dihydro-1, 2, 4-triazin-3-one] is a pyridine azomethine compound which represents a class of insecticide. A Spectrophotometric method has been developed on the modified Fujiwara reaction for the determination of pymetrozine. Pymetrozine directly reacts with chloroform the 45-500C turns out the violet color in the formation of Schiff’s Base (Glutaconic Aldehyde). In the present reaction, the violet color obtained is discharged with a few drops of acetic acid followed by the addition of p- nitro aniline reagent which gives yellow color dyes. The absorption maxima of yellow-colored dye measured at 430 nm. Beer’s law is obeyed over the range of 1 to 10 µg per 10 ml of pymetrozine. The molar absorptivity of the colored system is 1.38×105 L mol-1 and Sandall’s sensitivity is 1.0×10-3µg cm-2. This method is free from the interference of other interfering and can be successfully applied for the determination of various ecological samples.

Keyword: Fujiwara Reaction, Pymetrozine, Chloroform, Schiff’s Base, Ecological Sample.


Pymetrozine (C10H12N5O), pyridine azomethine speaks to another substance class of bug sprays with a momentous selectivity for plant-sucking creepy crawlies, for example, aphids, whiteflies, and planthoppers, because of its foundational Action [Neuen et al., 2013]. Pymetrozine rapidly affects the feeding behavior of insect pests. Pymetrozine may take effect through the nervous system, with a unique mode of action that differs from other well-known insecticides [Ausborn et al 2005; Hong et al 2011]. Pymetrozine is probably not going to sully groundwater at the prescribed application in Owing to its high effectiveness, low poisonousness, high selectivity, and ecological amicability, Pymetrozine has pull in far reaching enthusiasm as a pesticide. Pymetrozine 50 WG is the bug spray from the pyridine azomethines bunch with a one of a kind method of activity that keeps the creepy crawly from embeddings the stylet into the plant tissue [Cui et al., 2013; Deekshita and Ramarao 2018].

The present investigation is carried out to evaluate the efficacy of Pymetrozine along with different insecticides against Ecological field conditions [Boina et al., 2011; Qiong et al., 2012]. Consequently, pymetrozine buildups may prompt potential wellbeing injury. A few investigation strategies have been produced for deposits of Pymetrozine in various frameworks, for example, differential spectrophotometric method molar absorptivity and sandal’s sensitivity observed [Jixin et al., 2018; Zhang et al., 2015]. A simple, sensitive, and selective method were obeying beers lambert law for quantification of Pymetrozine was developed to study the dissipation behavior and final residue of Pymetrozine in various fields in the ecosystem. However, the current knowledge of the behavior of Pymetrozine in Ecological systems is lacking. It is important to explore the sorption-desorption processes of Pymetrozine and factors that modulate its samples [Jansen et al 2011; Mingxing et al 2016] LC-MS analysis of pesticides in soya grain samples, [Pizzuti et al., 2007]. Pymetrozine main metabolites, CGA128632, GS23199, and CGA266591 in Chinese kale using liquid chromatography with tandem triple quadrupole mass spectrometry LC-MS [Gong et al., 2018]. The mechanisms responsible for contaminant removal were elucidated at different pH, temperature, contact time and initial ions concentration using sorption kinetics and isotherms [Cui et al., 2013]. LC-MS/MS determination with comparison by two other methods of extraction as described [Abdelselam et al., 2006], HPLC-UV with liquid-liquid partition cleanup, Pre-treatment [Zhang et al., 2007]. FTIR microspectroscopy is combined with principal component-discriminant function analysis [Baker et al., 2018]. Gold nanoparticles colorimetric detection of organophosphorus pesticide [Bala et al., 2016], ultrasound-assisted microextraction of Carbaryl pesticides from water sample [Bazrafshan et al., 2017], residue analysis of Pymetrozine [Gong et al., 2018], detection of organophosphate pesticide in rewal samples [Yola et al., 2019], physical, chemical and biological Food analysis, food safety and human health analysis [shingh et al., 2019], argent need for a new concept of agriculture [Stamati et al., 2016] were analyzed.



A Systronic UV-Vis spectrophotometer model – visiscan167 with coordinated silica cell was utilized for all ghostly estimations. A Systronic pH meter model – 335 was used for pH assessment.


All reagents (HCl and NaOH) used were of Anala. R. grade and Double Distilled water were utilized all through.

Standard solution of pymetrozine

Supplied by Godrej Agrovet Ltd, a stock solution of 1mg/ml solution of pymetrozine is prepared in water. The working standard solution is prepared by appropriate dilution of the stock solution.


Supplied by Thermo Fisher Scientific India Pvt. Ltd, A 5.0 mol L-1 aqueous solution is used.

P-nitro aniline reagent

Supplied by Loba Chemie Pvt. Ltd, 0.001M solution of p-nitro aniline with ethanol is prepared.


An aliquot containing 1 to 12 μg of Pymetrozine was taken1 ml in 25 ml of the calibrated test tube. The solution of the test tube was evaporated off up to 0.5 ml on a water bath. To this 1ml of chloroform followed by 2ml of 5 M sodium hydroxide were added and the test tube was kept in a boiling water bath for 45-500C temperature. The violet-colored dye obtained was cooled in ice-cold water and then de-colorized with few drops of glacial acetic acid. Then 1ml of p-nitro-aniline reagent and 10 M HCl 0.5 ml was added and the solution was kept for 10 minutes for full-color development. The volume of the test tube was made up to 10 ml with distilled water and the absorbance of the yellow-colored dye was measured at 430 nm against a reagent blank.(Sharma et al.,2003)

Chemical Reaction (color reaction)

The reaction place three steps in the first step Pymetrozine reacts with chloroform in an alkaline medium to obtain violet color (I), in the second step on the addition of glacial acetic acid, the violet color disappear of formation of Schiff;’s base of glutaconic aldehydes (II) and is converted into glutaconic aldehydes (II), which form a yellow-colored dye (III) with p-nitro aniline reagent in the third step (Scheme 1).

Results and Discussion

Spectral Characteristics: The p-nitro aniline dye formed in the proposed reaction shows maximum absorption at 430 nm (Figure 2). All ghostly estimations were completed against double refined water as the reagent clear indicated irrelevant intake at this frequency [Eldridge et al., 2018]. The shading structure complies with Beer's law in the range of 1 to 100 μgml-1 of pymetrozine last arrangement at 430 nm (Figure 3 to 6). Table 1is shows the reproducibility of Method Pymetrozine in days investigation, Table 2 is presented Optical characteristics and Statistical Data of the Regression Equations for the Reaction, Table 3 is presented The effect of Foreign Species, and Table 4 is presented the recovery of pymetrozine in Various Ecological Samples [Wani et al., 2016].


Table 1. Reproducibility of Method

No. of Days

Absorbance at 430nm

















Standard deviation


Relative Standard Deviation


The concentration of Pymetrozine used was 7μg /10mL


Table 2. Optical characteristics and Statistical Data of the Regression Equations for the Reaction

S. No.


Values for the reaction


Lemda Max nm



Beer,s law limit, µg mL-1

1 to 12


Detection limits, µg mL-1



Quantification limit µg mL-1



Molar Absorptivity, L mol-1 cm-1



Sandell,s sensitivity, µg cm-2



Regression equation, y = b+a

0.045X + 0.252








Standard Deviation



Relation Standard deviation (%)



Correlation coefficient(r2)






Table 3. Effect of Foreign Species

Foreign Species & Ione

Tolerance limit* μg in 10 mL

















Concentration of Pymetrozine 7 μg / 10 mL.

*The amount causing an error of ± 2% in absorbance value.

Table 4. Recovery of pymetrozine in Various Ecological Samples


(μg mL-1)

(μg mL-1)

Total found
(μg mL-1)


(% ± R.S.D.)






96.6 ± 0.0244






99.43 ± 0.041






96.26 ± 0.056






98.26 ± 0.042






98.9 ± 0.042






99.56 ± 0.041






99.8 ± 0.041






99.43 ± 0.027






99.73 ± 0.041






99.3 ± 0.15

***Mean of three replicate analyses

*Water sample 5 mL

**Amount of Soil Sample 5 gm


Optimization of condition

The precision of the method was checked by the replicate analysis of a working standard solution containing 7μg of Pymetrozine in a 10 ml final solution over for 7 days. The standard deviation and relative standard deviation were found to be ± 0.002 and 0.39 % respectively. The molar absorptivity, Sandell’s sensitivity, correlation coefficient, regression equation intercept, slope, LOQ, and LOD were found to be Table 3.

Effect of Chloroform: 1ml of chloroform was sufficient for complete development. A higher or lower amount of chloroform decreased the absorbance value.

Effect of Temperature: it was found that 2-3 minutes in a water bath a temperature range 45 to 500C was sufficient for the complete color development further heating and increasing the temperature of the solution decreases the absorbance value.

Effect of Reagent: it was found that 1 mL of p-nitro aniline was sufficient for complete color development. On increasing the amount of p-nitro aniline the absorbance value decreases.

Effect of Foreign Species: effect of various pollutants and pesticides on the determination of Pymetrozine was studied. To a sample containing 7 µg of Pymetrozine known of foreign species and pesticide were added. The Pymetrozine was analyzed as described above the method was found to be free from the interference of other pesticides and pollutants are normally given Table-3[Parween et al., 2017].


Determination of Pymetrozine in water sample: 100ml of water taken and fortified with a known amount of Pymetrozine add and kept for 6 hours, Pymetrozine was then extracted in ethanol. Ethanol was evaporated off and Pymetrozine was determined by the proposed as well as reported Pymetrozine completely soluble in ethanol. The recuperations are appeared in the Table 4.

Determination of Pymetrozine in soil sample: 5 gm of soil taken and fortified with known amount of Pymetrozine   and 10 mL double distilled water add and kept for 6 hours, and then these samples are filtered through Whatman No. 40 filter paper. Pymetrozine was determined by the proposed as well as reported [Mahmood et al., 2015].  The recuperations are appeared in the Table 4.

Determination of Pymetrozine in vegetables, Fruits Grains, and soil: Potato, Carrot, Tomato, Gherkin, Apple, Bean, Rice, and Lemon were weighed (5 gm), crushed, and spiked with a known amount of Pymetrozine. After 6 hours Pymetrozine was extracted in ethanol. Ethanol was evaporated off and Pymetrozine was determined by the proposed as well as reported method the recoveries are shown in Table 4. The recoveries range from 96 – 99% by the present method.



The proposed method is fast, straight-forward, touchy, and the reagent depicted here is delicate, and specific for experiments of Pymetrozine in different environmental samples, and strategy is more particular than other detailed methods. It can effectively apply for the assurance of Pymetrozine in water, soil, vegetable Fruits, and Grain analysis. Check the recuperations, known measures of Pymetrozine were added to different samples of vegetables, products of the soil tests, and afterward broke down by the proposed strategy (Table 4).


Authors are appreciative of the head, school of studies in science Pt. Ravishankar Shukla University, Director General, Chhattisgarh Council of Science and innovation and, University Gant Commission for giving research laboratory facility and award separately.


Abdelselam, S., John S., Wilkie, A., Kevin, N., (2006). Synthesis of Some Hydrazones Derivatives Structurally Related to the Insecticide Pymetrozine. Journal of Chemistry 49, 927-930.

Ausborn,  J., Wolf, H., Mader, W., Kayser,  H.,  (2005). The insecticide pymetrozine selectively affects chordotonal mechanoreceptors, Journal of Experimental Biology The Journal of Experimental Biology 208, 4451-4466

Baker, M.J.,  Gazi, E., Brown, M.D.,  Shanks, J.H.,  Gardner,P., Clarke,N.W.,(2008). FTIR-based spectroscopic analysis in the identification of clinically aggressive prostate cancer, British Journal of Cancer ,99, 1859 – 1866

Bala, R., Sharma R.K., Wangoo, N., (2016). Development of gold nanoparticles-based aptasensor for the colorimetric detection of organophosphorus pesticide phorate. Analytical and Bioanalytical Chemistry 408, 333–338.

Bazrafshan, A.A., Ghaedi, M., Rafiee, Z., Hajatib, S., Ostovan, A., (2017). Nano-sized molecularly imprinted polymer for selective ultrasound-assisted microextraction of pesticide Carbaryl from water samples: Spectrophotometric determination. Journal of Colloid and Interface Science Volume 498, 313-322

Boina D.  R., Youn Y., Folimonov S. and Stelinski L L (2011) Effects of pymetrozine, an antifeedant of Hemiptera, on Asian citrus psyllid, Diaphorina citri, feeding behavior, survival, and transmission of Candidatus Liberibacter asiaticu, Society of Chemical Industry Pest Management Science  67: 146–155

Cui, L,Yan J., Quan, G., Ding, C., Chen, T., Hussain, Q., (2013) Adsorption Behavior of Pymetrozine by Four Kinds of Biochar from Aqueous Solution, Adsorption Science & Technology 31 No. 6

Deekshita K and Ramarao C. V. (2018)  Pymetrozine: A Pyridine Azomethine insecticide for management of rice brown planthopper in India  Chemical Science  Review Letters, 7(25), 335-339

Eldridge, B., F., (2008). Pesticide Application And Safety Training For Applicators Of Public Health Pesticides, Applicators May Be Obtained From The California Department Of Public Health, (916) 552-9730. 

Gong,J., Zheng K., Yang, G., Zhao, S., Zhang, K., Hu., D., (2018). Determination, residue analysis, risk assessment and processing factor of pymetrozine and its metabolites in Chinese kale under field conditions, Food Additives & Contaminants: Part A

Hong,  J., Lee C., Lim J., (2011).  Comparison of Analytical Methods and Residue Patterns of Pymetrozine in Aster scaber, Bull Environ Contam Toxicol 87:649

Jansen, J.P., Defrance, T., Warnier, A.M., (2011). Side effects of flonicamide and pymetrozine on five aphid natural enemy, species Bio Control 56:759–770

Jixin, Yu., Genbo, Xu., Wei, Li., Shiyu, J., Ting, Yu.,  Jiashou, Li., Zhongjie, Li., Tanglin, Z., (2018). Pymetrozine: A Pyridine Azomethine insecticide for management of rice Intetnational Journal of Environmental Research. Public Health 15, 984;

Mahmood, S., Imad, I.R.S., Shazadi, K., Gul Z.A., Khalid, Hakeem, R., (2015). Effects  Of  Pesticides  On  Environment, Springer International Publishing Switzerland (Eds.), Plant, Soil And Microbes, DOI 10.1007/978-3-319-27455-3_13,253-269.

Mingxing, G.A.O., Yingying, LI., Hong, Y., Yucheng, G.U., (2016). Sorption and desorption of pymetrozine on six Chinese soils,  Frontiers. Environmentsal. Science.and Engineering 10(1): 1–10

 Nauen, R., Vonta,s J., Kaussmanna, M., Wolfel, K., (2013) Pymetrozine is hydroxylated by CYP6CM1, a cytochrome P450 conferring neonicotinoid resistance in Bemisia tabaci1'Society of Chemical,  Industry Pest Management Science 69: 457–461* and T. Sherwood I

Parween,T., Jain, S., (2017). Interface Between Pesticide Chemistry And Plant Physiology, Ecophysiology Of Pesticide, Elsevier, ISBN 978-0-12-817614-6,

Qiong, R., Hua, Xu.Y., Chen, L.U.O., Hong,  Z., Jones, C.M., Greg, J., Devine, G.J., Gorman, K., Denholm, I.,  (2012). Characterisation of Neonicotinoid and Pymetrozine Resistance in Strains of Bemisia tabaci from China, Journal of Integrative Agriculture 11(2): 321-326

Singh, P. K., Singh, R. P., Singh, P., Singh, R. L. (2019). Food Hazards: Physical, Chemical, And Biological. Food Safety And Human Health, 15–65. DOI:10.1016/B978-0-12-816333-7.00002-3.

Stamati, P., N., Maipas, S., Kotampasi, C., Stamatis, P., Hens, L., (2016). Chemical Pesticides and Human Health: The Urgent Need for a New Concept in Agriculture, Front Public Health. 4, 148. doi: 10.3389/fpubh.2016.00148.

Tsujimoto, K., Sanad, S., Masaya, M., (2016). A new method for monitoring the susceptibility of the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae), to pymetrozine by combining topical application and measurement of offspring number,  Journal of Entomology Zooogy studies 51, 155–160

Wani K., Nirmal M., Patel, V., Khatoon, R., Rai, MK., Rai, J(2016) Determination of Carbendazim in Environmental sample with Iron (III) and 1, 10 phenanthroline as Reagents. Asian journal of Chemistryvalume 29, 161-165

Yola, M.L., (2019). Electrochemical activity enhancement of monodisperse boron nitride quantum dots on graphene oxide: Its application for simultaneous detection of organophosphate pesticides in real samples. Journal of Molecular Liquids. 277, 50-57

Zhang, Y.Li., Zhang, Li., Peng, Xu., Jianzhong, Li., Wang, H., (2015). Dissipation and residue of pymetrozine in rice field ecosystem, Environatal Monitoring Assessment  187: 78

 Zhou Q Du F., Shi Y., Liu W, Liu D and Chen G (2018) Isolation and characterization of a pymetrozine grading strain Pseudomonas sp. BYT-1  journal of chemical research 42, 434–438



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