Detection and Confirmation of Transfluthrin in Viscera- a Case Study

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International Journal of Forensic Sciences MEDWIN PUBLISHERS ISSN: 2573-1734 Committed to Create Value for researchers Detection and Confirmation of Transfluthrin in Viscera- A Case Study

Chandra S1*, Yadav S1, Kori S2, Anjana1 and Shinde LP3 1Chemistry Division, Forensic Science Laboratory, India Case Report 2 Chemistry, biochemistry & forensic science, Amity school of applied sciences, Amity University Volume 5 Issue 1 Haryana, India Received Date: February 04, 2020 3 Department of Chemistry, NES Science College, Nanded, India Published Date: March 04, 2020 DOI: 10.23880/ijfsc-16000178 *Corresponding author: Subhash Chandra, Chemistry Division, Forensic Science Laboratory, Rohini, New Delhi, India, Tel: +919049847414; Email: [email protected]

Abstract

Mosquito repellents such as vaporizers, mats, coils and creams are being used in the household to reduce the risk of dengue, chickungunya and other infectious diseases. Nowadays liquid mosquito repellent is very common in the society due to easy availability, low cost and easy to use it remains available in most of the houses but its misuse in suicide/homicides cases can- not be ruled out. Toxicity of repellents results mainly in central nervous system (CNS) symptoms like headache, dizziness, drowsiness, status epilepticus and respiratory failure. A case of unknown poisoning was reported in 62 year old deceased and an empty repellants bottle was found at scene of crime. In this study various solvent systems have been tried to detect

transfluthrin in biological material by using thin layer chromatography (TLC). It was observed that Benzene: Hexane (1:1) solvent system found very close Rf value with reference to standard transfluthrin in comparison to other solvent systems. The confirmation compound was carried out by FTIR and Gas Chromatography Mass Spectrometry (GC-MS) methods. The developed solvent system was found the quick, easy and cost effective for detection of transfluthrin in biological material. It found in the brain. has also been found that the concentration of the transfluthrin prevalent in the in tissues of liver and kidney, lowest levels was

Keywords:

Transfluthrin; Viscera; Case study; TLC; FTIR; GC-MS Introduction The analytical methods of insecticides have been improved in last few years by advance and sophisticated techniques In India, the patterns of ingestion poison have been [6]. Amarnath, et al. have reviewed several techniques for changed. The earlier days common poison like opium, the qualitative as well as quantitative analysis of insecticides arsenic, oleander and dhatura, etc. were used for poisoning ranging from conventional chromatographic methods purpose [1]. Nowadays it has been replaced by insecticides like organophosphorous, organochlorine, carbamate and better, efficient, fast and result oriented method that can pyrethroids [2-4]. Pyrethroids are effective pest control beto theutilized modern in future GC-MS work methods, of analysis in order [7]. toThe suggest method a chemical with low mammalian toxicity are increasingly of approach to an analysis of biological material for the used in Indian agriculture and household purpose. Due to presence of drugs depends very much on the type of quarrel in the families are routine matter, but poisoning material provided. Dhingra, et al. have detected ranitidine in visceral material by thin layer chromatography methods. of insecticides in body fluids is the choice of research from veryby transfluthrin long time before in houseto know is the unusual impact thingof its on [4,5]. living Analysis beings. H12Cl2F4O2

Transfluthrin (C15 ; M.W: 371.15 g/mol

Detection and Confirmation of Transfluthrin in Viscera- A Case Study Int J Forens Sci 2 International Journal of Forensic Sciences

solvent systems and analytical techniques were reported for dichlorovinyl)-2,2-dimethyl-cyclopropanecarboxylate) is aIUPAC: colourless crystal, 2,3,5,6-tetrafluorobenzyl-(1R,S)-trans-3-(2,2- this pyrethroid pesticide with a broad been tried to develop various solvent systems for analysis spectrum, acting by contact, inhalation and repellent by its ofdetection pesticide and residuesconfirmation in biological of transfluthrin material. pyrethroid. For routine It has strong deadly capability, and is effective to prevent and cure toxicological analysis thin layer chromatographic method hygienic and storage pests (Figure 1). It has swift deadly was developed for analysis of biological and non-biological effect to pests of diptera such as mosquitoes, and very good sample. residual effect to cockroaches and bedbugs. It can be used to produce coil, aerosol preparation and mats [8]. Case History A 62 year old male patient was admitted in the emergency ward of All India Institute of Medical Sciences (AIIMS), New Delhi, in unconscious state by his relatives. The relatives have informed that the patient had taken chicken curry with his family at night and went to sleep and it was observed by his relatives in unconscious state at 3.00AM. The patient had a history of thyroid and high blood pressure as informed by the relatives. The doctors have observed that pinpoint pupils, increased respiratory rate, breathing

Figure 1 saliva and sweat and muscle twitching, convulsions. The livedifficulty saving and treatment bronchospasm. was given, There the were patient excess was secretions, not found : Molecular Structure of Transfluthrin. Uses of Transfluthrin deteriorated and patient was declared dead at 11.00 AM by thefit to doctors. give statement The postmortem during entire was conducted treatment. the The viscera condition was send to Forensic Science Laboratory in sealed condition for mosquito repellent and vaporizer, its solutions designed for toxicological examinations. aerosolsTransfluthrin and in the widelyformulation used of as liquid mats, pesticides. coils and It can liquid be purchased at different types of retail outlets [9]. poisoning by organophosphorus compounds but none of the organophosphorusThe findings of postmortem poison couldindicated be towardsdetected suspected however, persistency, may cause symptoms of poisoning including nervousness,Transfluthrin anxiety, is atremor, fast acting convulsions, pyrethroid skin withallergies, low literature indicated that incidents of accidental poisoning a transfluthrin a pyrethroid was detected. The review of are known, but symptomatic treatment with antihistamines maysneezing, help runningto control nose any and allergies. irritation. It has No beenspecific reported antidotes by of transfluthrin is rare but this is the first fatal case of Panwar, et al. the use of liquid mosquito repellent vaporizer transfluthrinExperimental poisoning was detected. is common in setting of infections like malaria and dengue [10]. These products contain derivatives of pyrethrins as Chemicals and Reagents mosquito repellent and kerosene as the solvent. Pyrethrins have poor dermal absorption and rapid metabolism. Popular ether, Ethyl Acetate, n-Hexane, Methanol, Anhydrous Sodium Acetic Acid, Acetone, Benzene, Chloroform, Diethyl the present case. Toxicity results mainly in central nervous brands use 0.88% of the chemical that was transfluthrin in Merck analytical grade. system (CNS) symptoms like headache, dizziness, drowsiness, Sulphate, Rhodamine B, Sodium Carbonate were used E status epilepticus and respiratory failure. These chemicals act by prolonging the opening of sodium channels leading Apparatus/Instruments of the nervous system. Cardiac dysfunction has also been pipettes, sprayer, sample applicator, TLC plates pre-coated described,to increased but influx lung of injury sodium is not ions commonly and thus reportedhyper-excitation [11]. Separating funnel, volumetric flask, conical flask,

withPreparation silica gel 60 F256 of Solvent (Merck), System FTIR and GC-MS. in past history but none were fatal and patients survived by the symptomaticThe poisoning treatment. cases due The to transfluthrin fatal poisoning have case been is rare.seen The use of liquid-liquid extraction technique for the It has been revealed from the literature survey that various

extraction of transfluthrin has been carried out and further

Sci 2020, 5(1): 000178. Chandra S, et al. Detection and Confirmation of Transfluthrin in Viscera- A Case Study. Int J Forens Copyright© Chandra S, et al. 3 International Journal of Forensic Sciences subjected to different solvent systems and spraying reagent. samples extracted from the viscera were spot on a TLC plate along with the standard solution of insecticides. the compound by TLC methods. Development of TLC Plate: The spot plates were placed The solvent system has been developed for identification of inside the chromatographic chamber, which was previously Preparation of Reagents saturated with solvent system used as mobile phase and was allowed to run, later on the plate was removed from the TLC Spraying Reagent: Spray 1: chamber and air-dried. The different ratios of used solvent systems were as follows. Spray 2: 0.2g of sodium carbonate 0.05g in distilled of Rhodamine water. B was dissolved in 50 ml methanol. Solvent System: then air dried and sprayed with spraying reagent 2. Hexane: Acetone (8:2) Sprayed the developed plate first with spraying reagent 1 Various solvent systems has been used as Preparation of Standard Solution: Standard stock solutions Hexane : Ethyl Acetate (8:2) (1000 µg/mL) were prepared individually by dissolving Hexane :Chloroform :Acetic Acid(9:5:0.5)

VisualizationBenzene : Hexane of (1:1) TLC Plates: The developed plates were anddifferent Pyrethrin. pyrethroids in n-hexane (HPLC Grade). The sprayed with the spraying reagents and their Rf values were standards used are Transfluthrin, Cypermethrin, Allethrin Extraction of Transfluthrin from Viscera calculated and compared with the standard transfluthrin. In a portion of 100 gmbiological tissues marked andThe their developed Rf values plates were were calculated first observed and compared under with 256 nmthe of UV light and then sprayed with the spraying reagents added 10 gm of anhydrous ammonium sulfate powder for ‘1A’(containing stomach and small intestine), in conical flask deproteinization purpose, after shaking pH was adjusted standard (STD) transfluthrin (Figure 2). to 7.0 by the addition of ammonical buffer solution. The exhibits were extracted with 100 ml of ethyl acetate in a separating funnel. The ethyl acetate extract was transferred to an evaporating dish and the aqueous phase re-extracted through anhydruous sodium sulphate to remove moisture. Thetwice extract with 25 was ml ofevaporated ethyl acetate to anddryness. the solution The residue was passed then dissolved in 2.0ml of n-Hexane. The same process was

(containing portion of liver, spleen and kidney) and ‘1D’ (containingcarried out withportion the of 100 brain) gm of(Clarke’s1969). biological tissue marked ‘1B’

The extraction was done from blood sample marked ‘1C’ sulphuric acid and solution of sodium tungstate and pH was adjustedby taking to10ml 7.0 of by it inadding conical ammonical flask and deproteinizing buffer solution. it withThe Figure 2: extraction analyte was done using ethyl acetate and residue was dissolved by n-Hexane. Acetone::8:2 TLC plate(b)Hexane: of Transfluthrin Trichloroethane: spot of extract Acetic held under UV-light at various solvent system (a)Hexane: Acetate::8:2. Acid::9:5:5 (c)Benzene: Hexane::1:1 (d)Hexane: Ethyl The detection of the transfluthrin was carried FTIR Analysis: The moisture free extracted sample was out from the extracted exhibits i.e. ‘1A’, ‘1B’ and ‘1C’ and ‘1D’ by TLC methods and confirmation was done analyzed using Fourier Transform Infrared Spectroscopy using Preparation instrumental of TLC techniques System (FTIR and GC-MS). The sample (S-1) extracted from the viscera was comparedalong with the standard spectra solutions of the both of standardtransfluthrin. and sample. The sample spotted on a TLC plate along with the standard solution of and standard taken and prepared pellets with KBr and insecticides. GC-MS Analysis : The GC-MS analysis has been carried out Spotting of Sample and Standard on TLC Plate: The for confirmation of the transfluthrin. GC-MS System; Carrier gas : Helium; GC injector temperature: 275° C; Oven : 50°

thin layer chromatography with better distinction from other pyrethroid (Cypermethrin, Allethrin and Pyrethrin) as earlier InjectionC(1min); 25°mode:split C/min toless; 150° Injection C(0 min); volume:1µL; 3° C/ min Total to 200° run conventional solvent systems were giving almost similar time:C; 8° C/min40 min.; to 280°Mass C (Shimadzu),(10 min); post Polarity: run : 320° EI+, CElectron (5min);

Rf value making the detection process difficult. The solvent Solventenergy (ev):delay:4 70; min. MS temperature: 200° C (Source); Transfer andsystem sample Hexane: (S-1). BenzeneThis solvent (1:1) found was reliable found and to deliver suitable best for Column:line temperature : 275° C; Trap (µA): 200; Repellor (v): 8.0; results as close Rf values obtained for standard transfluthrin

MS: DB5 MS; Polarity: EI +, Electron energy (ev): 70; the detection transfluthrin using thin layer chromatographic TheMS temperature: auto sampler 200° was Cused (Source); to inject Transfer 1.0 µl lineof each temperature: extracted inmethods. viscera The has method delivered developed best results for analysis as compared of transfluthrin to other 275° C; Trap (µA): 200; Repellor (v):8.0, Solvent delay:4 min. solventby using system TLC with used Hexane: in this Benzene study. The (1:1) TLC as methodsolvent systemis very cheap, less time consuming and can be performed in any sampleResults into the and GC-MS. Discussion The retention time was 13.06 min.

laboratory efficiently. repellent vapoziser and has been seen in non-fatal poisoningTransfluthrin cases [12]. is aIn pyrethroid this study useda fatal in case liquid of suspected mosquito elutionThe time extracted and Total samples Ion Chromatogram were further confirmed(TIC)obtained for organophosphorous poisoning was revealed by the transfluthrin by FTIR and GC-MS technique [14,15]. The spectra was analyzed for presence of prominent peaks and was compared with that of transfluthrin standard. The FTIR investigating officer but after analysis finally detected materialTransfluthrin [13]. insecticide belong to the pyrethroids group compared with spectra of the transfluthrin standard. The by using TLC, GC-MS and FTIR techniques in the biological deceasedanalysis of [16] both (Figure the instrumental 3-6). techniques confirmed the presence of transfluthrin in the Biological materials of the

An attempt has been made to detect transfluthrin using

Figure 3: FTIR Spectra of Standard Transfuthrin.

Figure 4: FTIR Spectra of Sample (std).

Figure 5

: TIC of Transfluthrin Sample.

Figure 6: MS Spectra of Sample and Standard.

Conclusion and support for conducting this research work. References in viscera of deceased which may responsible for cause 1. In this case study, Transfluthrin has been detected ( suspected poisoning by organophosphorus compounds Djilani A, Legseir B, Soulimani R, Dicko A, Younos C butof death.none Theof the findings organophosphorus of postmortem poison indicated was detected towards 2006) New extraction technique for alkaloids. J Braz 2. Chem Soc 17(3): 518-520. The review of literature indicated that incidents of accidental however, transfluthrin a pyrethroid pesticide was detected. ofRay Organophosphate AK, Ghosh MC (2006) & Carbamate Aquatic Compounds.Toxicity of Carbamates Academic and Organophosphates. In: Ramesh CG (Ed.), Toxicology poisoning of transfluthrin is rare but this is the first fatal 3. Fukuto TR (1990) Mechanism of action of effectivecase of transfluthrin and safe. poisoning was detected. The developed Press, pp: 657-672. solvent system for detection of transfluthrin is quick, cheap, organophosphorus and carbamate insecticides. Environ

Acknowledgement 4. Health Perspect 87: 245-254. and Mr Sri Narain, Head of Division (Chemistry), Forensic Peter JV, John G, Cherian AM (1996) Pyrethroid poisoning. Authors are thankful to Mrs. Deepa Verma, Director Science Laboratory, Rohini, Delhi, for their encouragement 5. J Assoc Physicians India 44(5): 343-344.

Scott JG (1988) Pyrethroid insecticides. ISI Atlas Sci

11. Cardiac Conduction Disturbance Due To Prallethrin ( 6. Pharmacol 2(2): 125-128. PyrethroidBhaskar EM, ) MoorthyPoisoning. S, GaneshwalaJ Med Toxicol G, 6(1): Abraham 27-30. G (2010) determination of 22 organophosphorous pesticides inF Musshoff,human blood H Junker, using B headspace Madea (2002) solid-phase Simple 12. microextraction and gas chromatography with mass spectrometric detection. J Chromatogr Sci 40(1): 29-34. Andrés M, Lorenz LM, Mbeleya E, Moore SJ (2015) AnophelesModified mosquitoarabiensis mosquitoes landing boxesunder dispensingsimulated 7. transfluthrin provide effective protection against

Mishra A, Shukla SK, Gupta AK (2015) Analysis of 13. outdoorNunes MJ, conditions Camões inMF, a semi-field Fournier system.J (1997) Malar Analysis J 14(1). of Insecticides in Body Fluids: A Review. J Chromatograph organophosphorus, organochlorine and pyrethroid 8. SeparatPauluhn Techniq J, Ozaki 5(4): K ( 1-3. insecticides in medicinal plants. Chromatographia 44(9-

Regul Toxicol Pharmacol2015) 71(1): Transfluthrin: 78-92. Comparative efficacy and toxicity of reference and generic versions. 14. 10): 505-513. 9. Jeyalakshmi T, Shanmugasundaram R, Kannadasan J, Material.Clarke EGC J Med (1970) Chem Isolation 13(2): 338.and Identification of Drugs in Pharmaceuticals, Body Fluids and Post Mortem variousGeetha S,room Hilda sizes S, et against al. (2014) Culex Efficacy quinquefasciatus of a commercial Say. J 15. (2013) Analysis of chlorpyrifos Entomolliquid vaporiser Zool Stud ( Transfluthrin 2(3): 220-224. 0 . 88 % ( w / v )) under 6(4):Neti 466-472. N, Zakkula V 10. degradation by Kocuria sp . using GC and FTIR. Curr Biot An association with pyrethroid poisoning. Indian J Crit 16. (1999) Organophosphorus and Carbamate CarePanwar Med M, 17(2): Usha G,119-120. Kumath M (2013) Status epilepticus: 287-294.Glaser LC Pesticides. U.S. Geological Survey, Reston, VA, USA, pp: