ICP-MS ile Lucilia Sericata (Meigen, 1826) [Diptera: Calliphoridae] Larvalarından Arsenik ve Kadmiyum İntoksikasyon Olgularının Tahmini

Abstract

Amaç: Bu çalışmada, kontamine karaciğer dokusu ile beslenen Lucilia sericata (Meigen, 1826) larvalarında arsenik (As) ve kadmiyum (Cd) tespiti için İndüktif Eşleşmiş Plazma-Kütle Spektrometrisi (ICP-MS) temelli bir analitik yöntemin geliştirilmesi ve valide edilmesi ile beslenme ortamı ve larvalardaki element konsantrasyonları arasındaki ilişkinin değerlendirilmesi amaçlanmıştır.

Yöntem: Farklı As ve Cd konsantrasyonları ile kontamine edilen tavuk karaciğeri temelli beslenme ortamlarına yaklaşık 100 L. sericata yumurtası inoküle edilmiş ve larvalar üçüncü evreye ulaşıncaya kadar üç gün inkübe edilmiştir. Larvalar nitrik asit ve hidrojen peroksit ile parçalanarak ICP-MS ile analiz edilmiştir. Kör larvalardan matriks uyumlu kalibrasyon eğrileri hazırlanmış; yöntem performansı doğrusallık, LOD, LOQ, geri kazanım ile gün içi ve günler arası yeniden üretilebilirlik açısından değerlendirilmiştir.

Bulgular: Matriks uyumlu kalibrasyon eğrileri her iki element için 1–20 ng/mL aralığında doğrusal bulunmuştur. Geri kazanım değerleri As için %94,33–107,38, Cd için %97,98–108,16 arasında belirlenmiştir. Gün içi bağıl standart sapma değerleri As için %3,46–8,51, Cd için %2,22–10,36 bulunmuştur. As ve Cd’ye maruz kalan larvaların ortalama uzunluklarının kör gruba göre daha düşük olduğu gözlenmiştir. Beslenme ortamı ve larvalardaki element konsantrasyonları arasındaki korelasyon katsayıları As için 0,677, Cd için 0,902 olarak hesaplanmıştır.

Sonuç: Geliştirilen ICP-MS yöntemi, L. sericata larvalarında As ve Cd’nin doğru ve tekrarlanabilir şekilde tespit edilmesini sağlamıştır. Bulgular, larval element analizinin özellikle geleneksel biyolojik matrikslerin bulunmadığı şüpheli post-mortem zehirlenme olgularında adli araştırmalara katkı sağlayabilecek toksikolojik bilgi sunabileceğini göstermektedir.

Keywords

• Adli entomoloji
• entomotoksikoloji
• Lucilia sericata
• ICP-MS
• arsenik
• kadmiyum

Supporting Institution

İstanbul Üniversitesi-Cerrahpaşa Bilimsel Araştırma Projeleri Birimi, Türkiye

Project Number

FYL-2020-3450

Ethical Statement

Bu çalışmada deneysel model olarak omurgasız bir tür olan Lucilia sericata larvaları kullanılmıştır. İstanbul Üniversitesi-Cerrahpaşa Hayvan Deneyleri Yerel Etik Kurulu yönergelerine göre omurgasız türlerle yapılan çalışmalar etik kurul onayı gerektirmemektedir. Beslenme ortamı olarak kullanılan tavuk karaciğeri yerel bir perakende marketten ticari olarak temin edilmiş olup, bu çalışma için özel olarak hiçbir hayvan sakrifiye edilmemiştir. Çalışma insan katılımcı içermediğinden bilgilendirilmiş gönüllü onamı uygulanabilir değildir.

Thanks

Bu çalışma, FYL-2020-3450 proje numarası ile İstanbul Üniversitesi-Cerrahpaşa Bilimsel Araştırma Projeleri Birimi tarafından desteklenmiştir.

Prediction of Arsenic and Cadmium Intoxication Cases from Lucilia Sericata (Meigen, 1826) [Diptera: Calliphoridae] Larvae by ICP-MS

Abstract

Aim: This study aimed to develop and validate an Inductively Coupled Plasma-Mass Spectrometry (ICP-MS)-based analytical method for detecting arsenic (As) and cadmium (Cd) in Lucilia sericata (Meigen, 1826) larvae fed on contaminated liver tissue and to evaluate the relationship between element concentrations in the feeding media and larvae.

Method: Chicken liver-based media spiked with different As and Cd concentrations were inoculated with approximately 100 L. sericata eggs and incubated for three days until the third instar stage. Larvae were digested with nitric acid and hydrogen peroxide, then analyzed by ICP-MS. Matrix-matched calibration curves were prepared from blank larvae, and method performance was assessed for linearity, limit of detection (LOD), limit of quantification (LOQ), recovery, and intra- and inter-day reproducibility.

Results: Matrix-matched calibration curves were linear in the range of 1–20 ng/mL for both elements. Recovery values ranged from 94.33% to 107.38% for As and from 97.98% to 108.16% for Cd. Intra-day relative standard deviation values were 3.46–8.51% for As and 2.22–10.36% for Cd. Larvae exposed to As and Cd showed shorter average lengths than the blank group. The correlation coefficients between element concentrations in the feeding media and larvae were 0.677 for As and 0.902 for Cd.

Conclusion: The developed ICP-MS method provided accurate and reproducible detection of As and Cd in L. sericata larvae. These findings suggest that larval elemental analysis may provide useful toxicological information in forensic investigations, particularly when conventional biological matrices are unavailable in suspected post-mortem poisoning cases.

Keywords

• Forensic entomology
• entomotoxicology
• Lucilia sericata
• ICP-MS
• arsenic
• cadmium

Supporting Institution

Scientific Research Projects Unit of Istanbul University-Cerrahpaşa, Türkiye

Project Number

FYL-2020-3450

Ethical Statement

The experimental model used in this study involved larvae of Lucilia sericata, an invertebrate species. According to the guidelines of the Istanbul University-Cerrahpasa Animal Experiments Local Ethics Committee, studies involving invertebrates do not require ethical approval. Chicken liver used as feeding media was commercially obtained from a local retail market, and no animals were sacrificed specifically for this study. Since the study did not involve human participants, informed consent was not applicable.

Thanks

This study was supported by the Scientific Research Projects Unit of Istanbul University-Cerrahpasa, Türkiye, under project number FYL-2020-3450.

References

1. 1. Catts EP, Goff ML. Forensic entomology in criminal investigations. Annu Rev Entomol. 1992;37(1):253-272. doi: 10.1146/annurev.en.37.010192.001345
2. 2. Joseph I, Mathew D, Sathyan P, Vargheese G. The use of ınsects in forensic ınvestigations: an overview on the scope of forensic entomology. J Forensic Dent Sci. 2011;3(2):89.
3. 3. Amendt J, Campobasso CP, Gaudry E, Reiter C, LeBlanc HN, JR. Hall M. Best practice in forensic entomology—standards and guidelines. Int J Legal Med. 2007;121(2):90-104.
4. 4. Benecke M. Six Forensic Entomology Cases: Description and Commentary. J Forensic Sci. 1998;43(4):797-805. doi: 10.1520/JFS14309J
5. 5. Sohal RS, Lamb RE. Intracellular deposition of metals in the midgut of the adult housefly, Musca domestica. J Insect Physiol. 1977;23(11-12):1349-1354.
6. 6. Sohal RS, Lamb RE. Storage-excretion of metallic cations in the adult housefly, Musca domestica. J Insect Physiol. 1979;25(2):119-124. doi: 10.1016/0022-1910(79)90089-1
7. 7. Beyer J, Enos W, Stajić M. Drug ıdentification through analysis of maggots. J Forensic Sci. 1980;25(2):411-412. doi: 10.1520/JFS12147J
8. 8. Wilson Z, Hubbard S, Pounder DJ. Drug analysis in fly larvae. Am J Forensic Med Pathol. 1993;14(2):118-120. doi: 10.1097/00000433-199306000-00002

9. 9. Sadler DW, Fuke C, Court F, Pounder DJ. Drug accumulation and elimination in Calliphora vicina larvae. Forensic Sci Int. 1995;71(3):191-197. doi: 10.1016/0379-0738(94)01663-1
10. 10. Levine B, Golle M, Smialek JE. An Unusual Drug Death Involving Maggots: Am J Forensic Med Pathol. 2000;21(1):59-61. doi: 10.1097/00000433-200003000-00010
11. 11. Rezende F, Alonso MA, Souza CM, Thyssen PJ, Linhares AX. Developmental rates of immatures of three Chrysomya species (Diptera: Calliphoridae) under the effect of methylphenidate hydrochloride, phenobarbital, and methylphenidate hydrochloride associated with phenobarbital. Parasitol Res. 2014;113(5):1897-1907.
12. 12. Kintz P, Tracqui A, Mangin P. Analysis of opiates in fly larvae sampled on a putrefied cadaver. J Forensic Sci Soc. 1994;34(2):95-97. doi: 10.1016/S0015-7368(94)72890-5
13. 13. Bourel B, Fleurisse L, Hédouin V, et al. Immunohistochemical contribution to the study of morphine metabolism in calliphoridae larvae and ımplications in forensic entomotoxicology. J Forensic Sci. 2001;46(3):596-599. doi: 10.1520/JFS15009J
14. 14. Kharbouche H, Augsburger M, Cherix D, et al. Codeine accumulation and elimination in larvae, pupae, and imago of the blowfly Lucilia sericata and effects on its development. Int J Legal Med. 2008;122(3):205-211. doi: 10.1007/s00414-007-0217-z
15. 15. Nuorteva P, Nuorteva SL. The fate of mercury in sarcosaprophagous flies and in insects eating them. Ambio. 1982;11(1):34-37.
16. 16. Nolte K, Pinder R, Lord W. Insect larvae used to detect cocaine poisoning in a decomposed Body. J Forensic Sci. 1992;37(4):1179-1185. doi: 10.1520/JFS13304J
17. 17. Amendt J, Richards CS, Campobasso CP, Zehner R, Hall MJR. Forensic entomology: applications and limitations. Forensic Sci Med Pathol. 2011;7(4):379-392.
18. 18. Roeterdink EM, Dadour IR, Watling RJ. Extraction of gunshot residues from the larvae of the forensically important blowfly Calliphora dubia (Macquart) (Diptera: Calliphoridae). Int J Legal Med. 2004;118(2):63-70. doi: 10.1007/s00414-003-0408-1
19. 19. Rashid RA, Arifuddin N, Ahmad NW. Blowfly, Chrysomya megacephala as an alternative specimen in determination of gunshot residue. In: 2012 IEEE Symposium on Business, Engineering and Industrial Applications. IEEE; 2012:542-547.
20. 20. Cil M, Mercan S, Cakan H. Prediction of the number of firearm shots by using Lucilia sericata (Diptera: Calliphoridae) larvae with an ınductively coupled plasma -mass spectrometry (ICP-MS) technique. Entomol News. 2025;132(3).
21. 21. Boulkenafet F, Bouhayene S, Benzazia S, et al. Heavy metal bioaccumulation in rabbit organs and Chrysomya albiceps (Wiedemann, 1819) larvae: ımplications for forensic entomology. Indian J Anim Res. 2024;(Of). doi:10.18805/IJAR.BF-1745
22. 22. Anderson G. Insect succession on carrion and its relationship to determining time of death. In: Forensic Entomology: The Utility of Arthropods in Legal Investigations. 2000:143-175.
23. 23. Dadour IR, Cook DF, Fissioli JN, Bailey WJ. Forensic entomology: application, education and research in Western Australia. Forensic Sci Int. 2001;120(1-2):48-52.
24. 24. Kökdener M, Yılmaz AF. The effects of gunshot residue components (Pb, Ba, and Sb) on the life history traits of Lucilia sericata (Diptera: Calliphoridae). Lysyk T, ed. J Med Entomol. 2021;58(6):2130-2137. doi: 10.1093/jme/tjab123
25. 25. Malejko J, Deoniziak K, Tomczuk M, Długokencka J, Godlewska-Żyłkiewicz B. Puparial cases as toxicological indicators: bioaccumulation of cadmium and thallium in the forensically important blowfly Lucilia sericata. Front Chem. 2020;8:586067.
26. 26. Hughes MF, Beck BD, Chen Y, Lewis AS, Thomas DJ. Arsenic exposure and toxicology: a historical perspective. Toxicol Sci. 2011;123(2):305-332. doi: 10.1093/toxsci/kfr184
27. 27. Vahidnia A, Van Der Voet GB, De Wolff FA. Arsenic neurotoxicity — A review. Hum Exp Toxicol. 2007;26(10):823-832. doi: 10.1177/0960327107084539
28. 28. Chen QY, Costa M. Arsenic: A global environmental challenge. Annu Rev Pharmacol Toxicol. 2021;61(1):47-63. doi: 10.1146/annurev-pharmtox-030220-013418
29. 29. Bissen M, Frimmel FH. Arsenic — a Review. Part I: occurrence, toxicity, speciation, mobility. Acta Hydrochim Hydrobiol. 2003;31(1):9-18. doi: 10.1002/aheh.200390025
30. 30. Smedley PL, Kinniburgh DG. A review of the source, behaviour and distribution of arsenic in natural waters. Appl Geochem. 2002;17(5):517-68.
31. 31. Marchiset-Ferlay N, Savanovitch C, Sauvant-Rochat MP. What is the best biomarker to assess arsenic exposure via drinking water? Environ Int. 2012;39(1):150-171. Kile ML, Hoffman E, Hsueh YM, et al. Variability in Biomarkers of Arsenic Exposure and Metabolism in Adults over Time. Environ Health Perspect. 2009;117(3):455-460.
32. 32. Loffredo CA, Aposhian HV, Cebrian ME, Yamauchi H, Silbergeld EK. Variability in human metabolism of arsenic. Environ Res. 2003;92(2):85-91.
33. 33. Meija J, Coplen TB, Berglund M, et al. Atomic weights of the elements 2013 (IUPAC Technical Report). Pure Appl Chem. 2016;88(3):265-291. doi: 10.1515/pac-2015-0305
34. 34. Norman NC, ed. Chemistry of Arsenic, Antimony and Bismuth. 1. ed. Blackie Academic & Professional; 1998.
35. 35. Satarug S. Cadmium sources and toxicity. Toxics. 2019;7(2):25.
36. 36. Satarug S. Dietary Cadmium Intake and Its Effects on Kidneys. Toxics. 2018;6(1):15.
37. 37. Kumar S, Sharma A. Cadmium toxicity: effects on human reproduction and fertility. Rev Environ Health. 2019;34(4):327-338. doi: 10.1515/reveh-2019-0016
38. 38. Salmela SS, Vuori E, Huunan-Seppälä A, Kilpiö JO, Sumuvuori H. Body burden of cadmium in man at low level of exposure. Sci Total Environ. 1983;27(1):89-95.
39. 39. Vuori E, Huunan-Seppälä A, Kilpiö JO, Salmela SS. Biologically active metals in human tissues. II. The effect of age on the concentration of cadmium in aorta, heart, kidney, liver, lung, pancreas and skeletal muscle. Scand J Work Environ Health. 1979;5(1):16-22.
40. 40. Kjellström T. Exposure and accumulation of cadmium in populations from Japan, the United States, and Sweden. Environ Health Perspect. 1979;28:169-197.
41. 41. Subhan F, Khan A, Wahid F, Shehzad A, Jan AU. Determination of Optimal Toxic Concentration and Accumulation of Cadmium in Broiler Chicks. Toxicol Res. 2011;27(3):143-147. doi: 10.5487/TR.2011.27.3.143
42. 42. Hsu CW, Weng CH, Lin-Tan DT, et al. Association of Urinary Cadmium with Mortality in Patients at a Coronary Care Unit. Kokubo Y, ed. PLoS One. 2016;11(1):e0146173.
43. 43. Ammann AA. Inductively coupled plasma mass spectrometry (ICP MS): a versatile tool. J Mass Spectrom. 2007;42(4):419-427. doi: 10.1002/jms.1206
44. 44. Wilschefski SC, Baxter MR. Inductively Coupled Plasma Mass Spectrometry: Introduction to Analytical Aspects. Clin Biochem Rev. 2019;40(3):115-133.
45. 45. Costa RA, Dos Santos NA, Corrêa TSM, et al. Detection of Pb, Ba, and Sb in Cadaveric Maggots and Pupae by ICP‐MS. J Forensic Sci. 2020;65(6):2188-2193.
46. 46. Roe A, Higley LG. Development modeling of Lucilia sericata (Diptera: Calliphoridae). PeerJ. 2015;3:e803. doi: 10.7717/peerj.803
47. 47. Bugelli V, Campobasso CP, Verhoff MA, Amendt J. Effects of different storage and measuring methods on larval length values for the blow flies (Diptera: Calliphoridae) Lucilia sericata and Calliphora vicina. Sci Justice. 2017;57(3):159-164.
48. 48. Bambaradeniya TB, Magni PA, Dadour IR. Morphological changes of larvae and pupae of Lucilia sericata (Diptera: Calliphoridae) reared at two temperatures and on three food types. Byrd J, ed. J Med Entomol. 2024;61(3):521-529. doi: 10.1093/jme/tjae013
49. 49. Altunsoy F, Başaran A. Effects of Thallium on the Larval Development of Lucilia sericata Meigen 1826 and PMI Estimation. Anadolu Univ J Sci Technol C Life Sci Biotechnol. 2011;1(2):103-112.