The accurate and reliable HPLC-UV based method for detecting the active ingredient hexaflumuron in some plant protection products.

Year 2025, Volume: 7 Issue: 2, 162 - 167, 31.05.2025

Ismael Mohamed

https://doi.org/10.51435/turkjac.1651242

Abstract

The main objective of this study was to present a new selective, sensitive, and validated RP-HPLC analytical method for measuring the concentrations of hexaflumuron (HFM) which is a well-known active ingredients that is widely used as an insecticide to protect crops, especially fruits and vegetables. Quantification was carried out using a reversed phase HPLC system that was equipped with a UV detector. The development of the novel method was performed on reversed-phase C-18 (stainless steel, 5 µm, 250 × 4.6 mm) column at constant temperature 30 ºC. The mobile phase consists of acetonitrile and distilled water in volumetric ratio 85:15, a flow rate of 1 mL/min, and the detection wavelength at 220 nm. Retention time of separation at 3.84 min. The method was validated by testing specificity, linearity, precision, recovery, LOD, LOQ, and accuracy according to the CIPAC (Collaborative International Pesticides Analytical Council) and comply with the guidelines of SANCO/3030/99 rev.5. guidelines. The method revealed an acceptable linearity regression R2 (0.9974). The method was found to be accurate from concentration levels 50–250 µg/mL with high accuracy (99.3-100.7%). The method’s validation results make it suitable for use as a standardization tool in the evaluation of Emulsion Concentrate formulations containing this active ingredient.

Keywords

Hexaflumuron , HPLC Analysis , Method validation , Precision , Accuracy

Supporting Institution

KZ for Pesticides and Chemicals Company

Thanks

The corresponding author gratefully acknowledges KZ for Pesticides and Chemicals Company, Nubaria City, Egypt, for its valuable support.

Bazı bitki koruma ürünlerinde etkin madde hekzaflumuron'un tespiti için doğru ve güvenilir HPLC-UV tabanlı yöntem

Abstract

Bu çalışmanın temel amacı, özellikle meyve ve sebzeler olmak üzere bitkileri korumak için yaygın olarak kullanılan bir insektisit olan hekzaflumuron (HFM) konsantrasyonlarını ölçmek için yeni bir seçici, hassas ve doğrulanmış RP-HPLC analitik yöntemi sunmaktır. Miktar tayini, bir UV dedektörü ile donatılmış ters fazlı bir HPLC sistemi kullanılarak gerçekleştirildi. Yeni yöntemin geliştirilmesi, sabit sıcaklık 30 ºC'de ters fazlı C-18 (paslanmaz çelik, 5 µm, 250 × 4,6 mm) kolonda gerçekleştirildi. Hareketli faz, hacimsel oran 85:15, akış hızı 1 mL/dk ve algılama dalga boyu 220 nm'de asetonitril ve damıtılmış sudan oluşur. Ayırmanın tutulma süresi 3,84 dakikadır. Yöntem, CIPAC'a (Collaborative International Pesticides Analytical Council) göre özgüllük, doğrusallık, kesinlik, geri kazanım, LOD, LOQ ve doğruluk test edilerek doğrulandı ve SANCO/3030/99 rev.5 yönergelerinin yönergelerine uygundu. Yöntem, kabul edilebilir bir doğrusallık regresyonu R2 (0,9974) ortaya koydu. Yöntemin 50-250 µg/mL konsantrasyon seviyelerinden itibaren yüksek doğrulukla (%99,3-100,7) doğru olduğu bulundu. Yöntemin doğrulama sonuçları, bu aktif bileşeni içeren Emülsiyon Konsantre formülasyonlarının değerlendirilmesinde bir standardizasyon aracı olarak kullanılmasını uygun hale getirmektedir.

Keywords

Hexaflumuron , HPLC Analizi , Yöntem doğrulama , Hassasiyet , Doğruluk

References

• C. Lu, X. Yin, X. Liu, M. Wang, Study of the photodegradation kinetics and pathways of hexaflumuron in liquid media, Photochem. Photobiol., 90, 2014, 1219-1223.
• A. A. Mohamed, G. S. Refae, Effect of Hexaflumuron on Some Biological Aspects of The Predator Mite Eusebius Hutu (Pritchard & Baker), Egypt J. Agric Res, 91, 2013, 111-118.
• C. Tomlin, The pesticide manual: a world compendium, incorporating the agrochemicals handbook (10th edition), 1994, UK, British Crop Protection Council Publications.
• M. Mahmoudvand, H. Abbasipour, A. S. Garjan, A. R. Bandani, Sublethal effects of hexaflumuron on development and reproduction of the diamondback moth, Plutella xylostella (Lepidoptera: Yponomeutidae), Insect Sci, 18, 2011, 689-696.
• C. Yu, M. Fu, R. Lin, Y. Zhang, L. Yongquan, H. Jiang, T. C. Brock, Toxic effects of hexaflumuron on the development of Coccinella septempunctata, Environ Sci Pollut Res, 21, 2014, 1418-1424.
• S. Khajepour, H. Izadi, M. J. Asari, N. Liu, Evaluation of two formulated chitin synthesis inhibitors, hexaflumuron and lufenuron against the raisin moth, Ephestia figulilella, J insect sci, 12, 2012, 102.
• M. F. Al-Hakkani, N. Ahmed, M. H. Hassan, Rapidly, sensitive quantitative assessment of thiopental via forced stability indicating validated RP-HPLC method and its in-use stability activities, Sci Rep, 13, 2023, 10294.
• O. Coskun, Separation techniques: chromatography, North Clin Istanb, 3, 2016, 156-160.
• J. H. Kim, J. S. Seo, J. K. Moon, J. H. Kim, Multi-residue method development of 8 benzoylurea insecticides in mandarin and apple using high performance liquid chromatography and liquid chromatography-tandem mass spectrometry, J Korean Soc Appl. Biol Chem, 56, 2013, 47-54.
• L. Chen, J. Chen, Y. Guo, J. Li, Y. Yang, L. Xu, F. Fu, Study on the simultaneous determination of seven benzoylurea pesticides in Oolong tea and their leaching characteristics during infusing process by HPLC–MS/MS, Food Chem, 143, 2014, 405–410.
• C.H. Wang, X.X. Ma, C. Wang, Q.H. Wu, Z. Wang, Poly (vinylidene fluoride) membrane based thin film microextraction for enrichment of benzoylurea insecticides from water samples followed by their determination with HPLC, Chin Chem Lett, 25, 2014, 1625-1629.
• CIPAC (Collaborative International Pesticide Analytical Council), Guidelines on method validation to be performed in support of analytical methods for agrochemical formulations, Document no 3807, July 2003.
• European Commission (Directorate General Health and Consumer Protection), Technical Active Substance and Plant protection products: guidance for generating and reporting methods of analysis in support of pre-and post-registration data requirements for Annex (Section 4) of Regulation (EU) No 283/2013 and Annex (Section 5) of Regulation (EU) No 284/2013, SANCO/3030/99 rev.5, March 2019.
• M. Yuwono, G. Indrayanto, Validation of chromatographic methods of analysis, Profiles Drug Subst Excip Relat Methodol, 32, 2005, 243-259.
• I.M. Elkamhawy, A new Validated Method for Rapid Determination of Commercial Formulations Containing Quizalofop-P-Ethyl by Reversed-Phase Liquid Chromatography, SciRad, 3, 2024, 254-265.
• A. Sharma, J. K. Dubey, S. Katna, D. Shandil, G. S. Brar, S. Singh, Validation of analytical methods used for pesticide residue detection in fruits and vegetables, Food Anal Methods, 14, 2021, 1919-1926.
• B.T. Alquadeib, Development and validation of a new HPLC analytical method for the determination of diclofenac in tablets, Saudi Pharm J, 27, 2019, 66-70.
• L. Velkoska-Markovska, J. Davitkova, M. S. Jankulovska, HPLC Method For Determination of Active Ingredients in Pesticide Formulation SWITCH 62,5 WG, J Agric Food Environ Sci, 75, 2021, 11-18.
• B.M. Marson, V. Concentino, A.M. Junkert, M.M. Fachi, R.O. Vilhena, R. Pontarolo, Validation of analytical methods in a pharmaceutical quality system: an overview focused on HPLC methods, Quim Nova, 43, 2020, 1190-1203.
• I.M. Elkamhawy, Development and validation of an innovative HPLC-UV method for determination of Metalaxyl-M in flowable suspension fungicide formulas, Curr Chem Lett, in press, 2025.