Applied Chemical Engineering

Optimization of dispersive liquid–liquid microextraction (DLLME) and spectrophotometric analysis of sodium diclofenac in pharmaceutical formulations using Box-Behnken design (BBD)

Nibras Sabah Mohammed Ahmed

Chemistry Department, College of Science, Mustansiriyah University, Baghdad, 10045, Iraq

Muhammad Zaboon Thani

Chemistry Department, College of Science, Mustansiriyah University, Baghdad, 10045, Iraq

DOI: https://doi.org/10.59429/ace.v8i3.5658

Keywords: microextraction; sodium diclofenac; green method; box-behnken design; dllme method

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Abstract

A green and efficient Dispersive Liquid-Liquid Microextraction (DLLME) method coupled with spectrophotometry was developed and validated for the determination of sodium diclofenac in pharmaceutical samples. The method is based on the formation of an ion-pair complex to facilitate extraction. Critical extraction parameters, including pH, type and volume of extraction/disperser solvents, and centrifugation parameters, were systematically optimized using a Box-Behnken Design (BBD). This BBD approach enabled a comprehensive evaluation of the influence of these variables on extraction efficiency. Under optimal conditions, the method demonstrated excellent linearity within the range of 1.0-16.0 mg/L (R² = 0.9982). The limits of detection (LOD) and quantification (LOQ) were determined to be 0.326 mg/L and 0.987 mg/L, respectively, indicating good sensitivity. Comparative validation against a standard HPLC method, assessed using F-test and T-test, showed no statistically significant difference, thereby confirming the proposed method's accuracy and reliability. Overall, the proposed DLLME method offers a simple, efficient, environmentally friendly, and cost-effective approach for the accurate quantitative analysis of sodium diclofenac in pharmaceutical formulations.

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References

[1]. Dupra, K. J. B., & MA, R. S. A. Bioactivities of Purple Shamrock (Oxalis Triangularis) Crude Extract and Evaluation of Shamrock Topical Antibacterial Gel. (2024).

[2]. Ayub, R., Dar, A., & Anwar, J. Determination of Diclofenac Sodium in Pharmaceutical Preparations by Computational Image Scanning Densitometry. Journal of The Chemical Society of Pakistan, 45(6), 237. (2023).

[3]. Satar, H. A., Yousif, E., & Ahmed, A. DICLOFENAC: A Review on Its Synthesis, Mechanism of Action, Pharmacokinetics, Prospect and Environmental Impact. Al-Mustaqbal Journal of Pharmaceutical and Medical Sciences, 2(3), 2. (2024).

[4]. Demirhan, Y., & Koşal, V. The Effect of Diclofenac Sodium on Oxidative Stress and Some Reproductive Hormones in Rat Uterus. Fırat Üniversitesi Sağlık Bilimleri Veteriner Dergisi, 38(2). (2024).

[5]. Sami, I., Monir, H. H., Mahmoud, A. M., El-Saharty, Y. S., & Rostom, Y. Solid-state microfabricated potentiometric sensors for selective assessment of diclofenac sodium utilizing pentafluorophenyl diazonium modified carbon nanotubes transducer layer. Journal of The Electrochemical Society, 171(7), 077505. (2024).

[6]. Tuyet, N. T. A., Trinh, N. N., Bao, T. N., & Luan, T. A. K. Determination of activation energy of decomposition reaction from thermo gravimetric analysis. International Journal of Advances in Engineering and Management, 6(7), 486-491. (2024).

[7]. Mahmood, R. M., Darweesh, S. A., Alassaf, N. A., & Al-Khalisy, R. S. Simultaneous Spectrophotometric Determination of Piroxicam, Naproxen, Diclofenac Sodium and Mefenamic Acid in Pharmaceutical Formulations by Partial Least Squares Method. Methods & Objects of Chemical Analysis/Metody & Obekty Himičeskogo Analiza, 19(2). (2024).

[8]. Elkady, E., Fouad, M., & Mozayad, A. Application of Box–Behnken Design and Response Surface Methodology for Selecting the Optimum RP-HPLC Conditions for the Simultaneous Determination of Paracetamol and Diclofenac Sodium Along With Three Skeletal Muscle Relaxants in Three Different Pharmaceutical Dosage Forms. Journal of Chromatographic Science, 62(4), 328-338. (2024).

[9]. AlNahwa, L. H., Ali, H. M., Hasanin, T. H., Shalaby, K., Alshammari, M. S., Alsirhani, A. M., & Mohamed, S. H. Development of an RP-HPLC Method for Quantifying Diclofenac Diethylamine, Methyl Salicylate, and Capsaicin in Pharmaceutical Formulation and Skin Samples. Molecules, 29(12), 2732. (2024).

[10]. Patel, K., Verriboina, S. K., & Vasantharaju, S. G. Stability indicating assay method development and validation of simultaneous estimation of chlorzoxazone, diclofenac sodium and paracetamol in bulk drug and tablet by RP-HPLC. Research Journal of Pharmacy and Technology, 14(9), 5024-5028. (2021).

[11]. Şarkaya, K., Fitoz, A., Ergun, E., Orhan, E., & Ergun, Ü. A Fluorometric Method for Diclofenac Determination in Liquid Formulations: Experimental and Computational Assessments. ChemistrySelect, 10(1), e202404083. (2025).

[12]. Bakhoum, J. M., Mbaye, O., Bakhoum, J. P., Gaye-Seye, M. D., Trellu, C., Atanasse, C., & Aaron, J. J. Diclofenac analysis in natural waters using UV-visible absorption spectrometric method. Macedonian Journal of Chemistry and Chemical Engineering, 42(2), 145-152. (2023).

[13]. Mukkawi, R., Shantier, S., & Gadkariem, E. A. Spectrophotometric method for the simultaneous analysis of diclofenac sodium and lidocaine hydrochloride in bulk and dosage forms. Pharmaceutical Chemistry Journal, 55, 831-834. (2021).

[14]. Ghosal, P. In Vitro Evaluation of Some Brands of Diclofenac Sodium Injection Marketed in Greater Noida, Uttar Pradesh, India Using UV-Vis Spectrophotometric Method. Current Research in Pharmaceutical Sciences, 178-185. (2022).

[15]. Kareem, N. K., Thani, M. Z., & Al-Rawi, K. F. New approach for determination of Phenylephrine HCl in Pure and Pharmaceutical Formulation using a various Microextraction Methods. Research Journal of Pharmacy and Technology, 15(4), 1648-1652. (2022).

[16]. Nadi, Z., Rahbar-Kelishami, A., & Shayesteh, H. An effective dispersive liquid-liquid microextraction method for pharmaceutical extraction: Optimization via central composite design. Scientia Iranica, 31(9), 693-703. (2024).

[17]. Salim, A. S., Thani, M. Z., Atia, A. J. K., & Fattah, H. A. A Simple Dispersive Liquid-Liquid Microextraction for the Determination of Copper II in the Water Samples of the Halfaya Oil Field in Maysan. Al-Mustansiriyah Journal of Science, 35(3), 58-73. (2024).

[18]. Jabbar, A. I., Thani, M. Z., & Khaleel, A. I. Ion-Pair Complex of Trimethoprim with Alizarin Yellow and its Determination and Extraction by LLE and DLLME. Tikrit Journal of Pure Science, 28(3). (2023).

[19]. Jabbar, A. I., Khaleel, A. I., & Thani, M. Z. Determination of Cefixime inclusion complex with 2-hydroxypropyl-β-cyclodextrin and extraction using DLLME and SIHLLME. Journal of University of Anbar for Pure Science, 17(1). (2023).

[20]. Ghane, M., Ezoddin, M., Mohadesi, A., & Karimi, M. A. Reversible switchable deep eutectic solvent for dispersive liquid–liquid microextraction of non-steroidal anti-inflammatory drugs in biological and wastewater samples combined with high-performance liquid chromatography. Microchemical Journal, 203, 110915. (2024).

[21]. Hammad, S. F., Abdallah, I. A., Bedair, A., & Mansour, F. R. Homogeneous liquid–liquid extraction as an alternative sample preparation technique for biomedical analysis. Journal of separation science, 45(1), 185-209. (2022).

[22]. Thani, M. Z., Dadoosh, S. A., Abdullah, A. M., Fahad, A. S., Fahad, Y. S., & Faraj, F. L. Evaluation of salbutamol in pure form and pharmaceutical formulations using spectrophotometry and green nonionic surfactant of cloud point extraction. In Journal of Physics: Conference Series, 1853(1), 012022. (2021).

[23]. Dadoosh, S. A., Thani, M. Z., Abdullah, A. M., Fahad, A. S., & Fahad, Y. S. Development of an ecological-friendly method for dexamethasone determination and cloud point extraction in pharmaceutical formulations using schiff base reaction. Egyptian Journal of Chemistry, 64(9), 5083-5092. (2021).

[24]. Al_Saadi, M. R., Al-Garawi, Z. S., & Thani, M. Z. Promising technique, cloud point extraction: Technology & applications. In Journal of Physics: Conference Series, 1853(1), 012064. (2021).

[25]. Samadi-Maybodi, A., & Rahmati, A. Dual metal zeolitic imidazolate frameworks as an organometallic polymer for effective adsorption of chlorpyrifos in aqueous solution. Environmental 1 Engineering Research, 25(6

[26]. Tamandani, M., & Hashemi, S. H. Central composite design (CCD) and Box-Behnken design (BBD) for the optimization of a molecularly imprinted polymer (MIP) based pipette tip micro-solid phase extraction (SPE) for the spectrophotometric determination of chlorpyrifos in food and juice. Analytical Letters., 55 (15), 2394-2408 (2022).

[27]. Demir, A., Fesliyan, S., Altunay, N., & Soylak, M. Deep eutectic solvent-based sonication assisted dispersive liquid-liquid microextraction using Box-Behnken optimization for the determination of patent blue V in food and drug samples. Journal of Food Composition and Analysis., 135, 106634 (2024).

[28]. Kadhim, S. S., Abbas, R. F., & Jaafar, S. S. Removal of Cr (VI) from aqueous solution by using polyaniline/polycarbonates nanofibers composite: central composite design, isotherm, and error analysis. Desalination and Water Treatment., 229, 343-351 (2021).

[29]. Singh, R., Singh, P., Pandey, V. K., Dash, K. K., Ashish, Mukarram, S. A., ... & Kovács, B. Microwave-assisted phytochemical extraction from walnut hull and process optimization using Box–Behnken design (BBD). Processes., 11 (4), 1243 (2023).

[30]. Isadiartuti, D., Rosita, N., Hendradi, E., Putri, F. F. D. P., & Magdalena, F. Solubility and partition coefficient of salicylamide in various ph buffer solutions. Indonesian Journal of Chemistry., 21 (5), 1263-1270 (2021).

[31]. Emara, L. H., Taha, N. F., El-Ashmawy, A. A., Raslan, H. M., & Mursi, N. M. (2012). A rapid and sensitive bioanalytical hplc method for determining diclofenac sodium in human plasma for bioequivalence studies. Journal of liquid chromatography & related technologies, 35(15), 2203-2216.

[32]. da Silva Lyra, W., Sanches, F. A. C., da Silva Cunha, F. A., Diniz, P. H. G. D., Lemos, S. G., da Silva, E. C., & de Araujo, M. C. U. (2011). Indirect determination of sodium diclofenac, sodium dipyrone and calcium gluconate in injection drugs using digital image-based (webcam) flame emission spectrometric method. Analytical Methods, 3(9), 1975-1980.

[33]. Chohan, M. S., Elgorashe, R. E. E., Balgoname, A. A., Attimarad, M., SreeHarsha, N., Venugopala, K. N., ... & Pottathil, S. (2019). Eco-friendly Derivative UV Spectrophotometric Methods for Simultaneous Determination of Diclofenac Sodium and Moxifloxacin in Laboratory Mixed Ophthalmic Preparation. Indian J. Pharm. Educ. Res, 54, 166-174.

[34]. Bhatt, N. M., Chavada, V. D., Sanyal, M., & Shrivastav, P. S. (2014). Manipulating ratio spectra for the spectrophotometric analysis of diclofenac sodium and pantoprazole sodium in laboratory mixtures and tablet formulation. The scientific world journal, 2014(1), 495739.

[35]. Yilmaz, B., & Ciltas, U. (2015). Determination of diclofenac in pharmaceutical preparations by voltammetry and gas chromatography methods. Journal of Pharmaceutical Analysis, 5(3), 153-160.

[36]. Jalbani, N. S., Solangi, A. R., Khuhawar, M. Y., Memon, S., Junejo, R., & Memon, A. A. (2020). Gas chromatographic and spectrophotometric determination of diclofenac sodium, ibuprofen, and mefenamic acid in urine and blood samples. Turkish Journal of Pharmaceutical Sciences, 17(5), 465.

[37]. Madikizela, L. M., & Chimuka, L. (2017). Simultaneous determination of naproxen, ibuprofen and diclofenac in wastewater using solid-phase extraction with high performance liquid chromatography. Water sa, 43(2), 264-274.