Molekul, Vol. 17. No. 2, July 2022: 252 – 260 252 MOLEKUL eISSN: 2503-0310 Articles https://doi.org/10.20884/1.jm.2022.17.2.6348 Chiral Separation of Econazole by High Performance Liquid Chromatography Method using Cyclodextrin as Chiral Column Dadan Hermawan 1 *, Cacu 1 , Khansa Salsabila 1 , Suwandri 1 , Amin Fatoni 1 , Uyi Sulaeman 1 , Ponco Iswanto 1 , Mudasir Mudasir 2 , and Hassan Y. Aboul-Enein 3 1 Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Jenderal Soedirman, Purwokerto, 53123, Indonesia 2 Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia 3 Department of Pharmaceutical and Medicinal Chemistry, Pharmaceutical and Drug Industries Research Division, National Research Center (NRC) Dokki, Cairo, 12622, Egypt *Corresponding author email: dadan.hermawan@unsoed.ac.id Received September 12, 2021; Accepted June 09, 2022; Available online July 20, 2022 ABSTRACT. The chiral separation of econazole, an antifungal drug with one chiral center has been successfully carried out using the high-performance liquid chromatography (HPLC) method. Enantioresolution of econazole (Rs = 2.29) was achieved using cyclodextrin-based chiral column (Astec Cyclobond, 25 cm × 4.6 mm × 5 μm), mobile phase composition of acetonitrile : water (0.2% HCOOH) (20:80, v/v), and UV detection of 220 nm. The optimized HPLC method has been applied for the quantitative determination of econazole in the pharmaceutical (liquid) sample with percentage recovery of 100.75 % (RSD = 0,95%; n = 3). The effect of several HPLC parameters on the chiral separation of econazole was also evaluated and the method was successfully validated in terms of linearity, accuracy, precision, and selectivity. The present HPLC method was simple, short analysis time, and high resolution. Keywords: Chiral separation, econazole, high-performance liquid chromatography. INTRODUCTION The enantiomeric separation is one of the most important fields in modern analytical chemistry, mainly in pharmaceutical and agrochemical products since a stereochemistry has a crucial role on biological activity. Enantiomers have the same chemical properties except to the reactivity of the enantiomers to optically active reagents. The physical properties of enantiomers are identical, but their direction e.g. rotation is different (Yardimci, 2020). Many biological and pharmacological compounds are asymmetrical showing optical activity (Sahu et al., 2018; Zhang et al., 2020). Administration of pure, pharmacologically, and pharmacokinetically active enantiomers are important. Enantioselective synthesis is the most ideal method for obtaining pure enantiomers. However, this method is not recommended because it is impractical and expensive. On the other hand, the demand for enantiomeric separation methods on an analytical scale to control synthesis, check the racemization process, control the enantiomeric purity, and pharmacokinetic studies are very urgent at this time (Zhang et al., 2020; Dubey et al., 2012). The physical and chemical properties of the enantiomers are identical in the symmetrical environment but differ in the pharmacological properties of the receptor. Enantiomeric separation is carried out to determine one of the enantiomers of a stereoisomer that has active drug properties. Most of the synthetic drugs still contain chiral compounds that are still marketed in racemic conditions and only a small portion are pure enantiomers. This racemic condition means that a drug in addition to having active enantiomeric properties also has enantiomeric properties that are inactive with some unwanted side effects or that it is toxic (Valimana-Traverso et al., 2019). The antifungal drug imidazole has two nitrogen atoms in the azole ring. Butoconazole, ketoconazole, bifonazole, isoconazole, clotrimazole, oxiconazole, fenticonazole, miconazole, and econazole are some of the examples of this azole class (Yardimci, 2020). Econazole has a chemical name 1-[2- (2,4-dichlorophenyl) -2-(4-chlorobenziloxy) -ethyl]- imidazole with the molecular formula C18H15C13N20 (Salido-Fortuna et al., 2020). Currently, imidazole antifungals are widely used in the treatment of