Validation of a Thin-Layer Chromatography for the Determination of Hydrocortisone Acetate and Lidocaine in a Pharmaceutical Preparation

Abstract

A new specific, precise, accurate, and robust TLC-densitometry has been developed for the simultaneous determination of hydrocortisone acetate and lidocaine hydrochloride in combined pharmaceutical formulation. The chromatographic analysis was carried out using a mobile phase consisting of chloroform + acetone + ammonia (25%) in volume composition 8 : 2 : 0.1 and silica gel 60F254 plates. Densitometric detection was performed in UV at wavelengths 200 nm and 250 nm, respectively, for lidocaine hydrochloride and hydrocortisone acetate. The validation of the proposed method was performed in terms of specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, and robustness. The applied TLC procedure is linear in hydrocortisone acetate concentration range of 3.75 ÷ 12.50 μg·spot−1, and from 1.00 ÷ 2.50 μg·spot−1 for lidocaine hydrochloride. The developed method was found to be accurate (the value of the coefficient of variation CV [%] is less than 3%), precise (CV [%] is less than 2%), specific, and robust. LOQ of hydrocortisone acetate is 0.198 μg·spot−1 and LOD is 0.066 μg·spot−1. LOQ and LOD values for lidocaine hydrochloride are 0.270 and 0.090 μg·spot−1, respectively. The assay value of both bioactive substances is consistent with the limits recommended by Pharmacopoeia.

Development of a simple, low-cost and rapid thin-layer chromatography method for the determination of individual volatile fatty acids

“…Finally, relative standard deviation (RSD) between 4.6 and 6.7 % were obtained after application of the whole protocol on a 50 mg.L -1 standard (n=7, with one TLC plate for each repetition). Precision with our method is therefore lower than that usually obtained with TLC scanner and dedicated data analysis software [18,26], but is satisfactory enough for our analytical purpose using low-cost instruments with easy to find materials and software.…”

“…Among separation techniques, Thin-Layer Chromatography (TLC) is the oldest one and certainly the simplest and least expensive tool in analytical chemistry. Many studies have already proposed TLC as an alternative to more modern and complex chromatographic system for quality control and fast quantitative or semi-quantitative determination [18][19][20][21].…”

Nanogram/mL detection of nortriptyline: Preparation of a molecularly imprinted polymer for spectrophotometric determination of nortriptyline based on multivariate optimization methods

“…According to the calibration curve, the absorbance was increased linearly with the concentration increasing in the range of 0.1–100 µmol/L (0.026–26 µg/mL) of NOR, described by linear regression equation ( R 2 = 0.9953) as follows: where, C is NOR concentration (mol/L) and, A is the UV absorbance at 239 nm. The LOD was calculated 1.03 × 10 −8 mol/L (2.7 ng/mL) based on 3.3 sigma method (3.3σblank/sensitivity; n = 5) . In addition, the repeatability was evaluated for three times measurements of NOR standard solution with the concentration of 5 × 10 −5 mol/L (relative standard deviation of 3.7%).…”

“…where, C is NOR concentration (mol/L) and, A is the UV absorbance at 239 nm. The LOD was calculated 1.03 × 10 −8 mol/L (2.7 ng/mL) based on 3.3 sigma method (3.3σblank/sensitivity; n = 5) [28]. In addition, the repeatability was evaluated for three times measurements of NOR standard solution with the concentration of 5 × 10 −5 mol/L (relative standard deviation of 3.7%).…”

Extraction-Spectrophotometric Determination of Lidocaine Hydrochloride in Pharmaceuticals

“…Various analytical techniques for LD-HCL determination in biological and pharmaceutical samples have been published, including the chromatographic methods,gas chromatography(GC) determination of LD-HCl in whole blood (Keenaghan, 1968), (Edhorn, 1971), (Asada, 1979) (Hattori,Yamamoto, & Yamada, 1991) (Watanabe, Namera, Yashiki, Iwasaki, & Kojima, 1998), from serum (Kruczek, 1981) (Prat , & Bruguerolle, 1986) (Baniceru, Croitoru, & Popescu, 2004), from plsma (Björk , Pettersson, & Osterlöf, 1990), in human plasma and urine (Ohshima& Takayasu, 1999), in human urine (Koster, Hofman, & Jong, 1998), to distinguish the drug from its thermal degradation produts (Stavchansky, Eghbali & Geary, 1987), in pharmaceutical preparations (Tarli P, Benocci & Neri, 1969), and Forced Degradation of Lidocaine HCl by NMR Spectroscopy and GC-FID Methods (Kadioglu, Atila, Serdar, Gultekin, & Alcan, 2013), thin-layer chromatography (TLC)for the Determination of Hydrocortisone Acetate and Lidocaine in a Pharmaceutical Preparation (Dołowy, Kulpińska-Kucia, & Pyka, 2014), high-performance liquid chromatography (HPLC) and TLC (Živanović, Živanov-Stakić, & Radulović, 1998) (Abdelwahab, Nouruddin, Abdelkawy, Emam, 2016), HPLC, in plasma with solid phase extraction and UV detection (Kang, Jun, & McCall, 1999 ), in pharmaceuticals (Smith & Nuessle, 1981) (Waraszkiewicz, Milano, & DiRubio, 1981) (Atay & Öztop, 1997) (Parissi-Poulou & Panderi, 1999) (Gebauer, McClure, & Vlahakis, 2001) (Liawruangrath, Liawruangrath, & Pibool, 2001) (Malenovic, Medenica, Ivanovic, Jancic, & Markovic, 2005) (Zivanovic, Zecevic, Markovic, Petrovic, & Ivanovic, 2005) (Salas, Talero, Rabasco, & González, 2008) (Stojanović, Malenović, Marković, Ivanović, & Medenica, 2010) …”