HPTLC methods to assay active ingredients in pharmaceutical formulations: A review of the method development and validation steps (original) (raw)
Related papers
Calibration and Validation of Analytical Methods - A Sampling of Current Approaches, 2018
Method validation is an important activity for pharmaceutical evaluations to ensure that analytical methods are suitable for their intended use. With particular focus on active ingredient and impurities, the implementation of different categories of method validation are explained for qualitative and quantitative methods. Detailed explanations with example approaches are provided for the key aspects of method validation, namely specificity, accuracy, linearity, limits of detection/quantitation, precision, robustness, and method range. While all of the sections outlined for method validation are generally applicable for a variety of techniques commonly used in pharmaceutical analysis (i.e., UV and HPLC instrumentation), focused attention is provided for examples that have been implemented using high performance thin layer chromatographic techniques.
Analysis of pharmaceutical and natural compounds and newer drugs is commonly used in all the stages of drug discovery and development process. HPTLC is an advanced type of planar chromatography used extensively in the recent years for fingerprinting of medicinal plants, products and for screening lichen substances. Major advantages include precise sample application, wide choice of stationary phases, simultaneous processing of standards, simultaneous scanning in different light sources and excellent documentation facility. It is more sensitive and possible to run more samples in a short period of time, by using small amount of solvent. It is one of the sophisticated instrumental techniques based on the full capabilities of thin layer chromatography. In the present review brief discussion is presented with regard to type of instrument used in HPTLC, its entire methodology and how this technique is better than TLC. This concise information consists of salient points showing immense use in the practical analysis of HPTLC. This article consists of compilation of 63 formulations in total, out of which 38 containing one active substances and 25 containing dual active substances are shown in separate Tables (Table 1 and 2). The names of products, solvent system, details of plates to be used and details of quantitation showing densitometric evaluation are shown against each formulation.
Journal of Chromatography A, 2013
The accuracy profile, based on total error, integrates several validation parameters, such as trueness, precision and linearity, providing one statistic which enables decision on the suitability of a method for its intended purpose. Two assay methods for formulations are validated using accuracy profiles as an alternative approach to classic method validation. It concerns high-performance thin-layer chromatography (HPTLC) methods, which initially were validated using the classic approach. The first method assayed sulfamethoxazole and trimethoprim, and the second lamivudine, stavudine and nevirapine. Both formulations are fixed-dose combination tablets. The resulting accuracy profiles showed that the 95% expectation tolerance limits for all compounds fell well within the bias acceptance limits set at ±5%. This means that the two analytical thin-layer chromatographic methods are capable of making accurate results at the studied concentration ranges of each compound. Measurement uncertainties of every compound at each concentration level could also be determined from the accuracy profile data.
American Journal of Analytical Chemistry, 2023
A simple and rapid HPTLC analytical method has been developed and validated for the determination of Etanercept and Filgrastim in pure form and in marketed formulation. Both the drugs were chromatographed on silica gel 60 F254s HPTLC plates, as stationary phase. The mobile phase optimized for Filgrastim and Etanercept was Water: n-butanol (7.5:2.5 v/v) and Isopropyl alcohol: water (6.5:4.5 v/v), respectively. The chromatogram obtained was scanned at 225 nm and 222 nm for filgrastim and etanercept which resulted in a retention factor of 0.45 ± 0.07 and 0.32 ± 0.03, respectively. The method was validated for parameters like linearity, accuracy, precision, specificity and robustness. Recovery studies were performed at three concentration levels, to demonstrate suitability, accuracy and precision of proposed method. Statistical analysis proved that the proposed method is accurate and reproducible with linearity in the range of 500 to 3000 ng/band for filgrastim and 200 to 1200 ng/band for etanercept. The limit of detection and limit of quantification for filgrastim was found to be 63.418 ng/band and 192.177 ng/band. For etanercept, LOD and LOQ were found to be 33.381 ng/band and 101.153 ng/band, respectively. The proposed method can be employed for the routine analysis of selected biosimilars.
Critical Reviews in Analytical Chemistry, 2012
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. The publisher does not give any warranty express or implied or make any representation that the contents will be complete or accurate or up to date. The accuracy of any instructions, formulae, and drug doses should be independently verified with primary sources. The publisher shall not be liable for any loss, actions, claims, proceedings, demand, or costs or damages whatsoever or howsoever caused arising directly or indirectly in connection with or arising out of the use of this material.
A Comprehensive Working, Principles and Applications of Thin Layer Chromatography
2014
In this present article, we address the basic aspects such as idea, mechanism and working of Thin layer Chromatography (TLC) in analytical as well as preparative preparation methods. We have gone through diverse journals for gathering complete package of TLC and found that TLC is very simple, easy, less time consuming, costeffective and multiple samples could be run in one go hence, is constantly the first choice for varied application in qualitative analysis of pharmaceutical products. In this modern scientific world were HPLC and HPTLC technology has developed still TLC holds good promise for identification and analysis of different bioactive compounds, secondary metabolites, Vitamins and amino acids. It is a very preliminary analytical method done prior to HPLC and reaction progress can be monitored easily. It can be used for separating compounds from crude extracts and separating impurities from a compound. Identical compounds from the mixture can be easily separated by analytic...
2015
Many different strategies of high performance liquid chromatographic method development are being used today. This overview describes a strategy for the systematic development of High performance liquid chromatographic (HPLC) methods. It is an analytical tool which is able to separate, detect and quantify the drug, its various impurities and drug related degradants that can form on synthesis or storage. HPLC involves the understanding of chemistry of drug substance and facilitates the development of the analytical method. Many chromatographic parameters were evaluated in order to optimize the method. Appropriate mobile phase, stationary phase, column, column size, temperature, wavelength and gradient must be found that affords suitable compatibility and stability of drug as well as impurities and degradants. In this paper we have discussed the different physical and chemical parameters that govern the HPLC process and operation and suggested method development for the most optimum c...
Recent applications of analytical techniques for quantitative pharmaceutical analysis: A review
2010
The intension of this paper was to review and discuss some of the current quantitative analytical procedures which are used for quality control of pharmaceutical products. The selected papers were organized according to the analytical technique employed. Several techniques like ultraviolet/visible spectrophotometry, fluorimetry, titrimetry, electroanalytical techniques, chromatographic methods (thin-layer chromatography, gas chromatography and high-performance liquid chromatography), capillary electrophoresis and vibrational spectroscopies are the main techniques that have been used for the quantitative analysis of pharmaceutical compounds. In conclusion, although simple techniques such as UV/VIS spectrophotometry and TLC are still extensively employed, HPLC is the most popular instrumental technique used for the analysis of pharmaceuticals. Besides, a review of recent works in the area of pharmaceutical analysis showed a trend in the application of techniques increasingly rapid such as ultra performance liquid chromatography and the use of sensitive and specific detectors as mass spectrometers.
High performance liquid chromatographic methods (HPLC)
2002
Figure 4-Chromatogram of freshly coated tablets before drying, showing both methanol and methylene chloride after 15 min of coating. Kpy: (2) methanol; (2) methylene chloride. Statistical limits of detection were established for methanol a t 9.5 f 1.5 ppm per average tablet weight and for methylene chloride a t 10.1 f 2.3 ppm per tablet. Below these limits, sensitivity of the instrument was unreliable. All absolute values were approximate due to two factors. First, precision of measurement is decreased as concentrations used in the calibration curve approach the detection limit. As an alternative, a calibration curve could have been drawn at higher concentrations and extrapolated to the limit of detection (3). Second, this detection limit is also dependent on where the confidence lines are drawn about the regression line and is, a t best, an estimate for a limit of guaranteed purity (2). Therefore, the values reported here represented a range within a specified precision for the standards, where the analytical method and the instrumentation were reliable for low-level determinations (6). The levels of solvent residues, ranging from 30 to 300 ppm, obtained during the film-coating process were statistically significant. However, after 24 h of drying, no peaks for either solvent were observed in the chromatograms. This procedure is easy and reliable for monitoring organic residues on film-coated tablets. Determining lower limits of detection should be an integral part of any statistical package developed for a new analytical method. REFERENCES
JURNAL FARMASI DAN ILMU KEFARMASIAN INDONESIA
Background: Chromatography is one of the analytical techniques widely used for the quality control process in the pharmaceutical industry. One of the analytical methods used in drug analysis is Thin Layer Chromatography (TLC). The analysis process of TLC can be performed using densitometry (scanner) or videodensitometry (videoscan). The principal analysis of densitometry (scanner) is based on the density measured from each spot on the TLC plate using a specific wavelength range, and videodensitometry (videoscan) is performed by taking pictures of the plate using a Visualizer at a specific wavelength. Objective: This review article discusses the application of densitometry and videodensitometry methods for quantitative analysis of pharmaceutical products. Methods: This study was conducted using a systematic review method using the PRISMA statement from January to April 2023. Four databases were searched: PubMed, ScienceDirect, Scopus, and Google Scholar with inclusion criteria: studi...