Selective High Performance Thin Layer Research Articles

HPTLC determination of ergosterol in wheat and structure elucidation by NMR: Toward confirming method selectivity

Ergosterol (ERG) is considered as an excellent fungal biomarker, therefore it was utilized for the determination of the degree of fungi infection in wheat grains. A novel, fast, cost-effective and selective high performance thin layer chromatography (HPTLC) with reflectance/absorbance densitometry method was developed and validated for ERG determination. Separation was attained on pre-coated silica gel HPTLC plate with ethyl acetate: petroleum ether (6: 4 v/v) as a mobile phase. This system gave compact bands at retardation factor (Rf) of 0.45 ± 0.002 and quantification was performed at 282 nm over concentration range of 40–600 ng/band. The lower detectable limit was 10.79 ng per band which is three times more sensitive than the previously reported TLC method. The developed method could be utilized efficiently for the detection of ERG in pure powders and in fungi infected wheat samples. Resolution of ERG from other compounds in wheat grain extract allowed utilizing preparative TLC for separating and purifying ERG in a sufficient quantity for structure elucidation by nuclear magnetic resonance spectroscopy (NMR) for the first time. H1 and C13 NMR spectra indicated the purity of ERG extracted from the scrapped silica of preparative TLC plate at the Rf of the standard ERG verifying the method selectivity.

HPTLC method development and validation for the determination of andrographolide in raw material and tablet containing ethyl acetate fraction of Andrographis paniculata

Purpose: To develop and validate a rapid, accurate, and selective High Performance Thin Layer Chromatography (HPTLC), for quantification of andrographolide in raw materials and tablets ethyl acetate fraction of Andrographis paniculata as antimalarial agent.
 Methods: HPLC was conducted to determine the andrographolide concentration of ethanol extract and Camag linomat 5 in the stationary phase. HPLC measurements were conducted at a wavelength of 228 nm with a ratio of chloroform: methanol (90:10, v/v) in the mobile phase.
 Results: The validated method was separated andrographolide from other component with good resolution, and obtained retention factor was 0.38 ± 0.03. The data for calibration plot showed good linear relationship, with R2 = 0.998 in the concentration range of 138.0 - 460.0 ng/spot. The limit of detection and quantification were 9.6 ng/spot and 28.8 ng/spot, respectively. The percentage recovery was between 98.0 and 100.5 %. Additionally, the relative standard deviation method was between 1.4 and 1.0 %
 Conclusion: This method fulfills the validation requirements of selectivity, linearity, accuracy, and precision. Further, it can separate andrographolide from degradants. Thus, HPTLC method can be used to analyze the ethyl acetate fraction of ethanol extract of A. paniculata and its tablet products.

HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHIC DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF ZIPRASIDONE HYDROCHLORIDE MONOHYDRATE FROM BULK AND IT'S ORAL FORMULATION

A simple and selective high performance thin layer chromatographic (HPTLC) method was developed and validated for the estimation of ziprasidone hydrochloride monohydrate (ZHM) from its marketed formulation and oral formulation. The CAMAG HPTLC system, operated with software winCATS (ver.1.4.1.8) was used for proposed analytical work. Planar chromatographic development was carried out with the help of silica gel 60 F254 precoated aluminium TLC plates. Sample application was employed using Linomat 5 applicator. The optimized mobile phase was composed of toluene: glacial acetic acid: methanol (6: 0.3: 3 V/V/V). The detection of spots was carried out densitometrically using a UV detector at 317nm in absorbance mode. In HPTLC densitogram well defined peak was obtained for ZHM with starting position at 59 hRf, max position at 63 hRf and end position at 65 hRf. The optimum hRf value for ZHM was found to be 63. Performance characteristics of HPTLC method for estimation of ZHM in bulk drug and its marketed dosage form were statistically validated as per recommendations of ICH guidelines of analytical method validation. The HPTLC method was found to be linear across the range 20-140 ng/band. The LOD and LOQ values were found to be 13.573 and 41.130 ng/band, respectively. The method was found to be accurate, precise, robust and economical for the analysis of ZHM from bulk drug and its formulation. The proposed analytical HPTLC method can be used for routine quality-control analysis of ZHM.

DEVELOPMENT AND VALIDATION OF HPTLC METHOD FOR SIMULTANEOUS ESTIMATION OF TERBINAFINE HYDROCHLORIDE AND MOMETASONE FUROATE IN COMBINED DOSAGE FORM

A new simple, precise, accurate, specific and selective high performance thin layer chromatographic (HPTLC) method has been developed for the simultaneous estimation of Terbinafine hydrochloride (TH) and Mometasone furoate (MF) in cream dosage form. The chromatographic separation was achieved on Merck precoated silica gel aluminium plate 60 F254 using Toluene: Ethyl acetate: Glacial acetic acid (8: 4: 0.1 v/v) as mobile phase. The densitometric scanning was carried out at 248 nm. Response was found to be linear in the concentration range of 1000–3000 ng/band with correlation coefficient (r 2 = 0.999) for Terbinafine hydrochloride and 100-300 ng/band with correlation coefficient (r 2 = 0.998) for Mometasone furoate. All parameters of method, repeatability, precision, LOD, LOQ, % recovery were validated as per ICH guidelines. The proposed procedure was successfully applied for the simultaneous determination of both drugs in commercial cream preparation. The advantage of the method is simplicity, reasonable sensitivity, rapidity, excellent resolving power, low cost and is a more effective option than other chromatographic techniques in routine quality control.

Simultaneous determination of carbinoxamine maleate and pseudoephedrine HCl in their pure form and in their pharmaceutical formulation by HPTLC-densitometric method

An accurate, sensitive and selective High Performance Thin Layer Chromatography (HPTLC) method has been developed and validated for determination of carbinoxamine maleate and pseudoephedrine HCl in their binary mixture without previous separation. Chromatographic separation was performed on aluminum plates precoated with silica gel 60F 254 using acetone: methanol:ethylacetate:triethylamine (5:4:2:0.2, v : v : v : v ) as a developing system. The bands were scanned at 215 nm. The proposed method has been validated as per ICH guidelines and its linearity was evident in the ranges of 0.4-8.0 and 4-24 µg/ band for carbinoxamine maleate and pseudoephedrine HCl, respectively. The developed method has been applied for determination of the above mentioned drugs in their pharmaceutical formulation where no interference from methyl paraben present as excipient has been detected. Statistical comparison of the results obtained by the developed HPTLC method and those obtained by the reported HPLC method showed no significant difference between them. The developed method is sensitive, accurate and precise and can be easily used for quality control analysis of the studied drugs.

HPTLC Method for the Determination of Paracetamol, Pseudoephedrine and Loratidine in Tablets and Human Plasma.

A sensitive, accurate and selective high performance thin layer chromatography (HPTLC) method was developed and validated for the simultaneous determination of paracetamol (PAR), its toxic impurity 4-aminophenol (4-AP), pseudoephedrine HCl (PSH) and loratidine (LOR). The proposed chromatographic method has been developed using HPTLC aluminum plates precoated with silica gel 60 F254 using acetone-hexane-ammonia (4:5:0.1, by volume) as a developing system followed by densitometric measurement at 254 nm for PAR, 4-AP and LOR, while PSH was scanned at 208 nm. System suitability testing parameters were calculated to ascertain the quality performance of the developed chromatographic method. The method was validated with respect to USP guidelines regarding accuracy, precision and specificity. The method was successfully applied for the determination of PAR, PSH and LOR in ATSHI(®) tablets. The three drugs were also determined in plasma by applying the proposed method in the ranges of 0.5-6 µg/band, 1.6-12 µg/band and 0.4-2 µg/band for PAR, PSH and LOR, respectively. The results obtained by the proposed method were compared with those obtained by a reported HPLC method, and there was no significance difference between both methods regarding accuracy and precision.

Validated HPTLC method for the simultaneous determination of cinnarizine and dimenhydrinate in their combined dosage form

A simple, rapid and selective high performance thin layer chromatography (HPTLC) method was developed for the simultaneous determination of cinnarizine (CNZ) and dimenhydrinate (DMH) in pure form and in their combined dosage form. Reviewing the literature revealed that there are no reports for the use of TLC for the assay of this mixture. Effective separation was achieved using Fluka HPTLC aluminum sheets of silica gel 60 F254 using chloroform–n-hexane–methanol (8.5:0.8:0.7, by volume) as mobile phase, followed by densitometric measurement of CNZ and DMH spots at 254 and 279nm, respectively. The reliability and analytical performance of the proposed HPTLC method were statistically validated with respect to linearity, ranges, precision, accuracy, selectivity, robustness and detection and quantification limits. Calibration curves were linear in the ranges of 50–400 and 50–500ng/spot for CNZ and DMH, respectively with correlation coefficients >0.9998. The limits of detection were 8.1 and 8.0ng/spot for CNZ and DMH respectively. The validated HPTLC method was applied to the simultaneous analysis of CNZ and DMH in laboratory-prepared tablets. Both analytes were successfully quantified with good recovery values, and no interference was encountered from the inactive ingredients.

Development and validation of a high performance thin layer chromatographic method for determination of 1, 8-Cineole in Callistemon Citrinus.

A new, simple, precise, rapid, and selective high performance thin layer chromatographic (HPTLC) method has been developed and validated for the estimation of 1, 8-cineole in volatile oil of leaves of Callistemon Citrinus obtained by hydro distillation. The method was validated as per ICH guidelines and can be utilized for routine analysis. The retention factor for 1, 8-cineole was found to be 0.52. The linearity was found to be in the range of 3 μg-12 μg. The recovery obtained for 1, 8-cineole was 98%, which is satisfactory. The result obtained in validation indicate the accuracy, reproducibility, and reliability of the developed HPTLC method for determination of 1, 8-cineole.

Validated HPTLC method for simultaneous determination of loratadine and desloratadine in presence of Co-formulated drug

A highly validated and selective high performance thin layer chromatography (HPTLC) method was developed for the determination of loratadine (LOR) and desloratadine (DES) in their pharmaceutical formulations. The proposed method was based on HPTLC separation of the two drugs followed by densitometric measurements of their spots at 254 nm. The separation was carried out on Merck HPTLC aluminum sheets of silica gel 60 F254 using methanol-ammonia (10:0.3, ν/ν) as mobile phase. The linear regression analysis data were used for the regression line in the range of 0.25–0.85 and 0.10–1.00 μg band−1 for LOR and DES, respectively. The method was successfully applied to the analysis of the two drugs in their commercial tablets, capsules, and syrups, and the results were in good agreement with those obtained with the comparison method. The proposed method is specific for the simultaneous determination of loratadine and desloratadine in the presence of other co-formulated drugs, such as pseudoephedrine. The method is...

Development and validation of TLC-densitometry method for simultaneous determination of telmisartan and amlodipine besylate in bulk and tablets

A rapid, simple, and selective high performance thin layer chromatographic method was developed and validated for simultaneous estimation of telmisartan and amlodipine besylate in pharmaceutical dosage forms. The method employed TLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent system comprised: tetrahydrofuran: dichloroethane: methanol: ammonia solution (6.0:2.0:1.0:0.4 v/v). This system was found to give compact spots for both telmisartan (Rf value of 0.22 ± 0.02) and amlodipine besylate (Rf value of 0.45 ± 0.02). Spectrodensitometric scanning-integration was performed at a wavelength of 326 nm. The polynomial regression data for the calibration plots showed good linear relationship with r2 = 0.9993 in the concentration range of 1,200–7,200 ng for telmisartan and 400–1,400 ng for amlodipine besylate with r2 = 0.9996. The method was validated for precision, accuracy, ruggedness, and recovery. The minimum detectable amounts were found to be 149.41 ng and 53.07 ng for telmisartan and amlodipine besylate, respectively. The limits of quantitation were found to be 452.78 ng for telmisartan and 160.83 ng for amlodipine besylate. Statistical analysis proves that the method is reproducible and selective for the simultaneous estimation of telmisartan and amlodipine besylate. Key words: Amlodipine besylate, HPTLC, simultaneous determination, telmisartan.

Analysis of levofloxacin in pharmaceutical preparations by high performance thin layer chromatography

AbstractA new simple, precise, rapid, and selective high‐performance thin layer chromatography (HPTLC) method has been developed for the analysis of levofloxacin in pharmaceutical formulations. The method uses lamotrigine as an internal standard. The stationary phase was silica gel 60F254 prewashed with methanol; water‐methanol‐n‐butanol‐ammonia solution 5 + 5 + 5 + 0.4 (v/v) was used as mobile phase. Detection and quantification were performed densitometrically at λ = 298 nm. The linear range of the analysis was 0.8–3.0 μg and the percentage recovery was 99.90%.

HPTLC method to study skin permeation of acylovir

A new simple, rapid and selective high performance thin layer chromatography (HPTLC) method is developed for the quantitation of acyclovir during in vitro skin permeation studies. Separation of guinea pig skin proteins and acyclovir was achieved by employing a mobile phase consisting of chloroform–methanol–ammonia (15:9:4, v/v/v) on precoated silica gel 60F254 aluminum plates. Densitometnic analysis was carried out at 255 nm. The limit of detection and quantification were 30 and 50 ng, respectively. The calibration curve was linear in the range of 10–20 μg/ml ( r=0.9965). The relative standard deviation for a sample of concentration 100 μg/ml were 1.15 and 2.85 for system and method precision, respectively. Intraday and interday variation studies gave an average 0.763 and 0.463% relative standard deviation for the three levels tested. Average recoveries of 101.8 and 100.1% were recorded for two marketed preparations studied. The method was employed to optirmize topical liposomal gel formulation of acyclovir on basis of maximum skin permeation.