Sensitive High-performance Thin-layer Chromatographic Method Research Articles

A Simple and Sensitive HPTLC Method for Simultaneous Determination of Metformin Hydrochloride and Sitagliptin Phosphate in Tablet Dosage Form

A simple, rapid, and precise high‐performance thin‐layer chromatographic (HPTLC) method for simultaneous estimation of two antidiabetic drugs, metformin hydrochloride and sitagliptin phosphate, in tablet dosage form has been developed and validated. Chromatography was performed on silica gel 60 F254 plates with butanol : water : glacial acetic acid (6 : 2 : 2, v/v/v) as mobile phase. This system gave a good resolution for metformin hydrochloride (Rf value of 0.35 ± 0.01) and sitagliptin phosphate (Rf value of 0.75 ± 0.01). Detection and quantification were carried out at 227 nm. The linear regression data for the calibration plot showed a good relationship with r = 0.9995 and 0.9991 for metformin hydrochloride and sitagliptin phosphate, respectively. The method was validated for precision and recovery. The limits of detection and quantification were 13.05 and 39.56 ng/μL for metformin hydrochloride and 2.65 and 8.03 ng/μL for sitagliptin phosphate, respectively. The amounts of the drugs in the marketed formulation were 99.86% and 98.91% for metformin hydrochloride and sitagliptin phosphate, respectively.

A simple, rapid, and sensitive HPTLC method for the estimation of clarithromycin: Application to single dose clinical study

Clarithromycin used for the treatment of respiratory tract infection. Anew alternative simple, rapid, sensitive, and selective high-performance thin-layer chromatography (HPTLC) method was developed and validated for the estimation of clarithromycin in plasma. HPTLC analysis was performed on pre-coated silica gel 60F254 plates. Separation was carried out using ethyl acetate-methanol-15% ammonium acetate (pH 10.6) as solvent system. Densitometry measurements were performed in absorbance mode at 506 nm after derivatising with xanthydrol solution. The RF of the drug was found to be 0.62. The method was linear over the range of 0.1–3.0 μg mL−1 (r2 = 0.9974). The mean extraction recoveries were over 85%. The intra-day and inter-day precision (%CV) of the assay was in the range of 0.79–4.55%. The accuracy was found to be above 95%. The method was successfully applied to a pharmacokinetic study of clarithromycin 250 mg tablet in healthy human male volunteers.

A simple and sensitive HPTLC method for quantitative analysis of darunavir ethanolate tablets

A quantitative high-performance thin-layer chromatography (HPTLC) method for determination of darunavir ethanolate (DRV) in tablets has been established and validated. DRV from the formulations was separated and identified on silica gel 60 F254 HPTLC plates with toluene-ethyl acetate-methanol 7.0:2.0:1.0 (ν/ν) as mobile phase. The plates were developed to a distance of 8 cm. Quantification was performed at λ = 267 nm. Well-resolved bands were obtained for DRV. The method was validated for specificity, precision, robustness, and accuracy. The calibration plot for DRV standard was linear in the range 250–1750 ng per band with r = 0.9994, slope = 0.4253, and intercept = 44.81. The limits of detection and quantification were 15.28 and 45.84 ng per band, respectively. The method is selective, sensitive, and specific, with potential application in pharmaceutical analysis.

A simple and sensitive HPTLC method for simultaneous analysis of domperidone and paracetamol in tablet dosage forms

A simple, precise, rapid, selective, and economic high-performance thin-layer chromatographic (HPTLC) method has been established for simultaneous analysis of domperidone (DMP) and paracetamol (PAR) in tablet dosage forms. The chromatographic separation was performed on precoated silica gel 60 GF 254 plates with acetone-toluene-methanol 4:4:2 ( v/v ) as mobile phase. The plates were developed to a distance of 8.0 cm at ambient temperature. The developed plates were scanned and quantified at their single wavelength of maximum absorption at approximately 285 and 248 nm for domperidone and paracetamol, respectively. Experimental conditions such as band size, chamber saturation time, migration of solvent front, slit width, etc. were critically studied and the optimum conditions were selected. The drugs were satisfactorily resolved with R F 0.52 ± 0.02 for domperidone and 0.74 ± 0.02 for paracetamol. The method was validated for linearity, accuracy, precision, and specificity. The calibration plot was linear b...

A Simple and Sensitive HPTLC Method for Simultaneous Determination of Abacavir Sulphate and Lamivudine in Tablet Dosage Form

A simple, sensitive, precise and rapid high performance thin layer chromatographic (HPTLC) method has been developed for the quantitative estimation of abacavir sulphate and lamivudine in tablet dosage form. Abacavir sulphate and lamivudine were chromatographed on silica Gel 60 GF-254 TLC plate. The solvent system selected was methanol:acetone:n-butyl acetate in the ratio of 1:1:2 (v/v/v) and scanning was carried out in the absorbance mode at 284 nm. The spots were found compact for abacavir sulphate with Rf value of 0.58 and for lamivudine with Rf value of 0.35. Linearity of abacavir sulphate and lamivudine were in the range of 240-1200 ng/spot and 120-600 ng/spot, respectively with acceptable value of correlation coefficient. The method was validated for accuracy, precision, robustness and recovery studies. The limit of detection and quantification for abacavir sulphate were obtained 0.691 and 2.093 ng/spot, respectively, while for lamivudine 1.114 and 3.376 ng/spot, respectively. The suitability of this HPTLC method for quantitative determination of abacavir sulphate and lamivudine was proved by validation in accordance with the ICH guidelines. The developed method can be successfully applied in the routine analysis of commercial pharmaceutical tablets.

Rapid HPTLC analysis of oxcarbazepine in human plasma

A sensitive high-performance thin-layer chromatographic method has been developed for analysis of oxcarbazepine (OXC) in human plasma. This paper describes extraction of OXC from spiked human plasma by protein precipitation with acetonitrile and quantification using normal-phase HPTLC. The optimized mobile phase was ethyl acetate-toluene-methanol 7.0:2.0:1.0 ( v/v ). The detection was performed densitometrically at 254 nm by UV absorption. The detection limit of OXP is 2 ng and the limit of quantification is 10 ng ( n = 6). A good correlation coefficient ( r 2 = 0.99) was obtained for amounts of OXC in the range 10 to 3000 ng. Recovery was 75 to 80%. The internal standard used was chlorzoxazone, which was well resolved from OXC under the conditions specified. Analysis of OXP in human plasma was successfully performed without interference from endogenous materials.

HPTLC analysis of octachlorodipropyl ether in insecticide formulations

A simple, rapid, and sensitive high-performance thin-layer chromatographic method for analysis of octachlorodipropyl ether (OCDPE) in insecticide formulations has been established and validated. Known amounts of analytical grade OCDPE and its emulsifiable concentrate (EC) or wettable powder (WP) formulations were characterized by HPTLC on silica gel with toluene-acetic acidwater 20:20:1 ( v/v ) as mobile phase; detection was by spraying with silver nitrate-2 m alcoholic potassium hydroxide as chromogenic reagent and exposure to UV light. The plates were evaluated densitometrically at 399 nm. The results indicated that the calibration plot for OCDPE was logarithmic in the range 0.2–5.0 μg per band, and the correlation coefficient for the calibration equation in this range was 0.99. Recoveries from laboratory-prepared test EC and WP formulations using this method were 98.5–103.9% and 95.3–104.3%, respectively, and the respective RSDs were 3.39–4.89% and 2.92–5.33%. The accuracy and precision of the method w...

HPTLC method for quantification of apigenin in the dried root powder ofGmelina arboreaLinn.

A sensitive high-performance thin-layer chromatographic method has been established for quantification of apigenin in dried root powder from Gmelina arborea Linn. Chromatographic separation was performed on silica gel plates with chloroform-acetone-formic acid, 7.6:1.6:0.8 ( v / v ), as mobile phase. The plates were scanned densitometrically at 340 nm. The method was validated for precision and accuracy. The relative standard deviation for instrumental precision, intra-assay precision, and intermediate precision was <2%. Percentage recovery was 98.87.

Rapid and sensitive HPTLC method for determination of epalrestat in human plasma

Epalrestat is a new drug for diabetic neuropathy. A new, simple, rapid very sensitive, and accurate high performance thin-layer chromatographic (HPTLC) method has been developed and validated for estimation of epalrestat in plasma. HPTLC was performed on silica gel 60F 254 plates with ethyl acetate-toluene-acetic acid, 60 + 40 + 2 ( v / v ), as mobile phase. Nitrofurantoin was used as internal standard. Densitometric scanning was performed in absorbance mode at λ = 390 nm. The R F values of the internal standard and drug were 0.4 and 0.6, respectively. The response was a linear function of concentration over the range 0.1–6.0 μg mL −1 ( r 2 = 0.9974). Mean extraction recovery was > 63%. Intra-day and inter-day precision (% CV) of the assay were in the range 1.62–4.68% and accuracy was > 95%. This method can be applied to pharmacokinetic and bioequivalence studies.

HPTLC determination of apigenin in some Iranian liquid products ofMatricaria chamomillaL.

A simple, rapid, and sensitive high-performance thin-layer chromatographic method has been developed for determination of apigenin in liquid products of Matricaria chamomilla L. (syn. Chamomilla recutita L.). Samples were applied directly to silica gel 60F 254 HPTLC plates as stationary phase. Plates were developed to a distance of 90 mm in a glass twin-trough chamber with toluene-methanol, 10 + 2 ( v/v ), as mobile phase. The plates were scanned densitometrically at λ = 343 nm. The method was validated for selectivity, repeatability, and accuracy. The repeatability of the method was excellent with a mean RSD of 2.2%. The accuracy was checked by studying recovery at two levels and average recovery was 101.4%. The method was confirmed to be suitable for determination of apigenin in Matricaria chamomilla liquid products.

HPTLC determination of imidacloprid, fenitrothion and parathion in Chinese cabbage

A simple, rapid, and sensitive high-performance thin-layer chromatographic method has been developed and validated for analysis of residues of imidacloprid, fenitrothion, and parathion in Chinese cabbage. The sample was extracted by sonication in an ultrasonic water bath with acetone-petroleum ether, 5:3 ( v / v ), and the extract was directly applied, as bands, to glass-backed silica gel 60F 254 HPTLC plates. The plates were developed with hexane-acetone, 7 + 3 ( v / v ), in an unsaturated glass twin-trough Camag chamber. Evaluation of the developed HPTLC plates was performed densitometrically with a Camag TLC Scanner 3 controlled by an external PC running Wincats software (Version 1.1.2). The results indicated that the detection limits of imidacloprid ( R F = 0.10), fenitrothion ( R F = 0.59), and parathion ( R F = 0.70) were 5.0 × 10 −9 g, 2.0 × 10 −8 g, 1.0 × 10 −8 g, respectively. Recoveries of the pesticides from Chinese cabbage by use of this analytical method were 80.04–85.22%, and RSD were 4.18–1...

A convenient and high throughput HPTLC method for determination of progesterone in release media of silicon-based controlled-release drug-delivery systems

A simple, rapid, and sensitive high-performance thin-layer chromatographic method has been developed for determination of progesterone in release media of silicon-based controlled-release drug-delivery systems. Samples were applied directly to silica gel 60F 254 as stationary phase and plates were developed to a distance of 60 mm, in an automatic multiple development chamber, with toluene-2-propanol, 9 + 1 ( v/v ), as mobile phase, without chamber saturation and with 10 min drying time. This system resulted in dense, compact zones for the analyte at R F 0.56. Regression data for the calibration plots were indicative of a good linear relationship ( R 2 = 0.998) over the concentration range 25–125 ng/zone. Limits of quantitation and detection were 25 and 5 ng/zone, respectively. The mean recovery of progesterone from samples was 100.77%. The mean between-day accuracy for analysis of progesterone in the range 27.70–144.89 ng/zone, including three parallel measurements, was 100.34%.

Development of a sensitive high-performance thin-layer chromatography method for estimation of ranitidine in urine and its application for bioequivalence decision for ranitidine tablet formulations

A sensitive and simple HPTLC method was developed for estimation of ranitidine in human urine. The drug was extracted from urine after basification using dichloromethane. Dichloromethane extract was spotted on silica gel 60 F 254 TLC plate and was developed in a mixture of ethyl acetate–methanol–ammonia (35:10:5 v/v) as the mobile phase and scanned at 320 nm. The R F value obtained for the drug was 0.67±0.03. The method was validated in terms of linearity (50–400 ng/spot), precision and accuracy. The average recovery of ranitidine from urine was 89.35%. The proposed method was applied to evaluate bioequivalence of two marketed ranitidine tablet formulations (150 mg, Formulation 1 and Formulation 2) using a crossover design by comparing urinary excretion data for unchanged ranitidine in six healthy volunteers. Various pharmacokinetic parameters like peak excretion rate [(dAU/d t) max], time for peak excretion rate ( t max), AUC 0-24, AUC 0-∞, cumulative amount excreted were calculated for both formulations and subjected to statistical analysis. The relative bioavailability of Formulation 2 with respect to Formulation 1 was 93.76 and 95.31% on the basis of AUC 0-24 and cumulative amount excreted, respectively. Statistical comparison of various pharmacokinetic parameters indicated that the two ranitidine tablet formulations are bioequivalent.

Sensitive high-performance thin-layer chromatographic method for the estimation of diospyrin, a tumour inhibitory agent from the stem bark of Diospyros montana Roxb.

Diospyrin, a tumour inhibitory agent from the stem bark of Diospyros montana was isolated and characterised. A sensitive high-performance thin-layer chromatographic (HPTLC) method was developed for the estimation of diospyrin. The method was validated for precision (intra- and inter-day), repeatability and accuracy. The method was found to be precise, with the RSDs for intra-day in the range of 0.72–1.85% and RSDs for inter-day in the range of 1.06–2.95%, for different concentrations. Instrumental precision and repeatability of the method were found to be 0.086 and 0.937 (% CV), respectively. Accuracy of the method was checked by performing the recovery study at two levels and average percentage recovery was found to be 97.87%. The developed HPTLC method was adopted for the estimation of diospyrin content of the stem bark of D. montana from different regions, which varied from 0.35 to 0.47% (w/w) in the samples.

Sensitive high-performance thin-layer chromatography method for detection and determination of ranitidine in plasma samples

A high-performance thin-layer chromatographic procedure has been developed for the determination of ranitidine, a H 2-receptor antagonist, in plasma. The detection and quantification were performed without using internal standards. A single-stage extraction procedure was followed for extracting ranitidine from plasma, and a known amount of the extract was spotted on precoated silica gel F254 plates. Ranitidine was quantified using a Shimadzu CS930 dual-wavelength TLC scanner. The method provides a direct estimate of total ranitidine present in the plasma.

10 - Cell Separation and Surface Markers

To select the best organic solvent and ideal extraction technique for isolation and quantitative estimation of ursolic acid from the leaves of Lantana camara (L. camara) by high performance thin layer chromatography (HPTLC). A sensitive, fast and reproducible HPTLC method was developed and validated for estimation of ursolic acid from the leaves of L. camara. Different leaves extracts of L. camara were prepared through different extraction methods (maceration, reflux, soxhlet and ultrasound assisted extraction technique) by using four different solvents of varying polarity such as ethanol, methanol, acetone and chloroform. The maximum % extract yield was obtained in methanol, so we select methonolic extract for HPTLC evaluation. The calibration plot of standard ursolic acid with respect to peak area showed an tremendous linear relationship in the concentration range of 1000–4000 ng/spot with a correlation coefficient r2 = 0.994. The separation was completed on precoated silica gel aluminium plates 60F254 using Toluene: Acetone: Formic Acid (7.8:2.2:0.15) as mobile phase which gives a compact spot of ursolic acid (Rf value 0.49). For the proposed method (Limit of Detection) LOD (561.85 ng/spot), (Limit of Quantification) LOQ (1702.58 ng/spt), recovery of drug (99.02–101.43%) and precision (0.4303–2.1576% RSD) was found to be satisfactory. It can be concluded on the basis of HPTLC results that methanol was found to be the best solvent and Soxhlet was found to be an ideal extraction technique for the isolation of ursolic acid from L. camara leaves for routine analysis.