Giloy Tulsi tablet is an Ayurvedic preparation containing Tinospora cordifolia (Giloy) and Ocimum sanctum (Tulsi) and it is recommended for boosting the body’s immune response. This research work is about the marker-based standardization of this Ayurvedic preparation using high-performance thin-layer chromatography method. Standardization is based on the determination and quantification of the phytoconstituents berberine and ursolic acid present in Giloy Tulsi tablets. Separation was performed on pre-coated silica gel 60 F254 plate as the stationary phase, with chloroform‒acetone‒formic acid (6:3.5:0.5, V/V) as the mobile phase. Identification and quantification were conducted densitometrically at 330 nm. The developed method resulted in good quality peak shape and enabled high-quality resolution of biomarkers. The RF value for berberine (0.46 ± 0.02) and for ursolic acid (0.68 ± 0.02) in both reference standard and formulation were found to be comparable. The method was validated for specificity, linearity, limit of detection, limit of quantification, intra-day and inter-day precisions, accuracy, and robustness. The limit of detection values were 91 and 153 ng/band for berberine and ursolic acid, respectively. The limit of quantification values were 175 and 465 ng/band for berberine and ursolic acid, respectively. Regression analysis of the calibration data revealed a good linear relationship between peak area response and concentration in the range 200‒1000 ng/band for berberine (r2 = 0.995) and 500‒2500 ng/band for ursolic acid (r2 = 0.9968). The accuracy of the method, determined by measurement of recovery at three different levels, was in the range 98‒102% for both markers. These results are indication of reliability, reproducibility, accuracy, and precision of the method.
相似文献A novel high-performance thin-layer chromatographic (HPTLC) analytical method has been developed and optimized for the quantification of quetiapine fumarate (QF) and its two genotoxic impurities in drug substance and drug product. The desired separation was achieved on 60F254 pre-coated HPTLC plates using combination of green solvents, ethyl acetate‒ethanol‒n-heptane (5:1:4, V/V) as developing solvents. The detection wavelength used for quantification was 229 nm. QF and its two related genotoxic impurities, namely, 2-chloroaniline and 2-aminodiphenylsulfide, were well resolved from one another with retention factor values of 0.13 ± 0.02, 0.57 ± 0.02 and 0.76 ± 0.02, respectively. The optimized method was validated according to the guidelines laid down by the International Council for Harmonisation. The linearity was determined in the range of 100–600 ng/spot for QF and 10‒60 ng/spot for its two related genotoxic impurities; R2 ≥ 0.993. The method exhibited precision along with good accuracy, where 0.51, 0.86 and 1.86. The percentage recoveries obtained for 2-chloroaniline and 2-aminodiphenylsulfide were 99.04‒101.04%. The developed method can be successfully used for the analysis of drug samples.
相似文献A validated high-performance thin-layer chromatography (HPTLC) method was developed for the simultaneous quantification of oleanolic acid, β-sitosterol and lupeol in the bulb of Urginea indica Kunth. Separation of metabolites was done in mobile phase using toluene‒ethyl acetate‒methanol‒acetone (7:2:0.2:0.2, V/V) and quantification was done after derivatization by dipping in aninsaldehyde‒sulphuric acid; densitometric scan was performed at 530 nm. The proposed method for quantification was linearly calibrated in the range of 200‒1000 ng/spot for oleanolic acid and β-sitosterol; 100‒500 ng/spot for lupeol, and it was found specific and repeatable. The RF values were found at 0.44 ± 0.03, 0.55 ± 0.05 and 0.68 ± 0.08, limit of detection and limit of quantification were 1.045, 0.524, 0.525 µg/spot and 3.167, 1.588, 1.592 µg/spot for oleanolic acid, β-sitosterol and lupeol, respectively. Precision and recovery study for sample and standards were within the limit of the International Council for Harmonization guidelines. Oleanolic acid, β-sitosterol and lupeol were found to be 0.113%, 0.105% and 0.036%, respectively, in methanolic extract of plant on dry weight basis. This study will help in checking routine quality control of herbal drugs as well as herbal formulations containing U. indica.
相似文献The lipase inhibitory activities of four main components from the rhizomes of Alisma orientale (Sam.) Juz. were evaluated by an in situ high-performance thin-layer chromatography (HPTLC)‒bioautographic assay taking orlistat as control standard. The order of relative activity was alisol B 23-acetate > alisol B > alisol A > alisol C 23-acetate. With that, an accurate, efficient and sustainable HPTLC method was developed to simultaneously determine the four lipase inhibitors from the methanolic extracts of Alismatis Rhizoma (AR). The method was carried out on HPTLC glassed plates (20 × 10 cm) coated with silica gel 60 F254 (0.2 mm thickness) using a mixture of cyclohexane and ethyl acetate (1:1, V/V) as the mobile phase. The RF values found for alisol B 23-acetate, alisol C 23-acetate, alisol B and alisol A were 0.62, 0.42, 0.28 and 0.09, respectively. The method was validated for specificity, linear range, precision, stability, and recovery. The results determined by scanning densitometry showed no significant difference to the results obtained by HPLC. The developed method was verified to be trustworthy for the evaluation of quality markers in AR.
相似文献Himalaya PartySmart capsule is a polyherbal formulation recommended for its liver-protective properties. As the formulation contains extracts of six different herbs, a large number of markers are present in the same. This research work reports the standardization of Himalaya PartySmart capsule using andrographolide and catechin as therapeutic phytoconstituents to assess its quality and efficacy. A specific, sensitive, precise, and accurate high-performance thin-layer chromatography (HPTLC) method has been developed for the quantitative estimation of andrographolide and catechin. Separation was performed on TLC silica gel 60 F254 aluminum plates as the stationary phase using chloroform‒acetone‒formic acid (7:3:0.5, V/V) as the mobile phase with densitometric detection at 259 nm. The developed method was validated as per the recommendations of the International Council for Harmonisation (ICH) Q2(R1) guideline. Each marker phytoconstituent showed a good linear relationship with an average correlation coefficient (r2) = 0.99 in the concentration range studied. The proposed method was found to be specific, precise, and accurate with recovery within the range of 95‒105% and hence can be used for the routine analysis of PartySmart capsule formulation.
相似文献A simple, precise, rapid and accurate high-performance thin-layer chromatographic method was developed and validated for the simultaneous estimation of emtricitabine (EMT) and tenofovir alafenamide fumarate (TAF) in combined dosage forms and forced degradation studies were carried out as per the International Council for Harmonisation (ICH) guidelines. The stationary phase used was Merck TLC silica gel 60 F254 aluminum plates. The mobile phase used was ethyl acetate‒n-hexane‒methanol‒ammonia solution (4:4:2:0.2, V/V). Densitometric evaluation was carried out at 260 nm, EMT and TAF. The concentration ranges used for the study are 400–2000 ng/band for EMT, 50–250 ng/band for TAF. EMT and TAF gave well defined and sharp peaks at retardation factor 0.43 and 0.56, respectively. The stability study showed that samples degraded with acid, base, hydrogen peroxide and light gave well separated peaks of EMT and TAF as well as some additional peaks at different retardation factor values. The proposed method is simple, suitable, accurate and stable in accordance with the ICH guidelines.
相似文献According to the International Council for Harmonisation (ICH) Q2 (R1) guideline, a sensitive, precise, accurate and robust high-performance thin-layer chromatographic (HPTLC) method was developed and validated for the simultaneous quantification of a newer combination of brexpiprazole (BREX) and sertraline HCl (SERT) in bulk and synthetic mixture. Stationary phase selected was pre-coated silica gel aluminum plate 60 F254, and n-propanol‒hexane‒toluene‒triethylamine (7:2:1:0.1, V/V) was used as developing mobile phase. An appreciable absorbance shows at 254 nm, therefore the common detection wavelength was selected for the simultaneous quantification of BREX and SERT. The method was validated for different parameters: linearity, precision, accuracy, robustness, limit of detection and limit of quantification as per ICH guideline. The correlation coefficients (r2) for BREX and SERT were found to be 0.9940 and 0.9911, respectively. The mean of percentage recoveries for BREX and SERT were found to be 99.40–102.10% and 99.52–101.05%, respectively. The proposed HPTLC method has potential application for the quantification of BREX and SERT simultaneously in bulk and synthetic mixture both qualitatively and quantitatively.
相似文献A new, simple, precise, accurate and selective high-performance thin-layer chromatographic (HPTLC) method has been developed and validated for the simultaneous determination of ledipasvir and sofosbuvir in their tablet dosage form. Chromatographic separation was carried out on Merck TLC aluminum sheets of silica gel 60 F254 using ethyl acetate:hexane:methanol in the ratio of 8:1.25:0.75 (% v/v) as the mobile phase followed by densitometric measurement at 256 nm. The method was validated in terms of linearity, accuracy, precision, limit of detection, limit of quantification and specificity in accordance with the International Conference on Harmonization (ICH) guidelines. The calibration curve was found to be linear between 60 to 1980 and 45 to 3600 ng/band for ledipasvir and sofosbuvir, respectively, with significantly high value of regression coefficient (r2 > 0.9999) with linear and homoscedastic residuals. The limits of detection and quantification were found to be 16.5 and 50 ng/band, respectively, for ledipasvir and 13 and 39.5 ng/band, respectively, for sofosbuvir. Comparative study was performed between the developed HPTLC method and the reported high-performance liquid chromatography (HPLC) method. The quantitative results of the two analytical methods did not show statistically significant difference, whereas the developed HPTLC method is both time- and cost-effective.
相似文献A simple, selective, precise, rapid and accurate stability-indicating high-performance thin-layer chromatography (HPTLC) method was developed and validated for the estimation of dapagliflozin and metformin in tablet dosage form. In this work, methanol–ethyl acetate–ammonium acetate (6:4:0.1, V/V) as the mobile phase and aluminum-backed TLC plates pre-coated with 250 µm layer of silica gel 60F254 as the stationary phase were used for the estimation of dapagliflozin and metformin. The wavelength selected for detection was 220 nm. The linearity range was found to be 20–100 ng/spot (r2 = 0.9985) for dapagliflozin and 500–2500 ng/spot (r2 = 0.9984) for metformin. Validation of the developed method was performed as per the International Council for Harmonisation (ICH) guidelines. Stress testing of dapagliflozin and metformin was performed under acidic, alkaline, oxidative, photolytic and dry-heat degradation conditions. The chromatographic conditions successfully resolved dapagliflozin and metformin from their degradation products, formed under various stress conditions. From stress testing, dapagliflozin was found to be significantly degrading under acidic, alkaline, oxidative, photolytic and dry-heat degradation conditions, while metformin was found to be significantly degrading in acidic and alkaline degradation conditions and stable under oxidative, photolytic and dry-heat degradation conditions. Tablet dosage form of dapagliflozin and metformin was analyzed by the developed method.
相似文献We present a densitometric quantification method for triclosan in toothpaste, separated by high-performance thin-layer chromatography (HPTLC) and using a 48-bit flatbed scanner as the detection system. The sample was band-wise applied to HPTLC plates (10 × 20 cm), with fluorescent dye, Merck, Germany (1.05554). The plates were developed in a vertical developing chamber with 20 min of chamber saturation over 70 mm, using n-heptane–methyl tert-butyl ether–acetic acid (92:8:0.1, V/V) as solvent. The RF value of triclosan is hRF = 22.4, and quantification is based on direct measurements using an inexpensive 48-bit flatbed scanner for color measurements (in red, green, and blue) after plate staining with 2,6-dichloroquinone-4-chloroimide (Gibbs' reagent). Evaluation of the red channel makes the measurements of triclosan very specific. For linearization, an extended Kubelka–Munk expression was used for data transformation. The range of linearity covers more than two orders of magnitude and is between 91 and 1000 ng. The separation method is inexpensive, fast and reliable.
相似文献A novel, simple, precise, specific, accurate high-performance thin-layer chromatography (HPTLC) method was developed and validated for the estimation of bromfenac in ophthalmic solution. Diclofenac sodium was used as an internal standard (IS) because of its structural resemblance with bromfenac to develop a more accurate and precise method. Silica gel 60 F254 HPTLC plates were used to separate bromfenac from the formulation with a mobile phase consisting of toluene-ethyl acetate-glacial acetic acid (65:35:0.2, V/V). Densitometric scanning was performed at 274 nm after the HPTLC plates were air-dried. Well-resolved bands and good peak shapes were obtained for both bromfenac and diclofenac sodium, with retention factor (RF) values of 0.28 and 0.44, respectively. The proposed method was validated as per International Council for Harmonisation Q2 (R1) guidelines for specificity, precision, robustness, accuracy, and recovery. The drug shows linearity in the concentration range of 60‒270 ng/band and the correlation coefficient was found to be 0.999. The mean percent recovery of bromfenac was found to be 100.7%. The limit of detection and limit of quantification values for bromfenac were found to be 7.4 ng/band and 22.5 ng/band, respectively. The method was found to be novel since no HPTLC methods have yet been reported for the estimation of bromfenac. The developed method was successfully applied for the quantitative analysis of the drug in the ophthalmic formulation.
相似文献No high-performance thin-layer chromatography (HPTLC) techniques have been established for the determination of tedizolid phosphate (TDZP) in pharmaceutical products or physiological fluids. Therefore, a rapid and highly sensitive stability-indicating HPTLC technique has been developed for the determination of TDZP in commercial formulations with a classical univariate calibration. The HPTLC‒densitometry analysis of TDZP was carried out via chloroform‒methanol (90:10, V/V) mobile phase. The determination of TDZP was performed at the wavelength of 300 nm. The proposed HPTLC technique was linear in the range of 10‒2000 ng band‒1. In addition, the method was found to be highly accurate (% recovery = 98.53‒101.74%), precise (%CV = 0.67‒0.91%), robust (%CV = 0.83‒0.86%), highly sensitive (LOD = 3.41 ng band‒1, LOQ = 10.23 ng band‒1) for the determination of TDZP. The proposed technique was also able to detect TDZP in the presence of its degradation products under various stress conditions and it can be considered as a stability-indicating method. The proposed HPTLC technique was applied for the analysis of TDZP in its commercial formulations. The TDZP contents of commercial tablets and injection were determined as 98.41% and 101.23%, respectively. These results suggested that the proposed HPTLC technique can be applied for the routine analysis of TDZP in its commercial products and newly established formulations.
相似文献Mucuna pruriens Linn. one of the popular and important medicinal plants of India is a constituent of more than 200 indigenous drug formulations. β-Sitosterol is one of the most prevalent phytosterols which is ubiquitous throughout the plant kingdom. A sensitive, selective and precise thin-layer chromatographic method has been developed and validated for the analysis of β-sitosterol in Mucuna pruriens roots. Separation and quantification was achieved by TLC using ternary mobile phase of toluene: chloroform; methanol (4:4:1 v/v) (R F 0.55) on precoated silica gel 60F254 aluminium plates and densitometric determination was carried out after derivatization with anisaldehyde-sulphuric acid reagent in reflection/absorption mode at 527 nm. The calibration curve was linear in the concentration range of 100–600 ng spot−1. The method was validated for precision, repeatability and accuracy. The proposed method was found to be simple, precise, specific, sensitive and accurate for the quantification of β-sitosterol.
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