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Two validated, simple and precise densitometric high-performance thin-layer chromatography (HPTLC) quantification methods were proposed for both qualitative and quantitative estimation of oleuropein in Olea europaea leaves and a pharmaceutical product utilizing normal-phase and reversed-phase silica gel TLC plates. In method I, 10 × 20 cm glass plates coated with 0.2 mm thin layers of normal-phase silica gel 60 containing F254 (E-Merck, Germany) and a mixture of ethyl acetate‒methanol‒water (8:1:0.5, V/V) were used as the stationary and the mobile phase, respectively. Method II utilized 10 × 20 cm glass-backed plates supporting 0.2 mm layers of RP-18 silica gel 60 containing F254 (E-Merck, Germany) as the stationary phase and green solvents mixture composed of ethanol‒water (5.5:4.5, V/V) as the mobile phase. The two methods resulted in sharp, symmetrical, well-resolved peaks at RF values of 0.47 ± 0.02 and 0.78 ± 0.03 with linearity ranges 200‒1400 ng/spot (r2 = 0.9994) and 200‒1400 ng/spot (r2 = 0.9996) for method I and method II, respectively. Spots corresponding to oleuropein were scanned at 200 nm. The two methods complied with the ICH guidelines for validation. Due to simplicity, low cost and short analysis time, the methods can be good alternatives for the quality control of different products containing olive leaves extract or pure oleuropein.
相似文献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 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A sensitive, reliable and rapid high-performance thin-layer chromatography (HPTLC) method for the determination of arctiin and arctigenin in Arctium tomentosum Mill. was established. A. tomentosum Mill. extract was used for chromatographic analysis. The ratio of chloroform and methanol was 48:5 as mobile phase. Temperature is 20–23 °C and humidity is less than 30%. The scanning wavelength is 280 nm. The results showed that arctiin had a good linear relationship in the range of 0.5315–5.8465 μg, r = 0.9982; arctigenin had a good linear relationship in the range of 0.5654–6.2194 μg, r = 0.9951. Precision analysis showed that the RSD < 3.0%. The stability study showed that the sample was stable within 24 h at room temperature, RSD < 2.0%. The average recoveries were 103.07 ± 1.57% and 98.55 ± 2.71%, respectively. The antioxidant activity of Arctium tomentosum Mill. was also identified. The results showed that the antioxidant component identified by thin-layer chromatography–1, 1-diphenyl-2-trinitrophenylhydrazine (TLC-DPPH) was arctigenin not arctiin. The proposed HPTLC is a simple and accurate method for the qualitative and quantitative analysis of arctiin and arctigenin in Arctium tomentosum Mill. from different areas.
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Cucumis sativus L. of the Cucurbitaceae family, commonly known as cucumber, is commercially cultivated worldwide. The major phytoconstituents present in the Cucurbitaceae family are different curcurbitacins, principally cucurbitacin E. The content of cucurbitacin E differs within the species or cultivars due to factors like genetic variation and geographical location. The present study reports a simple and rapid quantitative analysis of cucurbitacin E in 5 different C. sativus cultivars by a validated high-performance thin-layer chromatography (HPTLC) method. The mobile phase contained petroleum ether, ethyl acetate and formic acid in the ratio of 40:60:0.5 (V/V). Cucurbitacin E was analyzed densitometrically and the absorbance wavelength was 254 nm. The method showed RF spot = 0.79 ± 0.06, corresponding to cucurbitacin E in various samples. The calibration curve of standard cucurbitacin E showed good linear relationship in the concentration range of 2‒10 µg/spot with a correlation coefficient (r) > 0.99. The HPTLC method was validated in terms of sensitivity, linearity, accuracy, precision, and specificity as per the International Conference on Harmonization (ICH) guidelines. The present study revealed that the content of cucurbitacin E differs among the C. sativus cultivars. This method may be beneficial for addressing the quality-related aspects of C. sativus for food and pharmaceutical preparation.
相似文献High-performance thin-layer chromatography‒mass spectrometry (HPTLC‒MS) method was developed for the estimation of epimers (+)-catechin (CA) and (‒)-epicatechin (ECA) in Onosma bracteatum Wall. Resolving these epimers is challenging and so method optimization was done for the selection of the stationary phase and the mobile phase to achieve their coherent separation. To further increase the reliability of the obtained densitometric results, HPTLC–MS analysis was performed. The genus Onosma L. is a species-rich genus that exhibits complex patterns of morphological and karyological diversity, and highly debatable taxonomic approaches. Thus, many similar species are described based on morphological differences and often quite ambiguous. To facilitate the identification of O. bracteatum, separation was achieved using pre-coated silica gel 60 F254 HPTLC plate as the stationary phase and a mixture of diisopropyl ether–ethyl acetate–formic acid (9.0:0.2:0.7, V/V) as the mobile phase for the separation of epimers CA and ECA. Sample preparation, mobile phase selection and optimization were given importance to manage good resolution (RF) of these markers. Flavan-3-ols CA and ECA were identified and confirmed on the basis of RF and in situ UV and MS overlaid spectra with respective standards. The method was validated for linearity, inter-day precision, intra-day precision, repeatability, accuracy, specificity, limit of detection, and limit of quantification. The average recoveries for epimers CA and ECA from ethyl acetate extract fraction (MEF) were found 98.86 and 99.03% indicating the good reproducibility for each marker. The proposed validated HPTLC method is simple, accurate and reproducible and is the first report on the separation and quantification of the epimers CA and ECA in O. bracteatum using HPTLC–MS.
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Qternmet XR® (FDA approval, May 2019) is a multitarget anti-diabetic drug combination composed of metformin (MET), saxagliptin (SAX) and dapagliflozin (DAP). To our present knowledge, no analytical reports were found in the scientific databases for the simultaneous quantification of MET, SAX and DAP in their ternary combined tablets, moreover, no articles have attempted the simultaneous estimation of the cited drugs in any matrix using high-performance liquid chromatography with diode-array detection (HPLC–DAD) or high-performance thin-layer chromatography (HPTLC) technique. The current work represents a comparative study on two developed and validated chromatographic methods for the simultaneous determination of the ternary mixture (MET, SAX and DAP) in pure form and in combined tablet dosage form. The first method is reversed-phase HPLC using Agilent C18 column (4.6 × 250 mm, 5 μm p.s.) with a mobile phase consisting of acetonitrile and acidic aqueous phase pH 3 with a photodiode array detection at 230 nm. The second method is HPTLC in which drug solutions were applied to Merck HPTLC silica gel plates developed with a mixture of chloroform:methanol:water:acetic acid (7.4:2.6:0.5:0.01, v/v) and scanned at 224 nm. Both methods were fully validated following the ICH guidelines in terms of linearity, accuracy, precision, selectivity and robustness.
Representative HPLC (a) and HPTLC (b) chromatograms for a ternary mixture of metformin (MET), saxagliptin (SAX) and dapagliflozin (DAP)
Medium-pressure preparative liquid chromatography (MPPLC) was used to isolate and prepare lactucin and lactucopicrin from the whole herb of Cichorium glandulosum. After extracting the methanolic extract of the whole plant with petroleum ether and ethyl acetate several times to obtain ethyl acetate extract, the crude products, namely, lactucin and lactucopicrin were separated using MPPLC and thin-layer chromatography (TLC) tracking, and their purity rates reached more than 80%. The qualitative and quantitative analyses of lactucin and lactucopicrin were carried out by high-performance thin-layer chromatography (HPTLC) on the whole herb of C. glandulosum. The contents of lactucin and lactucopicrin were determined by scanning at 256 nm in the whole herb of C. glandulosum. The RF values of lactucin and lactucopicrin were 0.42 ± 0.05 and 0.65 ± 0.05 with linear ranges of 0.498–2.988 and 0.499–2.994 μg/band, respectively. The correlation coefficients were 0.9938 and 0.9946, respectively, thereby showing a good linear relationship. The average recoveries were 99.96% and 99.52%, and the relative standard deviations (RSDs) were 2.49% and 2.45%, respectively. The crude products, namely, lactucin and lactucopicrin, can be isolated with high purity from the whole herbs of C. glandulosum by MPPLC. The lactucin and lactucopicrin contents of C. glandulosum can be determined rapidly and accurately by HPTLC.
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 reverse-phase method for the selective quantification of pitavastatin calcium (PIT) and its related substances was developed. The method demonstrated an excellent separation between PIT and each of 15 impurities (including its isomers and degradants) within a short run time of 12 min by HPLC. A rapid resolution similar to that of UHPLC was achieved using high flow rate on superficially porous C18 stationary phase. A synergistic combination of quality by design approach and use of a superficially porous column delivered a HPLC method with ultra-high performance. Forced degradation studies proved the method to be highly specific (mass balance > 98 %) and the structures of major degradation products were proposed based on LC–MS analysis. The results of validation proved the method to be highly precise (%RSD < 4), accurate (recoveries in range of 100 ± 7 %), linear (r 2 > 0.999) and sensitive (LOQ ≤ 0.02 % and LOD ≤ 0.005 %) for all the impurities and drug. Use of multivariate analysis helped to incorporate high robustness in the method. The method is valuable for quantification of PIT and its related substances in both drug substance and oral solid dosage form.
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