A thin-layer chromatographic method has been established for quantification of aloenin in aloe (
Mill.) pharmaceuticals. Chromatographic separation was performed on silica gel plates with ethyl acetate-ethanol (95%)-water, 20:3:1 (
), as mobile phase. The plates were scanned densitometrically at 365 nm. The method was validated for precision, repeatability, and accuracy. It was found to be precise — intra-day and inter-day
were 2.21% and 3.15%, respectively. Instrumental precision and repeatability for the method were 0.42 and 1.94 (
[%]), respectively. Accuracy was checked by measurement of recovery at three levels; average recovery was 97.86%. The method was used for analysis of aloenin in aloe juice, aloe tablets and aloe liquid extract for injections and was confirmed to be suitable for this purpose.
A thin-layer chromatographic method has been established for quantification of kurarinone and sophoraflavanone G in the roots of Sophora flavescens Soland. (Fabaceae). Chromatographic separation was performed on silica gel HPTLC plates with chloroform-methanol 10:1 (ν/ν) as mobile phase. The plates were scanned densitometrically at 285 nm. The method was validated for precision, repeatability, and accuracy. It was found to be precise — intra-day and inter-day RSD were 1.97–2.04% and 2.15–2.40%, respectively. Instrument precision and repeatability of the method were 0.43–0.56 and 1.81–1.84 (CV [%]), respectively. Accuracy was checked by measuring recovery at four levels; average recovery was 96.3–103.4% for kurarinone and 96.8–102.9% for sophoraflavanone G. The method was used for analysis of kurarinone and sophoraflavanone G in Sophora flavescens roots and was confirmed to be suitable for this purpose.
Authors:Pritam Jain, Rahul Khatal, Hardik Jivani and Sanjay Surana
Asimple, selective, precise, and stability-indicating HPTLC method for analysis of brimonidine tartrate as the bulk drug and in formulations has been established and validated. Chromatography on aluminum foil silica gel 60F254 plates with methanol-toluene-triethylamine 1:3.5:0.2 (ν/ν) as mobile phase gave a compact band at RF 0.48 ± 0.02. Densitometric analysis was performed in absorbance mode at 247 nm. Linear regression analysis of peak-area calibration data revealed a good linear relationship with r2 = 0.9965 ± 0.0013 in the concentration range 100–600 ng per band. The mean value ± SD of slope and intercept were 9.047 ± 0.11 and 553.0 ± 39.06. The method was validated for precision, recovery, and robustness. The limits of detection and quantitation were 9.40 and 28.51 ng per band, respectively. Brimonidine tartrate was subjected to hydrolysis (in acid, alkali, and neutral solutions), oxidation, and photodegradation, and was degraded under all these conditions. Statistical analysis proved the method enables selective, precise, and accurate identification and quantitative analysis of brimonidine tartrate as the bulk drug and in dosage forms.
Authors:Daniil Olennikov and Vyacheslav Partilkhaev
Athin-layer chromatographic (TLC) method was used for the quantification of four dominant flavonoids (rutin, narcissin, nicotiflorin, isoquercitrin) in the shoots of Cargana spinosa (L.) DC. (Fabaceae). Chromatographic separation was performed on silica gel HPTLC plates with double development of ethylacetate-1,2-dichloroethane- acetic acid-85% formic acid-water 10:2.5:1:1:0.8 (v/v) as mobile phase. The plates were scanned densitometrically at 360 and 387 nm. The method was validated for precision, repeatability, and accuracy. It was found to be precise: intraday and interday RSD were 1.48–1.87% and 1.59–1.97%, respectively. Instrumental precision and repeatability for the method were found to be 0.51–0.64 and 1.18–1.32 (CV (%)), respectively. Accuracy was checked by measuring the recovery at four levels; the average recovery was 98.19–101.36% for rutin, 98.04–101.14% for narcissin, 98.16–100.54% for nicotiflorin, and 98.11–101.98% for isoquercitrin. The method was used for the analysis of the flavonoids mentioned in C. spinosa shoots samples. Nicotiflorin was detected in Caragana genus for the first time.
Authors:Hayriye Kurtbay, Ipek Kaynak, Serap Bozkurt and Melek Merdivan
A thin-layer chromatographic method with densitometric UV detection at 286 nm after ethyl acetate extraction is described for analysis of 5-hydroxymethylfurfural (HMF) in fruit wines and vinegars produced in Turkey. The retention factor,
, was 38 on silica gel HPTLC plates as stationary phase after development with toluene-ethyl acetate-90% formic acid 6:3:1 (
) as mobile phase. Recovery from samples spiked with different amounts HMF (2.0 and 5.0 ng μL
) was usually in the range 95–112%. The limits of detection and quantification of the method were 0.045 and 0.125 ng mL
, respectively. For robustness, within and between-day repeatability of the method were calculated as 4.5 and 8.6%, respectively. Seven types of Turkish fruit wines and three kind of Turkish vinegar tested were found to contain HMF in the ranges 0.5 to 13.2 ng μL
and 1.0 to 33.1 ng μL
Authors:Danuta Raj, Adam Kokotkiewicz and Maria Łuczkiewicz
An optimized chromatographic method has been established for separation of alkaloid fractions in the herb
and in different types of in-vitro cultures. TLC separation of
indolizidine alkaloids was optimized by using four types of plate, both silica gel (Si60 F
TLC plates, and Si60 F
and LiChrospher Si60 F
HPTLC plates) and reversed-phase (RP 18W), with a range of solvent mixtures as mobile phases. The best resolution was achieved with HPTLC Merck LiChrospher Si60 F
and chloroform-methanol 100:5 (
) as mobile phase. The method, with densitometric detection, was then used for quantitative analysis of the securinine and allosecurinine content of extracts from different
samples. The method was validated for linearity, limits of detection and quantification, repeatability, intermediate precision, and recovery.
A densitometric HPTLC method for analysis of cordifolioside A both in 60% methanolic extract of Tinospora cordifolia and in a commercial formulation has been established and validated. Cordifolioside A was separated on aluminum-backed silica gel 60 F254 plates with chloroform-methanol 85:15 (%, v/v) as mobile phase. A compact band was obtained for cordifolioside A at RF 0.52 ± 0.03. The limits of detection (LOD) and quantification (LOQ) were 20.12 and 60.36 ng per band, respectively. The highly precise and accurate method was used for analysis of cordifolioside A.
Asensitive, selective, and precise high-performance thin-layer chromatography (HPTLC) method was developed for simultaneous analysis of six bioactive phenolic compounds, i.e., juglone, quercetin, myricetin, rutin, caffeic acid, and gallic acid in methanol extract and its fractions (hexane, chloroform, ethyl acetate, and butanol fractions) from bark of Juglans regia. Good separation was achieved on RP-18 F254S TLC plate using methanol-water-formic acid-acetic acid (48.8:46.4:2.4:2.4, ν/ν). The densitometric determination of the compounds was carried out at 254 nm in reflectance/absorbance mode. The method was validated in terms of linearity, sensitivity, accuracy, precision, robustness, and specificity. The linear regression data for the calibration plots of the reference compounds showed a good linear relationship with higher correlation coefficient (r2 ≥ 0.997). Accuracy of the method was evaluated in terms of average percent recovery, which ranged from 98.63 to 101.06%. HPTLC results revealed qualitative and quantitative differences in the phenolic compounds in the extract and fractions. The ethyl acetate fraction contained gallic acid followed by myricetin, rutin, quercetin, and caffeic acid in higher amount in comparison to the extract and fractions. The present method can be used for routine quality control of J. regia extracts.
Linn, family Rubiaceae, is an important component of the ayurvedic system of medicine. Purpurin is the major anthraquinone present in
. It has an antigenotoxic activity. A sensitive, selective, precise, and robust high-performance thin-layer chromatographic (HPTLC) method of analysis of purpurin from the parts of
and pharmaceutical dosage forms has been developed and validated. The method employs aluminum foil HPTLC plates coated with silica gel 60F
as the stationary phase. The mobile phase was toluene-ethyl acetate-formic acid 9.8:0.2:0.1 (
). Densitometric analysis of purpurin was carried out in the absorbance mode at 255 nm. The method was validated for linearity (100–600 ng per band), precision (intra-day variation 0.28 to 1.39%, inter-day variation 0.72 to 2.21%), recovery (96.54 to 98.53%), robustness, and stability of purpurin. The limits of detection and quantification for purpurin were 50 and 100 ng per band, respectively. The method can be used for identification and quantification of purpurin in herbal extracts of
and in pharmaceutical dosage forms.
Authors:Katarzyna Szewczyk, Łukasz Komsta and Agnieszka Skalska-Kamińska
High performance thin-layer chromatography (HPTLC) combined with densitometry has been used for analysis of the triterpenoid content of acetone and ethyl acetate extracts of the leaves of
(Sims.) Opiz. After optimization of the extraction process, HPTLC separation was successfully standardized, using silica gel plates washed with methanol and dichloromethane, dichloromethane-ethyl acetate 18.5:1.5 as mobile phase, and 8% H
in ethanol (
) as spray reagent. Triterpenoids in the leaves of
were quantified by measurement of absorbance. Three spots were observed on chromatograms — the sum of α and β-amyrin, oleanolic acid, and an unidentified compound. All of the triterpenoid components were analyzed for the first time in this plant. The triterpenoid content was calculated statistically.