Authors:Urszula Hachuła, Sabina Anikiel, and Marzena Połowniak
This paper describes rapid and cost-effective analytical procedures for determination of gallic acid after chromatographic separation. Optimum conditions were established for separation and assay of gallic acid, salicylic acid, and tannic acid by thin-layer and paper chromatography (PC). Detection was by means of a sensitive spectrophotometric method based on the coupled redox-complexation reactions occurring in the system Fe(III), gallic acid, and 2,2′-dipyridyl (
= 522 nm). The procedure was then applied to the determination of gallic acid in a pharmaceutical preparation. Thin layer chromatographic analysis with densitometric UV detection at λ = 280 nm gave comparable results.
A simple, highly precise method has been established for the simultaneous determination of the bioactive molecules bergenin and gallic acid in three different species of
(Royle) Raizada, and
Engl. The assay combines separation and quantitative estimation of the analytes on silica gel 60GF
HPTLC plates with visualization under UV light and scanning at 260 nm. This study has enabled simultaneous analysis of bergenin and gallic acid in all three species.
Authors:Renata Slaveska-Raicki, Vesna Rafajlovska, Vesna Rizova, and Ilinka Spirevska
A quantitative high-performance thin-layer chromatographic (HPTLC) method for analysis of gallic acid and tannin in extracts of
(L.) Sprengel, bearberry leaves (Ericaceae), and validation of the method, are described. Analyses were performed on cellulose F
HPTLC plates with
-butanol-acetic acid-water, 14 + 1 + 3.5 (
), as mobile phase. The chromatographic system afforded satisfactory
(0.35 for gallotannin and 0.46 for gallic acid), low
(0.1–0.44%), and effective resolution between components (
= 1.89). The
leaves examined contained approximately 14% tannin and 5% gallic acid. Special features of the thin-layer chromatographic (TLC) technique were taken into consideration in the validation process and calculation of validation data for the method. This HPTLC method was found to be simple, reliable, and convenient for routine analysis. Its analytical performance fulfilled acceptance criteria established for TLC methods in the official literature.
Authors:Urszula Hachuła, Sabina Anikiel, and Mieczysław Sajewicz
Tea is one of the most popular beverages in the world. The exact composition of tea depends on the manufacturing process. Our goal was to optimize the chromatographic conditions for separation of tea extract and to apply the optimized method for determination of gallic acid in oolong, black, and pu-erh tea. Chromatographic separation was performed on silica gel 60 F
TLC plates with chloroform-ethyl acetate-formic acid, 5 + 4 + 1 (
), as mobile phase. Densitometric measurement was performed in the ultraviolet region at
= 280 nm. Quantification of gallic acid in pu-erh tea was also performed by a spectrophotometric method based on the coupled redox complexation reaction occurring in the system Fe(III)-gallic acid-2,2′-dipyridyl (
= 522 nm).
Authors:Sadia Shakeel, Somia Gul, Aqib Zahoor, Saleha Suleman Khan, Zeeshan Ahmed Sheikh, Safila Naveed, and Khan Usmanghani
The technological improvement in the structural elucidation of natural compounds has made it probable to generate appropriate strategies for the analysis and standardization of plant-based medicines. An appliance of highly oriented hyphenated techniques provides a definite tool for herbal investigations. Therefore, the present study was directed towards the standardization of biomarkers gallic acid and berberine in polyherbal formulation Entoban capsules to ensure the quality of the herbal drugs. A rapid, simple, accurate, and specific high-performance thin-layer chromatography (HPTLC) method for the quantitative estimation of biomarkers berberine and gallic acid has been developed. HPTLC was performed to evaluate the presence of gallic acid and berberine applying toulene—ethyl acetate—formic acid—methanol (12:9:4:0.5 v/v) and ethanol—water—formic acid (90:9:1 v/v), as the mobile phase, respectively. The RF values (0.58 for gallic acid and 0.76 for berberine) in both sample and reference standard were found comparable under ultraviolet (UV) light at 273 nm and 366 nm, respectively. The method developed resulted in good-quality peak shape and enabled high-quality resolution of biomarkers. The present standardization undertaken reveals compliance with the analytical procedure; therefore, it is concluded that Entoban capsule is a well-standardized product. Standardization falls under the specific guidelines of quality herbal medicine following the prerequisite for global harmonization.
Authors:Sachin Navale, Vinod Kuber, and Shrinivas Bhope
A densitometric HPTLC method has been established for simultaneous quantification of sennosides A and B and gallic acid in a laxative polyherbal pharmaceutical dosage form. Aluminum plates coated with silica gel 60 F254 as stationary phase were used with toluene-ethyl acetate-formic acid-methanol 8:8:4:5 (v/v) as mobile phase. Densitometric analysis was performed at 270 nm. Amounts of sennosides A and B and gallic acid were 10.48, 9.03, and 2.96% (w/w) respectively. The method was validated for specificity, linearity, precision, repeatability, and accuracy. Calibration plots were linear in the concentration range 114.0–427.5 ng per band for sennosides A and B and 100–375 ng per band for gallic acid. The correlation coefficients were 0.995, 0.998, and 0.997 for sennosides A and B and gallic acid, respectively. Relative standard deviation (RSD, [%]) for instrumental precision and repeatability of the method was 0.74, 0.49, 0.43 and 1.11, 1.08, 0.89 for sennosides A and B and gallic acid, respectively. Recovery was 96.28–97.22% for sennoside A, 98.06–100.84% for sennoside B, and 97.09–98.07% for gallic acid. The method is simple, precise, specific, accurate, and economical. It can be used for routine analysis of formulations containing Senna (Cassia angustifolia) and Haritaki (Terminalia chebula) extracts.
Authors:Subrata De, Pankaj Nariya, and Nikhil Jirankalgikar
Piperine and gallic acid are of different chemical natures — piperine is an alkaloid, while gallic acid a phenolic compound. They are used as marker compounds in many plant-based formulations. A highperformance thin-layer chromatographic (HPTLC)-densitometric method has been developed and validated for the simultaneous quantification of piperine and gallic acid as such and in pharmaceutical dosage forms. Toluene-ethyl acetate (3:7) was used as mobile phase and scanning was done at 254 and 340 nm. The method was validated with respect to linearity, reproducibility, specificity, accuracy, precision, robustness, and ruggedness. Both compounds showed good linearities in the range of 250–1750 ng. LOD and LOQ for piperine were 9.98 and 33.29 ng, while for gallic acid 25 and 83.33 ng. Average % RSD values of precision for piperine and gallic acid were 0.46% and 0.72%, respectively. % Recovery was 96–103%. The method is accurate, reproducible, cost-effective, and can be used in routine analysis.
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.
Authors:Ankita Misra, Sharad Srivastava, and Ajay Kumar Singh Rawat
A validated online over-pressured layer chromatography (OPLC) method was developed for the rapid, cost-effective, and efficient separation of gallic acid. A reversed-phase (RP) ultraviolet (UV)–online OPLC analytical method was developed and validated as per the International Conference on Harmonization (ICH) guidelines. Methanolic extract of Annona muricata (fruit) was prepared by cold maceration. Separation of marker was achieved on RP-OPLC plates (5 × 20 cm) with isocratic solvent system of 0.1% acetic acid, water and methanol (30:70) at a flow rate of 0.15 mL min−1; detection was carried out at λmax 270 nm. The base peak of standard gallic acid was at 5.441 min with good linearity (r2 > 0.999), precision, and accuracy. The limit of detection (LOD) (521.84 mg mL−1) and limit of quantification (LOQ) (1581.336 mg mL−1) values reflect that the method is sensitive with high peak purity; the recoveries of analyte were 99 to 103%. The achievement of the method was the early retention time of gallic acid which in turn increased the efficiency of the quantification of the targeted marker in a short duration of time for even larger number of samples in plant extract as well as biological fluids for pharmacokinetic studies. The application of the method can be extended in regulatory guidelines for the quality control of herbal drugs/products and formulations. The method is rapid and economical in terms of solvent consumption and, hence, can be preferred over other high-performance thin-layer chromatographic or liquid chromatographic methods.
A high-performance thin-layer chromatography (HPTLC) method for the simultaneous quantitative determination of gallic acid, vanillic acid, protocatechuic acid, and quercetin in methanolic fractions of Limonia acidissima L. fruits was developed for the first time for this species. Methanol was found to be the best for the highest possible recovery of the target analytes. For achieving good separation, a mobile phase of toluene–ethyl acetate–formic acid (5:4:1 v/v) was used. The densitometric determination was carried out at 310 and 254 nm in reflection–absorption mode. The calibration curves were linear in the range of 100–600 ng per spot for gallic acid, vanillic acid, protocatechuic acid, and quercetin. The methanolic fractions of L. acidissima L. fruits showed the presence of gallic acid (0.07%), vanillic acid (0.16%), protocatechuic acid (0.06%), and quercetin (0.14%). The proposed method is simple, precise, specific, and accurate. The statistical analysis of the data obtained proves that the method is reproducible and selective and can be used for the routine analysis of the reported phenolic compounds in crude drug and extracts. The simultaneous quantification of these compounds has not been reported yet in L. acidissima L., which may be utilized for the proper standardization of the drug.