A validated high-performance thin-layer chromatography method for the determination of two bioactive lignans,phyllanthin and hypophyllanthin,in the seasonal variation study of Phyllanthus amarus

JPC – Journal of Planar Chromatography – Modern TLC - Phyllanthus amarus Schum. & Thonn. belongs to the family Euphorbiaceae, commonly known as “Bhooamalakee” or...     

A rapid densitometric method for simultaneous quantification of gallic acid and ellagic acid in herbal raw materials using HPTLC

Gallic acid and ellagic acid are two widely occurring phenolic compounds of plant origin, to which many biological activities including anticancer and antiviral activity have been attributed. A simple HPTLC method has been developed for the simultaneous quantification of gallic acid and ellagic acid. The method was validated for precision, repeatability, and accuracy. Instrumental precision was found to be 0.083 and 0.78, and the repeatability of the method was found to be 1.07 and 1.50 (% CV) for gallic acid and ellagic acid, respectively. The accuracy of the method was checked by a recovery study conducted at two different levels and the average percentage recovery was found to be 101.02% for gallic acid and 102.42% for ellagic acid. The above method was used for the quantification of gallic acid and ellagic acid content in seeds of Abrus precatorius Linn., whole plant of Phyllanthus maderaspatensis Linn., and flowers of Nymphaea alba Linn. The proposed HPTLC method for the simultaneous quantification of gallic acid and ellagic acid was found to be simple, precise, specific, sensitive, and accurate and can be used for routine quality control of herbal raw materials and for the quantification of these compounds in plant materials.     

A validated quantification of gallic acid and ellagic acid in Triphala using a high-performance thin-layer chromatography method

JPC – Journal of Planar Chromatography – Modern TLC - A well-known Ayurvedic preparation, Triphala, consists of Amla (Emblica officinalis Gaertn.), Baheda (Terminalia bellirica Roxb.)...     

Simultaneous identification of rutin,chlorogenic acid and gallic acid in Moringa oleifera by densitometric high-performance thin-layer chromatography method

JPC – Journal of Planar Chromatography – Modern TLC - An HPTLC (high-performance thin-layer chromatography) method was carried out to develop a feasible, simple and accurate method for...     

Validated high-performance thin-layer chromatographic method for quantification of gallic acid and ellagic acid in fruits of Terminalia chebula,Phyllanthus emblica,and Quercus infectoria

A simple and rapid high-performance thin-layer chromatographic method for quantification of gallic acid and ellagic acid in dried fruits of Terminalia chebula, Phyllanthus emblica, and Quercus infectoria has been developed. The chromatographic development was carried out on precoated silica gel 60 F₂₅₄ plates in a mixture of toluene:ethyl acetate:chloroform:formic acid (4:8:1:3 v/v/v/v). The plate was scanned densitometrically at a wavelength of 280 nm. The retention factor value of gallic acid and ellagic acid was found to be 0.63 ± 0.2 and 0.53 ± 0.1, respectively. The developed method was validated in terms of linearity, precision, accuracy, sensitivity, robustness, specificity and stability as per the international conference of harmonization guidelines. The method showed good linear relationship over a range of 100–600 ng/band (gallic acid) and 100–500 ng/band (ellagic acid) with a regression coefficient (r²) of 0.997 (gallic acid) and 0.996 (ellagic acid). The method showed high accuracy (99.65%–100.85%). The percentage relative standard deviation of intra-day and inter-day precision studies was not more than 2%. The method is highly robust and has displayed high specificity. The developed method is new, simple, and accurate and can be successfully employed in routine analysis of raw materials and formulations containing gallic acid and ellagic acid.     

High-performance liquid chromatography and thin-layer chromatography for the simultaneous quantitation of rabeprazole and mosapride in pharmaceutical products

Simple, sensitive high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC) methods are developed for the quantitative estimation of rabeprazole and mosapride in their combined pharmaceutical dosage forms. In HPLC, rabeprazole and mosapride are chromatographed using 0.01M 6.5 pH ammonium acetate buffer-methanol-acetonitrile (40:20:40, v/v, pH 5.70+/-0.02) as the mobile phase at a flow rate of 1.0 mL/min. In TLC, the mobile phase is ethyl acetate-methanol-benzene (2:0.5:2.5, v/v). Both the drugs are scanned at 276 nm. The retention times of rabeprazole and mosapride are found to be 4.93+/-0.01 and 9.79+/-0.02, respectively. The Rf values of rabeprazole and mosapride are found to be 0.42+/-0.02 and 0.61+/-0.02, respectively. The linearities of rabeprazole and mosapride are in the range of 400-2000 ng/mL and 300-1500 ng/mL, respectively, for HPLC; in TLC, the linearities of rabeprazole and mosapride are in the range of 400-1200 ng/spot and 300-900 ng/spot, respectively. The limit of detection is found to be 97.7 ng/mL for rabeprazole and 97.6 ng/mL for mosapride in HPLC; in TLC the limit of detection is found to be 132.29 ng/spot for rabeprazole and 98.25 ng/spot for mosapride. The proposed methods can be applied to the determination of rabeprazole and mosapride in combined pharmaceutical products.     

High-performance thin-layer chromatography densitometric method for the quantification of harmine, harmaline, vasicine, and vasicinone in Peganum harmala

Peganum harmala Linn. seed is a reputed drug of the Indian systems of medicine. We report a simple high-performance thin-layer chromatography (HPTLC) densitometric method for the quantification of 4 alkaloids, viz., harmine, harmaline, vasicine, and vasicinone from P. harmala seed. The 4 compounds were resolved in ethyl acetate-methanol-ammonia (7 + 1 + 0.3, v/v/v) mobile phase. The method was validated for precision, repeatability, and accuracy. Instrumental precision was 0.27, 1.53, 0.39, and 1.15% [relative standard deviation (RSD)] and repeatability of the method was 1.01, 0.79, 0.13, and 1.62% RSD for harmine, harmaline, vasicine, and vasicinone, respectively. Accuracy of the method was checked by a recovery study conducted at 3 different levels, and the average recovery was 97.88% for harmine, 97.69% for harmaline, 98.38% for vasicine, and 98.28% for vasicinone. The 4 compounds were quantified in P. harmala seed using the method, and it was found to contain 0.44, 0.096, 0.25, and 0.0007% (w/w) of each, respectively. The proposed HPTLC densitometric method for the quantification of the 4 compounds was found to be simple, precise, specific, sensitive, and accurate. It can be used for routine quality control of P. harmala seed and has the ability to quantify the 4 marker compounds in the formulations containing P. harmala. It also can be used to quantify any of these marker compounds in other herbal drugs.     

High-performance thin-layer chromatography densitometric method for the simultaneous determination of three phenolic acids in Syzygium aromaticum (L.) Merr. & Perry

A simple, precise, and rapid high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous determination of 3 phenolic acids, i.e., gallic acid, caffeic acid, and syringic acid, in the dried buds of Syzygium aromaticum, commonly known as clove. HPTLC was performed on silica gel 60F254 plates with toluene-ethyl acetate-formic acid (8 + 2 + 1) mobile phase and densitometric scanning at 280 nm. The method was validated for selectivity, linearity, precision, and repeatability. Instrumental precision coefficient of variation (CV) was 0.88, 0.93, and 0.98% and repeatability of the method (CV) was 0.76, 0.64, and 0.69% for gallic acid, caffeic acid, and syringic acid, respectively. The linear concentration ranges were 400-3200 ng/spot with a correlation coefficient of 0.993 for gallic acid, 440-3520 ng/spot with a correlation coefficient of 0.994 for caffeic acid, and 400-4000 ng/spot with a correlation coefficient of 0.993 for syringic acid. The average recoveries of gallic acid, caffeic acid, and syringic acid were 96.3, 95.7, and 92.4%, respectively. Gallic acid, caffeic acid, and syringic acid were present at levels of 1.58, 0.06, and 0.05% (w/w), respectively, in S. aromaticum. This method is simple, accurate, precise, and economical and can be used for routine quality control.     

Development and validation of a high-performance thin-layer chromatography densitometric method for the simultaneous determination of novel 1-acridinyl-1,2,3-triazole derivatives

JPC – Journal of Planar Chromatography – Modern TLC - High-performance thin-layer chromatographic (HPTLC) method provides a simple, sensitive, and accurate analytical method for the...     

A quantitative densitometric method for the rapid separation and quantitation of the major lipids of tissues and lipoproteins by high-performance thin-layer chromatography. II. Reduction of the densitometric data

A BASIC program is described for acquisition of data and data reduction for an automated densitometric system for quantitation of lipids separated by high-performance thin-layer chromatography. The program allows calculation of mass of samples from log/log calibration curves computed from standards. The calculated masses are reported as nmol/volume or nmol/mg protein. The program contains a flexible dialog system which permits its use for a variety of applications in addition to the system described for quantitation of lipids.     

A quantitative densitometric method for the rapid separation and quantitation of the major tissue and lipoprotein lipids by high-performance thin-layer chromatography. I. Sample preparation, chromatography, and densitometry

A rapid method for the separation and quantitation of the major lipids of tissues and lipoproteins by automated high-performance thin-layer chromatography is presented. Solvent systems for one-dimensional separation of neutral lipids, of cholesteryl esters, and of phospholipids are described. Separated lipids are measured following treatment with methanolic sulphuric acid containing manganese chloride and scanned in fluorescence or absorption mode. Absolute quantitation is obtained by the use of an internal standard and by references to standards for each lipid run on the same plates as samples. The method described here is particularly suitable for the rapid quantitation of small amounts of lipid (0.01-0.02 nmol per sample), for example in tissue culture studies; 100 micrograms of fibroblast or macrophage protein are sufficient for complete lipid analysis. The coefficients of variation due to the sample preparation, application to the plates and densitometry are in the range 7.2-9.1%. The method was compared with enzymatic determinations for cholesterol and gave correlation coefficients of 0.95 for total cholesterol and 0.91 for unesterified cholesterol. Phospholipid estimation was compared with large-plate thin-layer chromatography and phosphorus analysis and gave correlation coefficients of 0.90 for phosphatidylcholine and 0.89 for sphingomyelin.     

A new method for the analysis of seized Ya-Ba tablets by densitometric thin-layer chromatography

JPC – Journal of Planar Chromatography – Modern TLC -     

High-performance liquid chromatography method for simultaneous determination of aliphatic acid, aromatic acid and neutral degradation products in biomass pretreatment hydrolysates

A variety of degradation products are produced upon dilute acid pretreatment of lignocellulosic biomass. Within this larger construct, organic acids, phenols and aromatic aldehydes represent important compound classes to investigate due to increasing evidence of their inhibitory effect on fermentative microorganisms. An analytical extraction procedure is presented, enabling isolation of potential analytes away from alternative products in biomass hydrolysates. Additionally, a reversed-phase high-performance liquid chromatography (HPLC) method has been developed and validated, affording simultaneous separation and quantitative determination of 32 potential analytes in water with UV detection at 210 nm. The method was subsequently employed to quantify a variety of aliphatic acid, aromatic acid, aldehyde and phenolic degradation products in a corn-stover hydrolysate at concentration levels ranging from 0.02 to 41 mM.     

High-performance thin-layer chromatography method for monitoring norfloxacin residues on pharmaceutical equipment surfaces

This paper presents a high-performance thin-layer chromatography (HPTLC) method with direct fluorescence measurement for the determination of norfloxacin. The method was validated for the monitoring of norfloxacin residues on stainless steel surfaces at the allowed limit of 10 mg of norfloxacin per square meter. However, it can be adapted for lower amounts of residues owing to the low detection limit of norfloxacin (about 5 ng) and can also be used for other surface materials. Test solutions were analyzed by the new HPTLC method and the known HPLC method for comparison. Accuracy and precision of the new HPTLC method, with a subsequent quantification by densitometer or video system, are comparable with those of the HPLC method.     

Simultaneous determination of bergenin and gallic acid in Bergenia ligulata wall by high-performance thin-layer chromatography

A simple and reproducible high-performance thin-layer chromatographic method was developed for the simultaneous determination of bergenin and gallic acid in Bergenia ligulata. Water and methanol were used as the extracting solvents. The concentrations of bergenin and gallic acid in both of these solvents were found to be almost the same. The method involves separation of the components by thin-layer chromatography on a precoated Silica Gel 60 F254 plate with a solvent system of ethyl acetate-formic acid-acetic acid-water (100 + 11 + 11 + 27). The sensitivity of the method for bergenin was 0.30 microg, whereas for gallic acid it was 0.25 microg. The proposed method is precise and sensitive and can be used for the detection, monitoring, and simultaneous quantification of bergenin and gallic acid in B. ligulata.     

A validated method for the thin-layer chromatographic in situ autofluorescence densitometric quantitation of the benzylisoquinoline alkaloids berberine and sanguinarine

JPC – Journal of Planar Chromatography – Modern TLC - A thin-layer chromatography method for the separation and in situ densitometric quantitation of sanguinarine and berberine was...     

Development and validation of a high-performance thin-layer chromatography method for the simultaneous quantification of rosmarinic acid,quercetin, glycyrrhizin and betulinic acid in polyherbal immunostimulant formulation

JPC – Journal of Planar Chromatography – Modern TLC - Quantification of bioactive markers through modern analytical methods is very essential for establishing the authenticity and...