Gradient thin-layer chromatography and densitometry have been used for qualitative and quantitative analysis of caffeic acid in some Dipsacaceae family plants. The presence of caffeic acid was determined in complex plant extracts before and after acid hydrolysis.
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 (r 2 ≥ 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.
A thin-layer chromatographic (TLC)-densitometric method was used for the parallel quantification of rosmarinic acid (RA) and caffeic acid (CA) in crude extracts of Salvia species (Family Lamiaceae), obtained by ultrasonic extraction with 60% methanol. The densitometric measurement was performed in fluorescent mode as it has been published earlier. The applicability of the method has been investigated mainly from the viewpoint of the starting material. Questions are discussed like, what kind of factors should be taken into account, if the drugs are to be characterized, and how the RA and CA contents of samples vary in the plants are discussed. The drugs (plant material) show great differences due to the time of harvest, to the organ composition of drugs, and to the extraction and storage conditions of the stock-solutions prepared from them. The importance of these parameters is illustrated on Salvia species native to Hungary.
Secondary metabolites are an integral part of the plant kingdom which plays a vital role in development, growth, and defense. These are also very helpful for humans against different disorders. Phenols due to their strong antioxidant activity have a central role in conferring different types of biological activities. The present study aims at the simultaneous determination of chlorogenic acid and caffeic acid in the methanolic extracts of leaf samples of Siegesbeckia orientalis L. Simultaneous elution of both compounds has been attained by using toluene‒ethyl acetate‒formic acid‒methanol (6:6:1.6:0.6 v/v) as the mobile phase, and densitometric evaluation was done at 366 nm. The linear regression data for the calibration curve of both compounds show good linear relationship in the concentration range of 200–600 ng. The proposed method has been validated in terms of limit of detection, limit of quantification, specificity, precision, and accuracy studies. The present high-performance thin-layer chromatographic method has been applied for the quantification of both compounds in the various accessions of S. orientalis collected from different localities of North India. Maximum amount of chlorogenic acid was found in the Taluka accession, while caffeic acid was found in the Rarhi accession collected from Uttarakhand state.
The present paper reports a validated high-performance thin-layer chromatography (HPTLC)‒densitometric method for the simultaneous quantification of phenolic (ferulic acid and caffeic acid) and terpenoid (β-sitosterol and lupeol) markers in Convolvulus pluricaulis Choisy. According to Ayurveda, it is commonly known as ‘Shankhpushpi’ due to its ‘Conch’ or ‘Shankh’-shape flower. The plant species, viz., Clitoria ternatea L., Evolvulus alsinoides (L.) L., and Tephrosia purpurea (L.) Pers., also having similar flowers are reported as its adulterants/substitutes. This creates a problem in its quality and efficacy in the commercial drug market of India. Therefore, a HPTLCmethod was performed on a pre-coated silica gel 60 F254 plate with the aforesaid markers. The solvent system toluene–ethyl acetate–formic acid (8.5:1.5:0.1) was determined to be the best system for the simultaneous separation of caffeic acid, ferulic acid, β-sitosterol, and lupeol at R F values of 0.14, 0.29, 0.48, and 0.63, respectively. A densitometric scanning profile of all the samples at 580 nm showed peaks for all the four markers of varying heights in the samples, except the absence of caffeic acid in Tephrosia purpurea. The developed method was standardized and validated for the quantification of active principal-based quality-control markers in terms of precision, accuracy, linearity, recovery, and repeatability. It will help to maintain batch-to-batch consistency and identification of adulterants/substitutes in raw materials during production of drug in the pharmaceutical units.
Orthosiphon stamineus Benth., known as Java tea, is an important medicinal plant used for its diuretic action. Orthosiphon stamineus Benth. leaves contain many active compounds; some of the most important are the caffeic acid derivatives. Extraction of caffeic acid derivatives was optimized by use of the ‘Prisma’ strategy. Extracts were evaluated by spectrophotometry and TLC. Total polyphenol and rosmarinic acid content were determined qualitatively and quantitatively. It was observed that a ternary mixture of methanol, water and methyl acetate, 1 + 1 + 8 ( v/v ), extracted more caffeic acid derivatives than when the solvents were used individually.
Oxalis corniculata is an important medicinal plant, but chromatographic analysis for the simultaneous quantification of different phenolic compounds has not been performed till date in this plant species. A high-performance thin-layer chromatography (HPTLC) method for the simultaneous quantitative determination of caffeic acid, vanillic acid, and syringic acid in O. corniculata L. methanolic fraction was developed for the first time. In HPTLC, for achieving good separation, the mobile phase of toluene‒ethyl acetate‒formic acid (7:3:1, v/v) was used. The densitometric determination was carried out at 300 nm in reflection/absorption mode. The calibration curves were linear in the range of 100–700 ng per spot for caffeic acid, vanillic acid, and syringic acid. HPTLC analysis has indicated the presence of optimum amount of caffeic acid (0.025 ± 0.0002), vanillic acid (0.018 ± 0.0005), and syringic acid (0.04 ± 0.0006) in the methanolic fractions. The optimized method was successfully applied for the analysis of three major phenolics in O. corniculata L. The proposed method is simple, precise, specific, and accurate. The quantification of these phenolic compounds has not yet been reported in this species which may be utilized for the proper standardization of the drug.
A high-performance thin-layer chromatographic (HPTLC) method for the simultaneous quantitative determination of caffeic acid, vanillic acid, syringic acid and kaempferol in flowers and buds of three different Bauhinia species was developed for the first time in the case of these species. Methanol was found to be the best for the highest possible recovery of 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 350 nm in reflection—absorption mode. The calibration curves were linear in the range of 100–700 ng per spot for caffeic acid, vanillic acid, syringic acid, and kaempferol. The methanolic fractions of Bauhinia variegata L. flowers (BVFM) showed the highest amount of caffeic acid (0.08%), B. variegata L. buds (BVBM) and Bauhinia purpurea L. flowers (BPFM) showed the highest amount of kaempferol (1.53%), Bauhinia acuminata L. flowers showed the highest amount of vanillic acid (0.40%), and B. acuminata L. buds showed the highest amount of syringic acid (0.08%). The proposed method is simple, precise, specific, accurate, less time-consuming, and cost-effective. The statistical analysis of data obtained proves that the method is reproducible and selective and can be used for routine analysis of reported phenolic compounds in crude drug and extracts. The simultaneous quantification of these phenolic compounds has not yet been reported in the case of these species which may be utilized for the proper standardization of the drug.
The healthy properties of functional food are associated with the presence of a wide range of phenolic antioxidants. Thus, the aim of this study was to develop a simple, rapid, and reproducible high-performance thin-layer chromatographic (HPTLC) method combined with a direct 2,2′-diphenyl-1-picrylhydrazyl (DPPH) assay, to compare antioxidant activity in terms of free radical scavenging activity in functional food. We wanted to quantify caffeic acid, chlorogenic, and gallic acid, three natural phenolic antioxidants commonly found in functional food, and determine their contribution to the total free radical scavenging activity. Antioxidant activities of organic cacao, green coffee, chia seeds, goji berries, spirulina, and chlorella were evaluated and compared. The most potent free radical scavenger was found to be chlorogenic acid. Free radical scavenging activity in the functional foods tested was mostly affected by the presence of chlorogenic and caffeic acid, while gallic acid, although present in higher amounts, was a less potent free radical scavenger.
absorbance of various phenolic substances at 10 min, it was necessary to prepared solutions: 0.0676 mM gallic acid; 0.0969 mM syringic acid; 0.1071 mM caffeic acid; 0.0910 mM sinapinic acid; 0.7361 mM vanillin, and 0.4994 mM Trolox. For comparison purposes