Simultaneous Determination Of Phenolic Acids Research Articles

A simple HPLC-DAD method for analysis of phenolic acids: Addition effect of a hydrophilic deep eutectic solvent to the mobile phase

AbstractThe analysis of phenolic acids (PAs) is of great importance, because they are frequently present in natural products and their derivatives, and these compounds also have multiple beneficial effects to human health. This work is focusing on the separation of seven PAs (caffeic acid, coumaric acid, gallic acid, ferulic acid, protocatechuic acid, sinapic acid, and syringic acid), in a reversed-phase liquid chromatographic (RP-HPLC) isocratic method using a hydrophilic deep eutectic solvent (DES) as a mobile phase additive. The analysis was carried out with a diode array detector. The used DES was composed by choline chloride and glycerol, and it was characterized by infrared spectroscopy. The combination of choline chloride:glycerol (1:4) added at 0.25% to mobile phase composed of 0.15% formic acid aqueous solution and methanol (80:20), showed the best separation for target analytes. The new proposed method was validated, and results indicated that the proposed method is linear, selective for almost all analytes, provided high sensitivity with limit of detection ranges from 0.009 to 0.023 mg mL−1, and has satisfactory precision and accuracy, with values of relative standard deviation of 0.24–2.65% and recoveries of 97.97–109%, respectively. Additionally, this method was successfully applied to simultaneous determination of phenolic acids in three kinds of samples of powder to prepare lemon flavour drink enriched with black tea extract.

Antioxidant, Anti-Obesity, and Hypolipidemic Effects of Polyphenol Rich Star Anise (Illicium verum) Tea in High-Fat-Sugar Diet-Induced Obesity Rat Model.

Star anise (Illicium verum Hook. fil.) is commonly utilized as a culinary and medicinal fruit and is most famous in indigenous systems of medicine. The present research work aims to appraise and validate the potential of polyphenol-rich star anise tea (SAT) on oxidative stress, obesity and related biochemical parameters in high-fat-sugar-diet (HFSD)-induced obesity model in rats. SAT was prepared using the traditional method in warm water. The Reverse Phase High Pressure Liquid Chromatography (RP-HPLC) analysis was performed for the simultaneous determination of phenolic acids and flavonoids in SAT. Two doses (250 and 500 mg/kg body weight) were selected to investigate the anti-obesity potential of SAT using HFSD-induced obese rat model. Major (>5 mg/100 mL) phenolic acids in SAT were p-coumeric acid, gallic aid, cinamic acid, chlorogenic acid and ferulic acid while catechin and rutin were the major flavonoids detected in the SAT. SAT exhibited 51.3% DPPH radical scavenging activity. In vivo study showed that higher doses of SAT (500 mg/kg body weight) significantly reduced the body weight increase (74.82%) and BMI (0.64 g/cm2). Moreover, significant reductions in the levels of serum total cholesterol, triglyceride, LDL and VLDL were recorded in all the treatment groups in comparison to the HFSDC group. Furthermore, SAT reduced the alterations in MDA, SOD and GSH levels of experimental groups thus showing the potential against oxidative stress. The SAT-500 group showed a significant decrease in the elevated kidney and liver weights and atherogenic index in comparison to the HFSDC group. The present study proved that SAT exhibited strong protective effects against obesity and oxidative stress, especially at higher doses.

Simultaneous Determination of Phenolic Acids, Anthraquinones, Flavonoids, and Triterpenes of Cynomorii Herba in Different Harvest Times by LC-MS/MS.

A sensitive and accurate method was developed for the simultaneous determination of twelve components including phenolic acids (gallic acid, protocatechuic aldehyde, protocatechuic acid, and ferulic acid), flavonoids (catechin, epicatechin, rutin, luteolin, luteolin-7-glucoside, and epicatechin gallate), anthraquinones (emodin), and triterpenes (ursolic acid) in Cynomorii herba in different harvest times by liquid chromatography tandem mass spectrometry (LC-MS/MS). The chromatographic separation was achieved on an Eclipse plus C18 (3.0 mm × 50 mm, 1.8 μm) column at 40°C. The mobile phase consisted of acetonitrile and 0.05% formic acid with a gradient elution at a flow rate of 0.4 mL·min −1. Under the optimized conditions, there was good linear relation (r ≥ 0.9944) and satisfactory precision (RSD values less than 5.65%). The recoveries of the twelve components were in the range of 93.5–105%. Moreover, the limits of detection (LOD) ranged from 0.003 to 21 ng mL−1 for the twelve analytes. In conclusion, the validated method was successfully applied to analyze the change regularity of the twelve components of Cynomorii herba in different harvest times. It provides a theoretical basis for choosing the suitable harvesting time of Cynomorii herba.

Simultaneous determination of phenolic acids and flavonoids in Artemisiae Argyi Folium by HPLC-MS/MS and discovery of antioxidant ingredients based on relevance analysis

A rapid and reliable LC–MS/MS method was developed for the quantitation of major components in Folium Artemisiae Argyi (mugwort), a widely used traditional Chinese herbal medicine. A total of 5 phenolic acids and 17 flavonoids were separated and simultaneously determined by using a Shiseido C18 column (150 × 3.0 mm, 3 μm) and gradient elution of acetonitrile-aqueous formic acid (100:0.1, v/v) at a 0.5 mL min−1 flow rate, via multiple reaction monitor (MRM) in polarity switching mode. The quantitative method was validated in terms of sensitivity, linearity, precision, accuracy and stability, which proved to be sensitive, accurate and reproducible. Then 65 samples collected from different areas were selected for component analysis by LC–MS/MS and assessment of antioxidant activity using DPPH, ABTS, FRAP, O2− and OH scavenging assays. Grey relational analysis and partial least square regression were used to evaluate the relevance between chemicals and bioactivities, and the results indicated chlorogenic acid, isochlorogenic acid B, A, C, eriodictyol, jaceosidin and eupatilin made the key contribution to antioxidant activity. The present study combines chemical analysis and bioassay to identify bioactive markers, which possesses potential value for the activity-oriented quality control of mugwort.

Simultaneous Determination of Phenolic Acids, Anthraquinones, and Flavonoids in the Aerial Part and the Fruit of Xanthium Sibiricum by LC- ESI- QTRAP-MS/MS

Background: Xanthium sibiricum is a well-known traditional Chinese medicine (TCM) that has been commonly used to treat rhinitis and related nasal diseases. The aim of this study was to develop a comprehensive analytical method based on high-performance liquid chromatographyelectrospray ionization coupled with triple quadrupole-linear ion trap mass spectrometry (LC-ESIQTRAP- MS/MS) for the simultaneous determination of phenolic acids, anthraquinones, and flavonoids in the aerial part and fruit of Xanthium sibiricum. Methods: The separation was completed on Agilent ZORBAX SB-C18 column (250 × 4.6 mm, 5μm) using methanol and 0.2% (v/v) aqueous formic acid as the mobile phase. The target components were analyzed in negative ion mode with accurate and sensitive multiple reaction monitoring (MRM) mode. Results: The correlation coefficients of all the calibration curves were higher than 0.9994. Relative standard deviations of intra- and inter-day precisions of the eighteen components were all lower than 2.87% and the recoveries were in the range from 97.73% to 101.82%. The validated method was successfully applied to possess forty Xanthium sibiricum samples (Xanthii Herba, Xanthii Fructus, and processed Xanthii Fructus) collected from different places in P. R. China. Furthermore, principal component analysis (PCA) was performed to evaluate and classify the samples according to the contents of the eighteen bioactive components. Conclusion: All the results demonstrated that the developed method was useful and could be applied for the overall assessment of the quality of Xanthii Herba and Xanthii Fructus.

LC-electrolyte switch in a contiguous time segments to analyze multi-components: Simultaneous determination of phenolic acids and iridoids in rat plasma after inhalation administration of Reduning aerosol

The optimization of electrolytes, kinds and concentrations, in mobile phase for multiple constituents analyzing using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS) was usually compromised to ensure good LC separation of partial components. However, the compromised electrolytes could lead to ionization suppression of some of the analytes. To solve the compromise of electrolytes within various components, taking phenolic acids and iridoids as a case, we used electrolyte switch in contiguous running time segments of UPLC-ESI-MS/MS to ensure chromatographic separation of chlorogenic acid, neochlorogenic acid and cryptochlorogenic acid and improve the response of geniposide. Then the method was applied for pharmacokinetic study of the four components in rat after inhaling Reduning aerosol for the first time. The complete separation of the three chlorogenic acid isomers was achieved and the LLOQs of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, and geniposide were 1, 1, 3, and 0.2 ng/mL, respectively. In conclusion, we developed a sensitive and time-saving LC-MS/MS method for the quantitative analysis of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, and geniposide in rat plasma, and this method appears to be useful for pharmacokinetic studies of Reduning aerosol. The method provided a sight to alleviate compromise of electrolytes in mobile phase for HPLC-ESI-MS in analyzing multi-components.

High-Performance Thin Layer Chromatography Method Development and Validation for Simultaneous Determination of Phenolic Acids in Selected Indian Bamboo Species

High-Performance Thin Layer Chromatography Method Development and Validation for Simultaneous Determination of Phenolic Acids in Selected Indian Bamboo Species

A New Method for Simultaneous Determination of Phenolic Acids, Alkaloids and Limonoids in Phellodendri Amurensis Cortex.

Phellodendri Amurensis Cortex (PAC) is a well-known herbal medicine in China with complex components, but the previous research has mostly focused on its alkaloids analysis. For the first time, a simpler and more efficient method was proposed in this paper to simultaneously determine the content of three different kinds of compounds—phenolic acids, alkaloids and limonoids—in PAC. The phenolic acids included 3-O-feruloylquinic acid, 4-O-feruloylquinic acid and syringin. The alkaloids include magnoflorine, phellodendrine, jatrorrhizine, palmatine and berberine, while the limonoids include obaculactone and obacunone. An approach combining multi-wavelength and HPLC-DAD was used in this study due to the great difference in maximum absorption wavelength of the various components. Four wavelengths at 215, 275, 280 and 310 nm, respectively, were chosen for monitoring. It has been indicated through appropriate tests that this approach is of high accuracy, good repeatability and stability and provides a scientific basis for the quality assessment of PAC and associated derivatives. In addition, the chromatographic fingerprints method combined with multivariate statistical analysis chosen in this study was proved to be effective and reasonable for an accurate classification of 33 batches of samples collected from different locations.

HPLC-ED Analysis of Phenolic Compounds in Three Bosnian Crataegus Species.

The aim of this work was the qualitative and quantitative determination of selected phenolic compounds in three Crataegus species grown in Bosnia. Crataegus plants are consumed for medicinal purposes and as foodstuff in the form of canned fruit, jam, jelly, tea, and wine. Two samples of plant material, dry leaves with flowers, and berries of three Crataegus species—Crataegus rhipidophylla Gand., Crataegus x subsphaericea Gand., and Crataegus x macrocarpa Hegetschw.—were analyzed. Twelve ethanolic extracts were isolated from the selected plant material using Soxhlet and ultrasound extraction, respectively. Soxhlet extraction proved to be more effective than ultrasound extraction. A simple and sensitive method, high-performance liquid chromatography with electrochemical detection, HPLC-ED, was used for the simultaneous determination of phenolic acids and flavonoids in Crataegus species. The content of gallic acid in the extracts ranged from 0.001 to 0.082 mg/g dry weight (DW), chlorogenic acid from 0.19 to 8.70 mg/g DW, and rutin from 0.03 to 13.49 mg/g DW. Two flavonoids, vitexin and hyperoside, commonly found in chemotaxonomic investigations of Crataegus species, were not detected in the examined extracts. In general, leaves with flowers samples are richer in gallic acid and rutin, whereas the berries samples are richer in chlorogenic acid. Distinct similarities were found in the relative distribution of gallic acid among the three species. Extracts of C. x macrocarpa had the highest content of all detected compounds, while significant differences were found in rutin content, depending on the plant organ. To the best of our knowledge, this is the first study reporting content of phenolic compounds in Crataegus rhipidophylla Gand., Crataegus x subsphaericea, and Crataegus x macrocarpa from Bosnia.

Simultaneous determination and qualitative analysis of six types of components in Naoxintong capsule by miniaturized matrix solid-phase dispersion extraction coupled with ultra high-performance liquid chromatography with photodiode array detection and quadrupole time-of-flight mass spectrometry.

A simple and effective sample preparation process based on miniaturized matrix solid-phase dispersion was developed for simultaneous determination of phenolic acids (gallic acid, chlorogenic acid, ferulic acid, 3,5-dicaffeoylqunic acid, 1,5-dicaffeoylqunic acid, rosmarinic acid, lithospermic acid, and salvianolic acid B), flavonoids (kaempferol-3-O-rutinoside, calycosin, and formononetin), lactones (ligustilide and butyllidephthalide), monoterpenoids (paeoniflorin), phenanthraquinones (cryptotanshinone), and furans (5-hydroxymethylfurfural) in Naoxintong capsule by ultra high-performance liquid chromatography. The optimized condition was that 25mg Naoxintong powder was blended homogeneously with 100mg Florisil PR for 4min. One milliliter of methanol/water (75:25, v/v) acidified by 0.05% formic acid was selected to elute all components. It was found that the recoveries of the six types of components ranged from 61.36 to 96.94%. The proposed miniaturized matrix solid-phase dispersion coupled with ultra high-performance liquid chromatography was successfully applied to simultaneous determination of the six types of components in Naoxintong capsules. The results demonstrated that the proposed miniaturized matrix solid-phase dispersion coupled with ultra high-performance liquid chromatography could be used as an environmentally friendly tool for the extraction and determination of multiple bioactive components in natural products.

Integrated approach for bioactive quality evaluation of medicinal plant extracts using HPLC-DAD, spectrophotometric, near infrared spectroscopy and chemometric techniques

ABSTRACTTo establish a universal analytical tool that could be used as a bioactive quality determination procedure on medicinal plant extracts, a range of spectrophotometric assays, HPLC, near infrared reflectance spectroscopy and chemometric analysis were employed for determination of the bioactive quality of 16 widely spread medicinal plants. Macro-constituents (total carbohydrates, soluble polysaccharides, proteins, amino acids) and secondary plant metabolites (total phenols, flavonoids, hydroxycinnamic acids, flavons, and flavonols) were determined, and HPLC method for the simultaneous determination of phenolic acids and flavonoids was developed, and its linearity, limits of detection and quantification, precision, and accuracy were validated. The evaluated medicinal plant extracts were characterised by a marked protein (marigold – 4.22 g/kg dw), amino acid (marigold – 61.14 g/kg dw), and carbohydrate content (dandelion – 113.5 g/kg dw), while Lamiaceae plants were distinguished as the predominant sources of polyphenolic bioactives (<2.26 g GAE/L). The developed HPLC method enabled separation of 24 polyphenolic compounds within a short analysis time (30 min) and revealed rosmarinic and chicoric acids as the prevalent polyphenolic constituents. NIR spectroscopy coupled with chemometric analysis of all determined analytical parameters indicated the suitability of NIR analysis for amino acids, carbohydrates, and polyphenols determination in medicinal plant extracts.

Simultaneous determination of phenolic acids and diterpenoids and their comparative pharmacokinetic study in normal and acute blood stasis rats by UFLC–MS/MS after oral administration of Guan-Xin-Shu-Tong capsules

Guan-Xin-Shu-Tong capsules are one of the well-known and first-line Chinese traditional herbal formula for treating coronary heart disease. A validated and sensitive method via ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) was established to simultaneously determinate five phenolic acids and four diterpenoids in rats in order to investigate their pharmacokinetic profiles firstly. Analytes were extracted by ethyl acetate and determined via multiple reaction monitoring mode in both positive and negative ion modes. The values for limit of quantification were in range of 0.025-1.250ng/ml. Inter- and intra-day precisions were no more than 10.9% with accuracy of -11.0%-10.6%, meanwhile the stable and suitable extraction recoveries were also obtained. And finally such excellent method was used to compare the pharmacokinetics of nine compounds in normal and acute blood stasis rats after oral administration of Guan-Xin-Shu-Tong capsules.

Simultaneous determination of phenolic acids and flavonoids in Chenopodium formosanum Koidz. (djulis) by HPLC-DAD-ESI–MS/MS

A high performance liquid chromatography-diode array detection-tandem mass spectrometry method (HPLC-DAD-MS/MS) was developed for simultaneous determination of phenolic acids and flavonoids in djulis (Chenopodium formosanum Koidz.), a traditional Chinese herb reported to possess vital biological activities. A high yield of phenolic acids and flavonoids was attained by employing 50% ethanol in water as the extraction solvent and shaking in a 60°C water bath for 3h. A total of 8 phenolic acids and 14 flavonoids were separated and identified within 55min by using a Poroshell 120 EC-C18 column with detection at 280nm, flow rate at 0.8mL/min, column temperature at 35°C, and a gradient solvent system of 0.1% formic acid in water and acetonitrile. Two internal standards caffeic acid and kaempferol-3-O-rutinoside were used for quantitation of phenolic acids and flavonoids in djulis respectively. The amounts of phenolic acids ranged from 11.5±0.8μg/g (caffeoyl-putrescine-derivative (2)) to 1855.3±16.9μg/g (hydroxylphenylacetic acid pentoside), while the flavonoids ranged from 19.93±2.29μg/g (quercetin-3-O-(coumaryl)-rutinoside-pentoside (1)) to 257.3±2.05μg/g (rutin-O-pentoside (2)). A high recovery (89.68–97.20%) and high reproducibility was obtained for both phenolic acids and flavonoids with the relative standard deviation (RSD) for the latter ranging from 0.09–8.22% (intra-day variability) and 0.80–8.48% (inter-day variability). This method may be applied to determination of both phenolic acids and flavonoids in food products and Chinese herbs.

Simultaneous determination of phenolic acids in Guanxinning injection by HPLC

This paper was aim to determine five phenolic acids, sodium danshensu (SD), protocatechuic aldehyde (PA), rosmarinic acid (RA), lithospermic acid (LA) and salvianolic acid B (SAB), in Guanxinning injection. In the test, Kromasil C18 column (4.6 mm x 250 mm, 5 µm) was adopted, with acetonitrile-3% formic acid solution as the mobile phase for gradient elution. The flow rate was 1 mL · min, the column temperature was 30 °C and the detection wavelength was 280 nm. According to the results of the test, SD, PA, RA, LA and SAB showed good linear relations between peak areas and sample sizes in 0.006 06-4.04 (r = 0.999 3), 0.006 15-4.10 (r = 0.999 4), 0.005 94-3.96 (r = 0.999 3), 0.006 06-4.04(r = 0.999 1) and 0.006 09-4.06 (r = 0.999 2) µg, respectively. The average recoveries (n = 6) were 98.9% (RSD 0.75%), 98.1% (RSD 1.2%), 100% (RSD 0.77%), 98.7% (RSD 1.7%), 102% (RSD 0.68%), respectively. The above 5 components were determined in 13 batches of samples by using the established method. The method was simple, accurate and highly reproducible that it could be used for quality control of the components in Guanxinning injection.

Chemometrics-based Approach in Analysis of Arnicae flos.

Introduction:Arnica montana flowers have a long history as herbal medicines for external use on injuries and rheumatic complaints.Objective:To investigate Arnicae flos of cultivated accessions from Bulgaria, Poland, Germany, Finland, and Pharmacy store for phenolic derivatives and sesquiterpene lactones (STLs).Materials and Methods:Samples of Arnica from nine origins were prepared by ultrasound-assisted extraction with 80% methanol for phenolic compounds analysis. Subsequent reverse-phase high-performance liquid chromatography (HPLC) separation of the analytes was performed using gradient elution and ultraviolet detection at 280 and 310 nm (phenolic acids), and 360 nm (flavonoids). Total STLs were determined in chloroform extracts by solid-phase extraction-HPLC at 225 nm. The HPLC generated chromatographic data were analyzed using principal component analysis (PCA) and hierarchical clustering (HC).Results:The highest total amount of phenolic acids was found in the sample from Botanical Garden at Joensuu University, Finland (2.36 mg/g dw). Astragalin, isoquercitrin, and isorhamnetin 3-glucoside were the main flavonol glycosides being present up to 3.37 mg/g (astragalin). Three well-defined clusters were distinguished by PCA and HC. Cluster C1 comprised of the German and Finnish accessions characterized by the highest content of flavonols. Cluster C2 included the Bulgarian and Polish samples presenting a low content of flavonoids. Cluster C3 consisted only of one sample from a pharmacy store.Conclusion:A validated HPLC method for simultaneous determination of phenolic acids, flavonoid glycosides, and aglycones in A. montana flowers was developed. The PCA loading plot showed that quercetin, kaempferol, and isorhamnetin can be used to distinguish different Arnica accessions.SUMMARY A principal component analysis (PCA) on 13 phenolic compounds and total amount of sesquiterpene lactones in Arnicae flos collection tended to cluster the studied 9 accessions into three main groups. The profiles obtained demonstrated that the samples from Germany and Finland are characterized by greater amounts of phenolic derivatives than the Bulgarian and Polish ones. The PCA loading plot showed that quercetin, kaemferol and isorhamnetin can be used to distinguish different arnica accessions.

Phenolic compounds involved in grafting incompatibility of Vitis spp: development and validation of an analytical method for their quantification.

Graft incompatibility of Vitis spp is an unresolved worldwide problem with important economic consequences. Grafting comprises a complex set of morphological and physiological alterations, in which the phenolic compounds seem to be strongly involved. Therefore, a detailed analysis and recognition of structural phenolic compounds diversity in the two partners of a Vitis graft is of great importance to evaluate their role as markers of graft establishment. To optimise a sample extraction method, and to develop and validate a high-performance liquid chromatography (HPLC) method for the simultaneous determination of phenolic acids and flavonols in the graft union so as to understand their behaviour in the metabolism of the scion-rootstock system, using compatible and incompatible combinations of a Syrah cultivar and two rootstocks (R110 and SO4). Sixty extracts of Vitis grafting tissues were prepared and analysed by HPLC for the qualitative and quantitative determination of their phenolic profile. Among the phenolic compounds identified in the samples, one benzoic acid (gallic acid), three cinnamic acids (caffeic acid, ferulic acid and sinapic acid) and two flavonols (catechin and epicatechin) are potentially suitable as markers of graft incompatibility. The method developed presents good performance and lends itself readily for application in routine analysis of the phenolic composition of Vitis grafting tissues to distinguish compatible and incompatible combinations in the graft callusing stage.

SIMULTANEOUS DETERMINATION OF ROSMARINIC ACID, SALVIANOLIC ACID B AND CAFFEIC ACID IN SALVIA LERIIFOLIA BENTH. ROOT, LEAF AND CALLUS EXTRACTS USING A HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH DIODE-ARRAY DETECTION TECHNIQUE

Salvia leriifolia Benth. (Lamiaceae) is a medicinal plant with analgesic, hypoglycemic and anti-inflammatory properties. Many of the beneficial effects of Salvia spp. are attributed to the phenolic compounds. The present study aimed to determine the amounts of rosmarinic acid, salvianolic acid B and caffeic acid in the roots, leaves and calli of S. leriifolia using a simple, rapid and efficient HPLC method. In addition, the effect of different combinations of 2,4-dichlorophenoxyacetic acid and kinetin–as growth regulators–on phenolic composition was evaluated. The results indicated that the 3 phenolic acids were well separated using the applied gradient HPLC system [C18 (5mm) column; phosphoric acid 1% and methanol as mobile phase]. Limit of detection and quantification values for rosmarinic acid, caffeic acid and salvianolic acid B were 0.2 and 0.5, 1.0 and 2.0, and 0.5 and 1.0mg/mL, respectively. Inter- and intra-day variations for rosmarinic acid, caffeic acid and salvianolic acid B were 2.8% and 3.4%, 2.4% and 3.5%, and 4.8% and 4.7%, respectively. Recovery values for the 3 phenolic acids ranged from 96.3% to 98.9%. Rosmarinic acid was the most frequent phenolic acid in the ethanolic extract of roots, leaves and calli of S. leriifolia (0.33, 4.65 and 6.39 mg/g dry extract, respectively) followed by salvianolic acid B (0.11, 0.13 and 0.10 mg/g) and caffeic acid (0.15, 0.20 and 0.07 mg/g). The highest dry weight (0.226 g) of callus was achieved in medium containing 2 mg/L 2,4-D and 1 mg/L KIN. To conclude, the present study indicated that rosmarinic acid is the major phenolic acid in the roots, leaves and calli of S. leriifolia, using a simple, rapid, efficient and reproducible HPLC method for the separation and simultaneous determination of phenolic acids. Moreover, the content of evaluated phenolics was found to be higher in the leaf extract vs. root and callus extracts of the plant.

Simultaneous determination of phenolic acids by UPLC–MS/MS in rat plasma and its application in pharmacokinetic study after oral administration of Flos Lonicerae preparations

The current study aims to investigate the pharmacokinetic study of five phenolic acids (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 3,5-dicaffeoylquinic acid and 3,4-dicaffeoylquinic acid) following oral administration of Flos Lonicerae preparations in rats. A rapid and sensitive ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed to simultaneously determine the five phenolic acids in rat plasma. After mixing with the internal standard (IS) tinidazole, plasma samples were pretreated by liquid–liquid extraction with ethyl acetate/n-hexane (9:1, v/v). The separation was performed on an Acquity UPLC BEH C18 column (100mm×2.1mm, 1.7μm) at a flow rate of 0.4mlmin−1, and acetonitrile/methanol (4:1, v/v)-0.4% formic acid was used as mobile phase. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source with positive ionization mode. All calibration curves had good linearity (r>0.991) over the concentration ranges of 0.74–378ngml−1 for neochlorogenic acid, 0.50–1030ngml−1 for chlorogenic acid, 1.9–250ngml−1 for cryptochlorogenic acid, 0.74–380ngml−1 for 3,5-dicaffeoylquinic acid, and 5.1–328ngml−1 for 3,4-dicaffeoylquinic acid. The intra-and inter-day precision were within 15% and the accuracy ranged from 86.2% to 114.1%.

Simultaneous Determination of Phenolic Acids and Scopoletin in Brandies Using Synchronous Fluorescence Spectrometry Coupled with Partial Least Squares

Brandy, a spirit drink produced from wine (grape), is rich in phenolic acids due to its maturation in wooden barrels. Phenolic acids play a significant role in defining the sensorial characteristics of wines and brandies, and therefore, it is very useful to determine them. Synchronous fluorescence spectra of mixtures containing phenolic acids (gallic, vanillic, syringic and ferulic) and scopoletin have been used for the determination of these compounds by partial least squares (PLS)2. Synchronous fluorescence spectra were collected by simultaneously scanning the excitation and emission monochromator in the excitation wavelength range 200–500 nm, with constant wavelength difference 100 nm between them. The leave-one-out cross-validation method was used to select the optimum number of five PLS2 components (latent variables). The PLS2 model captured for 100 % of variance in the spectral block, and it accounted for 99.34 % of variance in the concentration block. The performance of the model was evaluated by means of root mean square error of cross-validation, root mean square error of prediction and coefficient of determination. The best model was used for the determination of the above-mentioned compounds in brandy samples at concentration levels 2–74 mg L−1 for phenolic acids and 0.06–0.43 mg L−1 for scopoletin. The PLS2 results were found to be in good agreement with those obtained by HPLC method.

Simultaneous Determination of Phenolic Acids in Water Caltrop by HPLC-DAD

Trapa natans L., or the water caltrop, a native plant of Romania, has not been researched in our country until recently. It has, however, been used for the treatment of the several diseases since ancient times. The objective of this work is to validate the HPLC method to detect and quantify the phenolic acids from Trapa natans L. specie. The HPLC-DAD method was validated for the analysis of 3-O-methylgallic, chlorogenic, caffeic, and ferulic acids in water caltrop roots, leaves, pulp, and hulls. The analysis was conducted on a Zorbax XDB-C18 column with gradient elution of acetonitrile-orthophosphoric acid. The method was validated in terms of linearity, accuracy, precision, limit of detection, quantification, and recovery, and found to be satisfactory. We verified the performance parameters after experimental studies, and we established that the HPLC method can be applied for the determination of phenolic acids in plant materials.