Study on phenolic acids of Lonicerae japonicae Flos based on ultrahigh performance liquid chromatography-tandem mass spectrometry combined with multivariate statistical analysis

Lonicerae japonicae Flos, a traditional Chinese medicine, has the function of evacuating heat and detoxifying. To promote the optimization of Lonicerae japonicae Flos germplasms and improve the quality of medicinal materials, 55 batches of five Lonicerae japonicae Flos germplasms with the same origin were collected during different periods, a UHPLC-TOF-MS method was established, and 22 kinds of phenolic acids were found and qualitatively analysed. Seventeen phenolic acids were selected for quantitative analysis by UHPLC-QqQ-MS/MS, and the quantitative results were analysed by principal component analysis, orthogonal partial least squares-discriminant analysis, and partial least squares discriminant analysis. The contents of phenolic acids in periods S1–S6 were found to be significantly different. There were also significant differences in the accumulation of phenolic acids in Lonicerae japonicae Flos during different growth periods. Ferulic acid, 5-O-caffeoylquinic acid, and caffeic acid were determined to be important components to distinguish the different growth periods of Lonicerae japonicae Flos. There were significant differences in the phenolic acid content of different germplasms of Lonicerae japonicae Flos, and the total amount of 17 phenolic acids and total acids (chlorogenic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid) in “Hua Jin No. 6” was highest, so the quality of “Hua Jin No. 6” was better than that of the four other germplasms. In addition, chlorogenic acid methyl ester and caffeic acid were the smallest markers in combination to distinguish the five germplasms of Lonicerae japonicae Flos.     

Phytochemical composition,in vitro antioxidant activity and antibacterial mechanisms of Neolamarckia cadamba fruits extracts

Aqueous extracts of Neolamarckia cadamba fruits prepared at different maturity stages were used for the analysis of various phytochemicals, and their antioxidant and antibacterial activities were determined. Ripe fruit extract had highest phenolics (3.14 mM GAE/ g fruit extract) with caffeic acid, tannic acid, syringic acid and quercetin as major phenolic compounds. The ripe fruit extract showed lowest IC₅₀ values in DPPH radical scavenging assay (231.33 μg fruit extract/ mL), and highest ABTS radical scavenging activity (111.18 μM TEAC/g). Immature fruit extract showed lowest minimum inhibitory concentration against tested bacteria, and the antibacterial activity was probably due to membrane permeation, as was evident by leakage of genetic material and reduction in propidium iodide uptake by bacterium; and by inhibition of sugar and amino acid uptake. The appreciable amount of phenolic compounds and biological activities in the aqueous extracts of N. cadamba fruits suggests it's potential application as natural preservative.     

HPLC–QTOF–MS/MS-based rapid screening of phenolics and triterpenic acids in leaf extracts of Ocimum species and their interspecies variation

Species of genus Ocimum are traditionally used for their medicinal and flavoring properties. These are rich sources of essential oils and found as an ingredient in many Ayurvedic preparations and food products. Phenolics and triterpenic acids are the medicinally active compounds mainly concentrated in the leaves of Ocimum species. This study aimed to develop an efficient and reliable analytical method for the rapid screening and characterization of phenolics and triterpenic acids in the leaf extracts of 6 Ocimum species using high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (HPLC–ESI–QTOF–MS/MS). A total of 50 compounds were identified and characterized on the basis of their accurate MS and MS/MS information, out of which 23 compounds were confirmed by authentic standards. Identified compounds include 28 flavonoids, 4 propenyl phenol derivatives, 2 triterpenic acids, 11 phenolic acids, and 5 phenolic acid esters. The developed method was applied to study the interspecies variation of identified compounds. Significant variation in the distribution of identified phenolics and triterpenic acids was observed among studied Ocimum species. Hence, the established method provides an effective and reliable tool for screening and characterization of phytoconstituents in Ocimum species.     

Phenolic constituents from Wissadula periplocifolia (L.) C. Presl. and anti-inflammatory activity of 7,4′-di-O-methylisoscutellarein

This study reports the first phenolics from Wissadula genus (Malvaceae) and the anti-inflammatory activity of 7,4′-di-O-methylisoscutellarein. Using chromatographic methods, five phenolic compounds were isolated from aerial parts of Wissadula periplocifolia (L.) C. Presl. The compounds were identified as 4-hydroxybenzoic acid, 3-hydroxybenzoic acid, trans-cinnamic acid, tamgermanetin and 7,4′-di-O-methylisoscutellarein using spectroscopic methods. The flavone 7,4′-di-O-methylisoscutellarein showed anti-inflammatory activity by inhibiting neutrophils recruitment in a mice model of pleurisy and by decreasing significantly the production of cytokines IL-1β and TNF-α.     

Purification of 4-hydroxybenzoic Acid and 4-hydroxybenzaldehyde from Laminaria japonica Aresch Using Commercial and Monolithic Sorbent in SPE Cartridge

Phenolic compounds are one of the major compounds in Laminaria japonica Aresch. Solid-phase extraction was used to separate the two phenolic compounds (4-hydroxybenzoic acid and 4-hydroxybenzaldehyde). A new monolithic sorbent was developed and used successfully as a special sorbent in a solid-phase extraction process. The effects on different SPE cartridges using silica, C₁₈, and monolithic were compared. Quantitative analysis was carried out using a C₁₈ analysis column (5 µm, 150 × 4.6 mm). Good linearity was obtained for the two targets, ranging from 0.5 to 100 µg mL^?1 (r ²  > 0.999), with relative standard deviations <4.7%.     

Identification and in vitro antioxidant activities of phenolic compounds isolated from Cynoglossum cheirifolium L.

In an extensive search for bioactive compounds from plant sources, the quantitative and qualitative characterisation of the compounds present in Cynoglossum cheirifolium extracts was studied. Total phenolic and flavonoid contents were determined by spectrophotometric techniques. In vitro antioxidant and radical scavenging profiling was determined through DPPH^? scavenging activity and Ferric reducing antioxidant power (FRAP). Our study showed that leaves produce more phenolic compounds, followed by flowering aerial part. The butanolic fraction obtained from leaves extract exhibited the highest total phenolics (78.65 ± 3.58 mg GAE/g DW) and flavonoids (22.15 ± 4.66 mg CE/g DW). In contrast, this fraction displayed the highest DPPH^? scavenging ability with IC₅₀ values of 0.06 ± 0.003 mg/mL. The RP-HPLC-PDA analysis of phenolic compounds from leaves of C. cheirifolium lets to identify: rosmarinic acid, ferulic acid, caffeic acid, p-coumaric acid, syringic acid, sinapic acid and rutin. The obtained results indicate that this plant represent a valuable source of natural antioxidants.     

Optimization and single-laboratory validation study of a high-performance liquid chromatography (HPLC) method for the determination of phenolic Echinacea constituents

Three species of Echinacea (Echinacea purpurea, Echinacea angustifolia, and Echinacea pallida) are commonly used for medicinal purposes. The phenolic compounds caftaric acid, cichoric acid, echinacoside, cynarin, and chlorogenic acid are among the phytochemical constituents that may be responsible for the purported beneficial effects of the herb. Although methods for the analysis for these compounds have been published, documentation of their validity was inadequate as the accuracy and precision for the detection and quantification of these phenolics was not systematically determined and/or reported. To address this issue, the high-performance liquid chromatography method, originally developed by the Institute for Nutraceutical Advancement (INA), was reviewed, optimized, and validated for the detection and quantification of these phenolic compounds in Echinacea roots and aerial parts.     

Activated Carbon Adsorption of Some Phenolic Compounds Present in Agroindustrial Wastewater

Single solute and simultaneous experimental adsorption isotherms of three phenolic compounds: gallic acid, p-hydroxybenzoic acid and syringic acid, have been investigated at 20, 30 and 40°C, using a bituminous coal based activated carbon. Regardless of temperature, the capacity of the activated carbon used to adsorb these compounds presented the following order: syringic acid > p-hydroxybenzoic acid > gallic acid. The increase of temperature slightly favored the adsorption capacity of the phenolic compounds. In binary and ternary component adsorption, experimental data suggest that interactions between adsorbates improve the adsorption capacity of some of the phenolic acid compounds. On the contrary, at high organic concentrations, adsorbed gallic acid was partially removed from the activated carbon surface because of the presence of the other components.     

Two new phenolic compounds from the leaves of Alnus sibirica Fisch. ex Turcz.

Two new phenolic compounds, 4-O-glucopyranosyl-5-O-caffeoylshikimic acid (1) and 2,3-digalloyl oregonin (2), were isolated along with eight known phenolic compounds (3–10) from an 80% acetone extract of Alnus sibirica leaves. The chemical structures of these compounds were elucidated using 1D/2D nuclear magnetic resonance and high resolution-MS. The anti-oxidative activities of these compounds were determined by assaying their 1,1-diphenyl-2-picrylhydrazyl radical and nitroblue tetrazolium superoxide anion scavenging activity. All of the isolated phenolic compounds (1–10) exhibited potent anti-oxidative activities. In particular, 2 and 4, which are diarylheptanoids, and 10 which is ellagitannin exhibited excellent anti-oxidative activities with almost the same potency as that of the positive controls L-ascorbic acid and allopurinol.     

A new cinnamic acid glycoside from roots of Heracleum dissectum

From the roots of Heracleum dissectum Lebb., one new cinnamic acid glycoside derivative named dissectumoside (1), together with eight known compounds including three phenolics, three phenolic glycosides and two phenylpropanoic glycoside were isolated using various chromatographic methods. Among them compound 2–9 was isolated from the plant for the first time. Their structures were elucidated and identified on the basis of their physicochemical properties and by extensive analyses of NMR spectroscopy and high-resolution mass spectrometry. The results of triglyceride accumulation screening in 3T3-L1 cells showed that compounds 1, 5 and 9 exhibited significantly accelerating activities of adipogenesis in adipocytes.     

New Phenolic Compounds from the Roots of Lentil (Lens culinaris)

While lentil (Lens culinaris) seeds are phytochemically well characterized, very little is known about secondary metabolites from lentil roots. Our research on lentil roots led to isolation of five phenolic compounds and five group B soyasaponins. Their structures were established using NMR spectroscopy and mass spectrometry. Four phenolics have not been hitherto described in the literature: 4‐O‐β‐d ‐glucopyranosyl‐2‐methoxybenzoic acid, (αS)‐4,4′‐di‐O‐β‐d ‐glucopyranosyl‐α,2′‐dihydroxydihydrochalcone, (αS)‐4′‐O‐β‐d ‐glucopyranosyl‐α,2′,4‐trihydroxydihydrochalcone, and keto‐2‐hydroxyglycitein. The DPPH^? radical‐scavenging activity of the purified phenolic compounds was additionally evaluated.     

LC-DAD-ESI-MS/MS-based phenolic profiling of St John’s Wort Teas and their antioxidant activity: Eliciting infusion induced changes

Infusion-induced changes in the phenolics, antioxidant and colour propeties of St John’s wort (genus, Hypericum; specie, Hypericum perforatum L.) teas were studied for the first time. SJW teas prepared as three different infusions and coded as three (3?min), six (6?min) and twelve minutes (12?min). Investigation of phenolic compounds were performed by LC-DAD-ESI-MS/MS. A total of 18 phenolics including six chlorogenic acids, three phenolic acids, seven flavonoids and two naphtodianthrones were detected. It is worth noting that the phenolic profiles of St John’s wort teas were similar. However, the quantities of phenolic compounds individually and totally changed significantly for different infusion times. The highest total concentration was detected in 12?min (60.03?mg/L), followed by 6?min (54.81?mg/L) and 3?min (33.07?mg/L). The main difference for different infusion times was the hyperoside found as the most abundant phenolic only in 3?min samples. However, for 6?min and 12?minin infusions, chlorogenic acid was the most dominant phenolic compound. Similar to phenolics, antioxidant capacity of tea infusions showed an increasing trend with the extension of infusion time.     

Release of Polyphenols Is the Major Factor Influencing the Bioconversion of Rice Straw to Lactic Acid

In this study, we found that p-coumaric acid (p-CA), ferulic acid (FA), and condensed tannins were released from rice straw during saccharification. The presence of polyphenols prolonged the lag phase and lowered the productivity of lactic acid. p-CA was identified as a key inhibitor. Tannins had a lower inhibitory effect than p-CA; FA had little inhibitory effect. Acid, alkaline, and ball milling pretreatments elicited different levels of polyphenol release from rice straw. Due to the different levels of polyphenol release in the pretreatment step, the enzymatic hydrolysates contained different concentrations of polyphenols. Compared with fermentation with a synthetic medium, fermentation with the hydrolysates of ball-milled rice straw provided much lower productivity and yield of lactic acid due to the presence of polyphenols. Removal of these compounds played an important role in lactic acid fermentation. When rice straw was alkaline pretreated, the hydrolysates contained few phenolic compounds, resulting in high productivity and yield of lactic acid (1.8 g/L/h and 26.7 g/100 g straw), which were comparable to those in a synthetic medium. This indicates that there is a correlation between removal of phenolic compounds and efficiency in lactic acid fermentation.     

HPLC characterization of phenolics in lignocellulosic materials

Summary The application of HPLC with an electrochemical detector for the determination of phenolics in lignocellulosic materials is reported. The separation of phenolic acids and aldehydes (gallic acid, p-hydroxybenzoic acid, vanillic acid, p-coumaric acid, syringic acid, ferulic acid, vanillin, syringaldehyde and p-hydroxybenzaldehyde) on two different columns (reversed phase C6 and styrene-divinylbenzene PLRP-S) is shown. Chromatograms of phenolic compounds in neutral, basic and oxidative extracts of wheat straw treated with NaOH and white rot fungusStropharia rugosoannulata are reported along with quantitative results.     

Chemical characterization of leaves and calli extracts of Rosmarinus officinalis by UHPLC-MS

Rosmarinus officinalis L. (Lamiaceae) is an aromatic plant widely popular mainly due to its uses in traditional medicine as an anti-inflammatory, diuretic and antimicrobial, as well as in the prevention and treatment of diseases. These biological activities are mainly related to the presence of phenolic and terpenic compounds. This work reports a chemical profile analysis of extracts from leaves and calli of rosemary obtained by both pressurized liquid extraction and maceration. Chemical profiles were determined on calli extracts of 3, 6, 9, and 15 days of culture; chemical characterization and quantification of compounds was carried out using ultrahigh performance liquid chromatography-mass spectrometry. A total of 53 metabolites were identified in callus and 47 compounds in leaf extracts, of which 25 correspond to phenolic compounds, mainly flavonoids and flavones, 13 terpenes that include phenolic terpenes and one diterpenolactone, two glycosides which correspond to 6-O-caffeoyl-β-D-fructofuranosil-(2→1)-α-D-glucopyranoside and primulaverin, an aromatic compound identified as fenantrenone and a growth regulator 12-hydroxy jasmonic acid. These results showed that undifferentiated rosemary cells accumulate the same compounds identified mainly in highly specialized tissues such as leaves. The plant cell culture supply the possibility of developing biotechnological processes to obtain compounds of commercial interest.     

Phytochemical screening,antioxidant activity,total phenolic and total flavonoid contents of seven local varieties of Rosa indica L.

Rosa indica symbol of godness and beauty known for various healing power, has astringent, sedative, anti-inflammatory and antidepressant qualities. Standard methods were used for qualitative detection of phyto-compounds, and quantitative detection of antioxidants was done using DPPH radical scavenging assay, total phenolics and total flavonoids content were expressed in mg GAE/g dry weight and mg QE/g dry weight. Results revealed phyto-compounds presence in all varieties under study however maximum % inhibition was observed by R. indica var pink perfume (94 ± 0.6) with IC50 value 0.3376 ± 0.01 mg/mL. Highest phenolic and flavonoid content was observed in the leaves extract of R. indica var cardinal red, i.e. 3.3553 ± 0.11 (ethanol) mg of Gallic acid equivalents (GAE)/g dry weight and 3.736 ± 0.001(ethanol) mg of quercetin equivalents (QE)/g dry weight, respectively, at conc. 0.125 mg/mL. Our finding provides evidence that all varieties of rose contain medicinally important bioactive compounds and justifies their use for treatment of different diseases.     

Effects of Extraction Methods on Phenolic Content in the Young Bamboo Culm Extracts of Bambusa beecheyana Munro

Nowadays, many studies focus on the potential of bamboo as a source of bioactive compounds and natural antioxidants for nutraceutical, pharmaceutical, and food sources. This study is a pioneering effort to determine the total phenolic content, total flavonoid content and free radical scavenging activity, as well as the phenolic identification and quantification of Bambusa beecheyana. The study was conducted by using ethanol, methanol, and water for solvent extraction by applying cold maceration, Soxhlet, and ultrasonic-assisted extraction techniques. The results showed that Soxhlet and ultrasonic-assisted Bambusa beecheyana culm extracts had an increase in the extract’s dry yield (1.13–8.81%) but a constant p-coumaric acid (4) content (0.00035 mg/g) as compared to the extracts from the cold maceration. The ultrasonic-assisted extraction method required only a small amount (250 mL) of solvent to extract the bamboo culms. A significant amount of total phenolics (107.65 ± 0.01 mg GAE/g) and flavonoids (43.89 ± 0.05 mg QE/g) were found in the Soxhlet methanol culm extract. The extract also possessed the most potent antioxidant activity with an IC₅₀ value of 40.43 µg/mL as compared to the positive control, ascorbic acid. The UHPLC–ESI–MS/MS analysis was carried out on the Soxhlet methanol extract, ultrasonic-assisted extract at 40 min, and cold methanol extract. The analysis resulted in the putative identification of a total of five phenolics containing cinnamic acid derivatives. The two cinnamic acid derivatives, p-coumaric acid (4) and 4-methoxycinnamic acid (5), were then used as markers to quantify the concentration of both compounds in all the extracts. Both compounds were not found in the water extracts. These results revealed that the extract from Soxhlet methanol of Bambusa beecheyana could be a potential botanical source of natural antioxidants. This study provides an important chemical composition database for further preclinical research on Bambusa beecheyana.     

UHPLC-MS/MS phenolic profiling and in vitro antioxidant activities of Inula graveolens (L.) Desf

Inula graveolens (L.) Desf. is an annual aromatic herb which has various uses on alternative medicine in many region of the world. In this study, antioxidant activities of ethanol and water extracts of the plant leaves were determined by in vitro DPPH method and phenolic composition of the plant sample was determined by LC-MS/MS analysis. The results showed that chlorogenic acid, quinic acid, hyperoside, protocatechuic acid and quercetin were the major phenolic compounds among the 27 standard compounds. The significant antioxidant capacity of the plant might be related with the high abundance of phenolic compounds.     

Rapid isocratic HPLC analysis of caffeic acid derivatives from <Emphasis Type="Italic">Echinacea purpurea</Emphasis> cultivated in Iran

Cichoric acid and caftaric acid are the main phenolic compounds in Echinacea purpurea tops. The level of these phenolic compounds in E. purpurea extracts is affected by different factors such as seasonal variations, drying methods, extraction methods, and growing location of the plant. HPLC analysis of caffeic acid derivatives in extracts of Echinacea purpurea (Cultivar) aerial parts, produced by boiling water extraction and ethanol-water extraction methods, showed various levels of the derivatives. Our findings revealed that the Iranian cultivated E. purpurea had a high level of cichoric acid (3.5–5.7 %). Caftaric acid was also the main phenolic compound in E. purpurea tops (3.1–4.5 %). After 2 h of boiling water extraction, the level of cichoric acid was 5.7 %, whereas the level of this acid in 60:40 ethanol-water extraction did not exceed 3.9 %. Published in Khimiya Prirodnykh Soedinenii, No. 2, pp. 150–152, March–April, 2008.