scholarly journals Identification and Determination of Seven Phenolic Acids in Brazilian Green Propolis by UPLC-ESI-QTOF-MS and HPLC

Molecules ◽  
2019 ◽  
Vol 24 (9) ◽  
pp. 1791 ◽  
Author(s):  
Shengwei Sun ◽  
Meijuan Liu ◽  
Jian He ◽  
Kunping Li ◽  
Xuguang Zhang ◽  
...  

Brazilian green propolis is a complex mixture of natural compounds that is difficult to analyze and standardize; as a result, controlling its quality is challenging. In this study, we used the positive and negative modes of ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time of flight mass spectrometry in conjunction with high-performance liquid chromatography for the identification and characterization of seven phenolic acid compounds in Brazilian green propolis. The optimal operating conditions for the electrospray ionization source were capillary voltage of 3500 V and drying and sheath gas temperatures of 320 °C and 350 °C, respectively. Drying and sheath gas flows were set to 8 L/min and 11 L/min, respectively. Brazilian green propolis was separated using the HPLC method, with chromatograms for samples and standards measured at 310 nm. UPLC-ESI-QTOF-MS was used to identify the following phenolic compounds: Chlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, caffeic acid phenethyl ester (CAPE), and artepillin C. Using a methodologically validated HPLC method, the seven identified phenolic acids were then quantified among different Brazilian green propolis. Results indicated that there were no significant differences in the content of a given phenolic acid across different Brazilian green propolis samples, owing to the same plant resin sources for each sample. Isochlorogenic acid B had the lowest content (0.08 ± 0.04) across all tested Brazilian green propolis samples, while the artepillin C levels were the highest (2.48 ± 0.94). The total phenolic acid content across Brazilian green propolis samples ranged from 2.14–9.32%. Notably, artepillin C quantification is an important factor in determining the quality index of Brazilian green propolis; importantly, it has potential as a chemical marker for the development of better quality control methods for Brazilian green propolis.

2019 ◽  
Vol 9 (2) ◽  
pp. 138-143
Author(s):  
Tianyun Li ◽  
Xiling Dai ◽  
Yichen Li ◽  
Guozheng Huang ◽  
Jianguo Cao

Background:Stenoloma chusanum (L.) Ching is a Chinese traditional medicinal fern with high total flavonoid and total phenolic content. Traditionally, phenolic compounds were separated by using column chromatography, which is relatively inefficient. </P><P> Objective: This study aims to use an efficient method to separate natural products from S. chusanum by Medium-Pressure Liquid Chromatography (MPLC) and High-Speed Counter-Current Chromatography (HSCCC).Methods:In the present research, firstly, a sample (2.5 g) from the dichloromethane extract of S. chusanum was separated by MPLC. Next, fraction P5 was purified by HSCCC with a two-phase solvent system composed of hexane-ethyl acetate-methanol-water (HEMWat) at a volume ratio of 2:4:1:4 (v/v/v/v). </P><P> Result: Four phenolic acids were obtained and their structures were identified by means of NMR and ESI-mass analysis. They were identified as: 1) protocatechuic acid (34 mg, purity 90.1%), 2) syringic acid (66 mg, purity 99.0%), 3) p-hydroxybenzoic acid (5 mg, purity 91.2%) and 4) vanillic acid (6 mg, purity 99.3%).Conclusion:The combination of MPLC and HSCCC is a high-efficiency separation method for natural products. This is the first report with regard to the separation of four phenolic acids in one step by MPLC and HSCCC from S. chusanum (L.) Ching.


2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Jinli Zhang ◽  
Yan Ding ◽  
Haizhou Dong ◽  
Hanxue Hou ◽  
Xiansheng Zhang

Phenolic acid profiles and antioxidant activities of outer bran, coarse bran, and shorts from blue, black, and purple wheat were analyzed. Phenolic acids were mainly in the bound form in pigmented wheat bran fractions. Phenolic acid content decreased in the order of outer bran, coarse bran, and shorts for the three pigmented wheat varieties. HPLC analysis of phenolic extracts demonstrated that the bound form of phenolic acids contained more ferulic, isoferulic, and p-coumaric acids compared to their free counterparts. Among the three pigmented wheat varieties, the bran fractions from blue wheat contained higher bound phenolic acids than the other two pigmented wheat bran fractions, except for purple coarse bran. The blue wheat outer bran had the highest total bound phenolic acid of 3458.71 μg/g while the purple wheat shorts had the lowest of 1730.71 μg/g. The contribution of bound phenolic acids to the total phenolic content and antioxidant activity was significantly higher than that of free phenolic acids. Blue wheat bran fractions had the highest radical scavenging activity against DPPH∙ while those of purple wheat gained the highest ABTS∙+ scavenging activity. High correlations were observed between TPC and radical scavenging capacities for DPPH and ABTS (R2>0.85, P<0.05).


2020 ◽  
Vol 142 (2) ◽  
pp. 379-399
Author(s):  
Agnieszka Szopa ◽  
Paweł Kubica ◽  
Łukasz Komsta ◽  
Aleksandra Walkowicz-Bożek ◽  
Halina Ekiert

Abstract Agitated shoot cultures of two aronias, Aronia melanocarpa (Michx.) Elliott and Aronia arbutifolia (L.) Pers., were maintained on Murashige & Skoog medium (1 mg/l BA and 1 mg/l NAA), both with and without the addition of various biosynthetic precursors of phenolic acids and depsides (phenylalanine, cinnamic acid, benzoic acid and caffeic acid). Each substance was added in 5 concentrations (0.1–10 mmol/l), each concentration at two time points (at the beginning and on the 10th day of cultures). Twenty-four phenolic acids were determined in methanolic extracts of the biomasses collected after 20 days of growth cycles by means of HPLC method with DAD detection. The presence of seven compounds was confirmed in all the extracts—five depsides (neochlorogenic, chlorogenic, cryptochlorogenic, isochlorogenic and rosmarinic acids), and syringic and caffeic acids. The main metabolites in A. melanocarpa shoot extracts were isochlorogenic, chlorogenic and neochlorogenic acids (max. 249.88, 450.35, 192.16 mg/100 g DW). The main metabolites in A. arbutifolia shoot extracts were: chlorogenic, isochlorogenic and cryptochlorogenic acids (max. 361.60, 224.5, 526.2 mg/100 g DW). The largest total amounts of the compounds were confirmed in the cultures of both aronias after the addition of cinnamic acid (989.79 and 661.77 mg/100 g DW, respectively) and caffeic acid (854.99 and 1098.46 mg/100 g DW, respectively) at concentrations of 5 mmol/l on 10th day of growth cycles. These maximum amounts were 3.41, 3.42, 2.95 and 5.67 times higher, respectively, than in the control cultures. This is the first report documenting the high production of depsides in shoot cultures of black and red aronias after feeding with their biosynthetic precursors.


Antioxidants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 488
Author(s):  
Agnieszka Szopa ◽  
Michał Dziurka ◽  
Sebastian Granica ◽  
Marta Klimek-Szczykutowicz ◽  
Paweł Kubica ◽  
...  

Schisandra rubriflora is a dioecious, underestimated medicinal plant species known from traditional Chinese medicine. The present study was aimed at characterising the polyphenolic profile composition and the related antioxidant capacity of S. rubriflora fruit, stem and leaf and in vitro microshoot culture extracts. Separate analyses of material from female and male specimens were carried out. This study was specifically aimed at detailed characterisation of the contribution of phenolic compounds to overall antioxidant activity using ultra-high-performance liquid chromatography with a photodiode array detector coupled to electrospray ionization ion trap mass spectrometry (UHPLC-DAD-ESI-MS3) and a high-performance liquid chromatography-diode array detector (HPLC-DAD). Using UHPLC-DAD-ESI-MS3, twenty-seven phenolic compounds from among phenolic acids and flavonoids were identified. Concentrations of three phenolic acids (neochlorogenic, chlorogenic and cryptochlorogenic acids) and eight flavonoids (hyperoside, rutoside, isoquercitrin, guaijaverin, trifolin, quercetin, kaempferol, and isorhamnetin) were determined using HPLC-DAD using reference standards. The highest total phenolic content was confirmed for the stem and leaf extracts collected in spring. The contents of phenolic compounds of in vitro biomasses were comparable to that in the fruit extracts. The methanolic extracts from the studied plant materials were evaluated for their antioxidant properties using various in vitro assays, namely free radicals scavenging estimation using 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), ferric-reducing antioxidant power (FRAP) and cupric-reducing antioxidant capacity (CUPRAC) as well as QUick, Easy, New, CHEap, and Reproducible CUPRAC (QUENCHER-CUPRAC) assays. A close relationship between the content of polyphenolic compounds in S. rubriflora and their antioxidant potential has been documented.


Antioxidants ◽  
2019 ◽  
Vol 8 (9) ◽  
pp. 405 ◽  
Author(s):  
Chunhe Gu ◽  
Kate Howell ◽  
Frank R. Dunshea ◽  
Hafiz A. R. Suleria

Polyphenols are naturally occurring compounds found largely in fruits and vegetables. The antioxidant properties of these polyphenols including total phenolic content (TPC), total flavonoid content (TFC), tannin content, 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH), 2,2′-azinobis-(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS) scavenging abilities and ferric ion reducing antioxidant power (FRAP) were measured among sixteen (16) plant foods (mango, blueberry, strawberry, black carrot, raspberry, dark grapes, garlic, ginger, onion, cherry, plum, apple, papaya, peach, pear and apricot) by modifying, standardising and translating existing antioxidant methods using a 96-well plate reader. Eighteen targeted phenolic acids and flavonoids were characterised and quantified using high-performance liquid chromatography-photometric diode array (HPLC-PDA) and verified by modifying an existing method of liquid chromatography coupled with electrospray-ionisation triple quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS). While most of these compounds were accurately detected by the HPLC-PDA at a low concentration, a few polyphenols in low concentrations could be only be characterised using the LC-ESI-QTOF/MS method. Our results showed that mango possessed the highest overall antioxidant activity, phenolic acid and flavonoid content among the selected fruits. Factor analysis (FA) and Pearson’s correlation tests showed high correlations among ABTS, DPPH, FRAP and phenolic acids, implying the comparable capabilities of scavenging the DPPH/ABTS free radicals and reducing ferric ions from the antioxidant compounds in the samples. Phenolic acids contributed significantly to the antioxidant activities, and flavonoids contributed more to tannin content based on the correlations. Overall, methods modified and standardized in this study can provide better understanding of high throughput technologies and increase the reliability of antioxidant data of different plant foods.


2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Mohammed Moniruzzaman ◽  
Chua Yung An ◽  
Pasupuleti Visweswara Rao ◽  
Mohammad Nurul Islam Hawlader ◽  
Siti Amirah Binti Mohd Azlan ◽  
...  

The aim of the present study was to characterize the phenolic acids, flavonoids, and antioxidant properties of monofloral honey collected from five different districts in Bangladesh. A new high performance liquid chromatography (HPLC) equipped with a UV detector method was developed for the identification of the phenolic acids and flavonoids. A total of five different phenolic acids were identified, with the most abundant being caffeic acid, benzoic acid, gallic acid, followed by chlorogenic acid and trans-cinnamic acid. The flavonoids, kaempferol, and catechin were most abundant, followed by myricetin and naringenin. The mean moisture content, total sugar content, and color characteristics of the honey samples were 18.36 ± 0.95%, 67.40 ± 5.63 g/100 g, and 129.27 ± 34.66 mm Pfund, respectively. The mean total phenolic acids, total flavonoid content, and proline content were 199.20 ± 135.23, 46.73 ± 34.16, and 556.40 ± 376.86 mg/kg, respectively, while the mean FRAP values and DPPH radical scavenging activity were 327.30 ± 231.87 μM Fe (II)/100 g and 36.95 ± 20.53%, respectively. Among the different types of honey, kalijira exhibited the highest phenolics and antioxidant properties. Overall, our study confirms that all the investigated honey samples are good sources of phenolic acids and flavonoids with good antioxidant properties.


2011 ◽  
Vol 78 (2) ◽  
pp. 97-103 ◽  
Author(s):  
Stanisław Weidner ◽  
Monika Karolak ◽  
Magdalena Karamać ◽  
Ryszard Amarowicz

Grapevine seedlings<em> Vitis vinifera</em> were grown in a greenhouse under optimum conditions (soil moisture ca 70%) and under drought stress (soil moisture ca 35%). In addition, some of the plants subjected to drought underwent subsequent regeneration under optimum conditions. Drought stress caused accumulation of total phenolic compounds in grapevine roots, which may indicate that these compounds play an important role in the adaptation of roots to growth under stress conditions. Phenolic acids found in the roots occurred in the ester-bound form only. p-coumaric acid was present in the highest concentrations (6.2 to 10.5 µg/g fresh matter). The content of ferulic acid was lower, ranging from 2.4 to 4.6 µg/g fresh matter. The lowest concentration in grapevine roots was achieved by caffeic acid (2.4 to 2.9 µg/g fresh matter). The levels of p-coumaric and ferulic acids in roots rose significantly under the drought stress, while the concentration of caffeic acid increased during the post-drought recovery period. This may suggest that some of the phenolic acids protect plants under stress conditions. All the extracts from grapevine roots had antioxidative properties, but the antiradical activity of the extracts obtained from roots subjected to drought stress was inferior to the control. The same extracts were also characterised by depressed reducing power. The results imply that tolerance of grapevine to soil drought may be associated with the value of antioxidative potential in root tissues of these plants.


2011 ◽  
Vol 75 (3) ◽  
pp. 215-218
Author(s):  
Tadeusz Krzaczek ◽  
Monika Gawrońska-Grzywacz

The analysis of the fractions of free and bound, liberated by hydrolysis, phenolic acids in the inflorescences and rhizomes with roots of <em>Hieracium pilosella</em> L. was conducted by 2D-TLC and RP-HPLC methods. In the inflorescences fourteen phenolic acids were found: chlorogenic, gallic, protocatechuic, homoprotocatechuic, caffeic, gentisic, p-hydroxyphenylacetic, p-hydroxybenzoic, m-hydroxybenzoic, p-coumaric, syringic, vanillic, ferulic and salicylic. In the rhizomes with roots nine phenolic acids were identified: chlorogenic, gallic, homoprotocatechuic, caffeic, gentisic, p- hydroxyphenylacetic, p-hydroxybenzoic, syringic and vanillic. By means of the RP-HPLC method the contents of major acids were estimated which ranged from 0.01 mg% up to 139.50 mg%. Caffeic acid is the remarkably dominating constituent of both plant materials. Phenolic compounds may contribute to invasiveness and medicinal properties of this species.


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