scholarly journals Determination of catechins in green tea leaves by HPLC compared to spectrophotometry

2021 ◽  
Vol 19 (3(75)) ◽  
pp. 28-33
Author(s):  
Oleksandr Yu. Maslov ◽  
Mykola A. Komisarenko ◽  
Yulia S. Kolisnyk ◽  
Tatyana A. Kostina

Aim. To study the qualitative composition, the quantitative content of catechins in green tea leaves and compare the data obtained with those evaluated by spectrophotometry.Materials and methods. Green tea leaves used for the analysis were collected in Anhui Province, China. The extract for the HPLC analysis was obtained by the maceration method with 60 % ethanol twice in the raw material/extractant ratio of 1 : 20. In the case of the spectrophotometric analysis, green tea leaves were extracted with 70 % ethanol twice by the maceration method in the raw material/extractant ratio of 1 : 20. The analysis of the extract from green tea leaves was performed by high performance liquid chromatography using a Prominence LC-20 Shimadzu chromatographic system (Japan) with a SPD-20AV spectrophotometric detector, an Agilent Technologies Microsorb-MV-150 column (reversed phase, C18 modified silica gel, length – 150 mm, diameter – 4.6 mm, particles size – 5 μm). Substances in the extract were identified by comparing the retention time and the spectral characteristics of the test substances with the same characteristics of the reference standards. Spectrophotometric measurements were carried out using a UV-1000 single beam spectrophotometer (China) with the pair of S90-309Q quartz square cells.Results and discussion. Using high performance liquid chromatography 5 catechins were identified. Among them epigallocatechin-3-O-gallate (10.85 %) predominated, while catechin (0.61 %) had the lowest concentration. The total amount of catechins in green tea leaves was 30.56 and 24.79 % by HPLC and spectrophotometry, respectively. The F- and t-tests showed that there was no significant difference between the results of HPLC and spectrophotometry.Conclusions. The qualitative composition and the quantitative content of catechins have been determined in the extract from green tea leaves by high performance liquid chromatography and spectrophotometry. Both HPLC and spectrophotometric methods can be used to determine the total catechin content in green tea leaves. The high content of catechins makes the extract promising for further study and creation of new herbal medicinal products and dietary supplements. The results obtained will be used for standardization of green tea leaves and for future pharmacological research of its extract.

Author(s):  
O. Yu. Maslov ◽  
S. V. Kolisnyk ◽  
M. A. Komisarenko ◽  
E. Yu. Akhmedov ◽  
S. M. Poluian ◽  
...  

The aim of work is study qualitative composition and quantitative content of flavonoids and phenolic acids in green tea leaves. Material and methods. The object of the study was green tea leaves, which were collected in Anhui Province, China. The analysis of 60  % ethanolic extract from green tea leaves was performed by high-performance liquid chromatography using a Prominence LC-20 Shimadzu chromatographic system (Japan) with an SPD-20AV spectrophotometric detector, an Agilent Technologies Microsorb-MV-150 column (reversed-phase, C18 modified silica gel, length – 250 mm, diameter – 4.6 mm, particles size – 5 μm). Identification of substances in the extract was carried out by comparing the retention time and the spectral characteristics of the test substances with the same characteristics of the reference standards. Results. 13 compounds were identified and determined by high-performance liquid chromatography. Among flavonoid aglycones quantitatively dominated by quercetin (0.35 %), in the case of flavonoid glycosides, it was luteolin-6-C-glycoside (1.30 %) and among phenolic acids, it was gallic acid (5.21 %). Conclusions. The qualitative composition, quantitative content of flavonoids and phenolic acids in the green tea leaves were determined by high-performance liquid chromatography. According to HPLC, the content of flavonoids in green tea leaves was higher than the content of phenolic acids.


2021 ◽  
Vol 19 (1(73)) ◽  
pp. 42-47
Author(s):  
U. Umarov ◽  
S. V. Kolisnyk ◽  
O. V. Kolisnyk ◽  
M. Fatkhullaeva ◽  
N. K. Chinibekova ◽  
...  

Aim. To study the qualitative composition and quantitative content of the polyphenolic compounds in Pimpinella anisum herb and determine their antioxidant activity.Results and discussion. Among the polyphenolic compounds contained in anise herb, chlorogenic acid(4.409 mg/g) predominates. Signifcant amounts of catechins (3.104 mg/g), apigenin derivatives (3.077 mg/g) and luteolin (1.864 mg/g) also accumulate in the herb. Minor amounts of myricetin (0.105 mg/g) and naringenin (0.019 mg/g) derivatives, rutin (0.189 mg/g), quercetin (0.028 mg/g), apigenin (0.009 mg/g) and hesperetin (0.002 mg/g) are present. According to the research results, the antioxidant activity of polyphenolic compounds of anise herb with reference to ascorbic acid was found to be 67.76 ± 0.05 mmol/g. Rutin exhibited the antioxidant activity at the level of 3979.59 ± 0.08 mmol/g.Experimental part. Pimpinella anisum herb collected during the flowering stage in the summer of 2019 inthe Kharkiv region (Ukraine) was used for analysis. The analysis of 70 % ethanolic extract from anise herb was performed by high performance liquid chromatography using a Prominence LC-20 Shimadzu chromatographic system (Japan) with a SPD-20AV spectrophotometric detector, an Agilent Technologies Microsorb-MV-150 column (reversedphase, C18 modifed silica gel, length – 150 mm, diameter – 4.6 mm, particles size – 5 μm). Identifcation of substances in the extract was carried out by comparing the retention time and the spectral characteristics of the test substances with the same characteristics of the reference standards. The antioxidant activity was determined by the potentiometric method (pH meter – Hanna 2550, with redox electrode EZDO PO50) with reference to ascorbic acid.Conclusions. The qualitative composition and quantitative content of polyphenolic compounds in the ethanolic extract of anise herb have been determined by high performance liquid chromatography. The total content of polyphenolic compounds is 17.576 mg/g. The antioxidant activity of polyphenolic compounds of anise herb with reference to ascorbic acid has been found to be 67.76 ± 0.05 mmol/g.Key words: polyphenolic compounds; anise; herb; high performance liquid chromatography; antioxidant activity


2021 ◽  
Vol 19 (1(73)) ◽  
pp. 16-24
Author(s):  
S. Yu. Sheiko ◽  
A. S. Shalamay

The development of medicines based on alder cone extracts led to the introduction of Altan and Altabormedicines into medical practice. The technology of extraction cake from cones has made it possible to obtain extracts with different therapeutic properties.Aim. To develop an effective method for studying the qualitative composition of the Altabor substance anddetermine the quantitative content of its components.Results and discussion. The Altabor substance is a complex mixture of ellagitannins containing more than 70 components. The main components of the extract (or their isomers) were determined by mass spectrometry and by comparing the retention times with the literature data. Gallic, ellagic, valoneic acids dilactone were conclusively determined by adding reference standards of these acids to the extract. The substance contains the following compounds: 2,3-hexahydroxydiphenoyl-(α/β)-glucose tr1 = 0.55 min, tr2 = 0.89 min (α and β isomers), 4,6-O-[(S)-valeonyl]-D-glucose (isomer) tr = 0.64 min, gallic acid tr = 1.198 min, pedunculagin tr1 = 3.63 min, tr2 = 4.62 min(α and β isomers), proecoxin A (isomer) tr = 4.78 min, valoneic acid dilactone tr = 6.19 min, ellagic acid pentoside (isomer) tr = 7.07 min, ellagic acid tr = 7.335 min.Experimental part. The composition analysis was performed using an Agilent 1200 chromatograph with a UV detector, a G6140 mass detector, an Alltech 3300 light scattering detector (ELSD), as well as the Agilent ChemStation Rev.B.04.03 software. The molecular weights of the compounds in the extract were determined using the method of mass spectrometry of ESI-electrospray ionization. The determination of the components was performed using an ultraviolet detector at a wavelength of 280 nm. The column was Rapid Resolution HT Cartige, 4.6 × 30 mm, 1.8 μm, Zorbax SB-C18.Conclusions. A new effective method of analysis of the Altabor substance has been developed; it allowsdetermining the qualitative and quantitative content of its structural components. The method gives the possibility to control the process of obtaining the Altabor substance, study the dependence of its composition on the conditions of its obtaining, batch number, place, time of the natural raw material collection, and study the composition of other pharmaceutical substances, the plant raw material containing tannins. The advantage of the method isthe short time (up to 10 min) of analysis using high-performance liquid chromatography at high resolution.Key words: gallotannins; ellagitannins; Altabor; chemical composition; structure; HPLC (high-performance liquid chromatography)


2015 ◽  
Vol 17 (4) ◽  
Author(s):  
L. H. Shostak ◽  
S. M. Marchyshyn ◽  
M. I. Lukanyuk ◽  
O. L. Demydyak

<p>There were determined the amino acids contents and complex of polysaccharides of the above and underground parts of Primula veris L. In the plant materials it was established the general content of amino acids and the content of free amino acids by high performance liquid chromatography. Ther were identified from 21 to 23 amino acids and 7 were essential. It was determined their quantity content. The fractions of soluble polysaccharides and pectins were isolated from the studied materials and determined their contents. In the leaves, flowers and rhizomes with<br />roots there were established the qualitative composition and quantitative content of the free saccharides by the gas chromatography.</p>


2019 ◽  
Vol 16 ◽  
Author(s):  
Joanna Wittckind Manoel ◽  
Camila Ferrazza Alves Giordani ◽  
Livia Maronesi Bueno ◽  
Sarah Chagas Campanharo ◽  
Elfrides Eva Sherman Schapoval ◽  
...  

Introduction: Impurity analysis is an important step in the quality control of pharmaceutical ingredients and final product. Impurities can arise from drug synthesis or excipients and even at small concentrations may affect product efficacy and safety. In this work two methods using high performance liquid chromatography (HPLC) were developed and validated for the evaluation of besifloxacin and its impurity synthesis, with isocratic elution and another with gradient elution. Method: The analysis by HPLC in isocratic elution mode was performed using a cyano column maintained at 25 °C. The mobile phase was composed by 0.5% triethylamine (pH 3.0): acetonitrile (88:12 v/v) eluted at a flow rate of 1.0 ml/min with detection at 330 nm. The gradient elution method was carried out with the same column and mobile phase components only modifying the rate between organic and aqueous phase during analysis. The procedures have been validated according to internationally accepted guidelines, observing results within acceptable limits. Results: The methods presented were found to be linear in the 140 to 260 µg/ml range for besifloxacin and 0.3 to 2.3 µg/ml for an impurity named A. The limits of detection and quantification were respectively 0.07 and 0.3 µg/ml for impurity A, with a 20 µL injection volume. The precision achieved for all analyses performed provided RSD inter-day equal to 6.47 and 6.36% for impurity A with isocratic elution and gradient, respectively. The accuracy was higher than 99% and robustness exhibited satisfactory results. In the isocratic method an analysis time of 25 min and 15 min was obtained for gradient. For impurity A, the number of theoretical plates in the isocratic mode was about 5000 while in the gradient mode it was about 45000, hence, it made the column more efficient by changing the mobile phase composition during elution. In besifloxacin raw material and in pharmaceutical product used in this study, other related impurities were present but but impurity A was searched for and not detected Conclusion: The proposed methods can be applied for quantitative determination of impurities in the analysis of the besifloxacin raw material, as well as in ophthalmic suspension of the drug, considering the quantitation limit.


Fitoterapia ◽  
2020 ◽  
Vol 3 (3) ◽  
pp. 44-50
Author(s):  
S.M. Marchyshyn ◽  
◽  
L.V. Slobodianiuk ◽  
R.Yu. Basaraba ◽  
N.А. Hudz ◽  
...  

Keywords: amino acids; common pussytoes; yacon; stevia; golden marigolds; leaves; herb; aster family; high performance liquid chromatography. For the first time the qualitative composition was studied and the quantitative content of amino acids in the herb of common pussytoes, yacon leaves, stevia leaves, herb of golden marigolds was determined by HPLC. 17 bound and 16 free amino acids were identified in the herb of common pussytoes, in the herb of golden marigolds, yacon leaves and stevia leaves – 12 and 5, 17 and 11 and 16 and 14 amino acids, respectively. The highest content of the sum of essential and substitute amino acids is contained in the leaves of stevia; the lowest content of the sum of essential amino – in the herb of golden marigolds, the sum of essential amino acids – in the herb of common pussytoes.


2014 ◽  
Vol 32 (1) ◽  
pp. 227-232 ◽  
Author(s):  
M.A.P. Oliveira ◽  
U.R. Antuniassi ◽  
E.D. Velini ◽  
R.B. Oliveira ◽  
J.F. Salvador ◽  
...  

The objective of the present study was to analyze the influence of spray mixture volume and flight height on herbicide deposition in aerial applications on pastures. The experimental plots were arranged in a pasture area in the district of Porto Esperidião (Mato Grosso, Brazil). In all of the treatments, the applications contained the herbicides aminopyralid and fluroxypyr (Dominum) at the dose of 2.5 L c.p. ha-1, including the adjuvant mineral oil (Joint Oil) at the dose of 1.0 L and a tracer to determine the deposition by high-performance liquid chromatography (HPLC) (rhodamine at a concentration of 0.6%). The experiment consisted of nine treatments that comprised the combinations of three spray volumes (20, 30 and 50 L ha-1) and three flight heights (10, 30 and 40 m). The results showed that, on average, there was a tendency for larger deposits for the smallest flight heights, with a significant difference between the heights of 10 and 40 m. There was no significant difference among the deposits obtained with the different spray mixture volumes.


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