Determination Of Total Flavonoids Research Articles

Quantitative determination of total flavonoids in bilberry shoots

The main flavonoid glycoside from bilberry (Vaccinium myrtillus L.) shoots was isolated and identified preliminarily as quercetin-3-O-xyloside. Amethod for quantitative determination of flavonoids in bilberry shoots based on differential spectrophotometry at analytical wavelength 420 nm with complexation by aluminum chloride was developed.

The antioxidant activity, total phenolics and total flavonoids content of Bryonia dioica Jacq.

Bryonia dioica Jacq. is used as a medicinal plant in traditional medicine in the west of Iran. In this study, we investigated antioxidant activities, total phenols and flavonoid of several extracts (polar and nonpolar) obtained from B. dioica Jacq. flowers, leaves and stems for two stages of growth (flowering and pre-flowering). Several complementary assays, radical scavenging capacity, determination of total flavonoids and reducing power were used to screen the antioxidant properties of extracts. Large differences in the composition of phenolic compounds were detected among flower, leaf, and stem in different stages of growth. In the light of the data obtained, B. dioica Jacq. flowers represent an important source of phytochemicals with potent antioxidant capacity.

Rapid Determination of Total Flavonoids in a Medicinal Plant Epimedium by Near-infrared Reflectance Spectroscopy

Rapid Determination of Total Flavonoids in a Medicinal Plant Epimedium by Near-infrared Reflectance Spectroscopy

A critical evaluation on the reliability of two aluminum chloride chelation methods for quantification of flavonoids

Flavonols kaempferol, quercetin, myricetin and gossypetin, and flavones apigenin, acacetin, luteolin, orientin and tricin, are subjected to two AlCl3 spectrophotometric methods used for determination of total flavonoid content. The method developed by Woisky and Salatino involves addition of AlCl3 solution to the flavonoid solution and recording of optical density at 420nm. All flavonols except kaempferol have absorption maxima above 440nm and so readings at 420nm are erroneous. Among flavones, all except for luteolin and orientin, have absorption maxima below 400nm. Further, addition of CH3COOK and recording the absorbance at 415nm, as modified by Chang et al., works well for flavonols kaempferol, quercetin and myricetin, but not for gossypetin. The flavones luteolin and orientin absorbed above 400nm, whereas all others absorbed below 400nm. Examination of the results of both methods indicates they are inadequate, and should not to be considered as universal and standard methods for total flavonoid determination.

Assessment of flavonoids contents and in vitro antioxidant activity of Launaea procumbens.

BackgroundLaunaea procumbens (LP) has been used as a food supplement in Pakistan. In this study methanolic crude extract (LPME) of the whole plant and its different fractions; n-hexane (LPHE); ethyl acetate (LPEE) and chloroform (LPCE) were studied for the determination of total flavonoid and phenolics contents along with multifaceted in vitro scavenging assays.ResultsConsiderable amount of flavonoid and phenolics contents were found in all the fractions. Methanol and chloroform fraction exhibited efficient scavenging of DPPH·, ABTS·+, ·OH, superoxide, lipid peroxide and nitric oxide free radicals. Significant correlation was found between DPPH·, ABTS·+, superoxide radical, β-carotene bleaching restraint and phosphomolybdenum assay with total flavonoids and phenolics contents. High performance chromatography (HPLC) of LPME revealed the presence of vitexin, orientin, rutin, hyperoside, catechin and myricetin.ConclusionThese results reveal the presence of bioactive compounds in LPME, which might be contributed towards the various in vitro scavenging.

Quantitative determination of total and individual flavonoids in stems and leaves of Buddleja davidii and Buddleja albiflora

Background:Buddleja davidii and B. albiflora are two different original plants of the famous crude medicine “Diaoyangchen”.Materials and Methods:An ultraviolet–visible spectrophotometric method and a HPLC method were used for the determination of total and individual flavonoids (luteolin and apigenin) contents from their stems and leaves for the first time.Results:From the comparative evaluation, remarkable differences in flavonoids contents were observed between different origins and different parts of the samples. And content of specific flavonoid did not correspond to the total flavonoids contents in Buddleja davidii and Buddleja albiflora.Conclusion:With a better accuracy and precision, the methods had been proved simple, rapid, and reliable for quantitative determination of the total flavonoids and luteolin and apigenin in the two phytomedicines. Furthermore, our present study will pave the way of guidelines for the differentiation and standardization and exploitation of individual parts of this herb material.

Validação de método e determinação espectrométrica dos flavonoides das folhas e do vinhoto da cana-de-açúcar e comparação com método CLAE-UV

This paper reports on a modification of the procedures originally described in the French Pharmacopoeia for the UV-visible spectrometric analysis of flavonoids, and proposes a validation of the method and its application in the determination of total flavonoids from sugarcane (Saccharum officinarum) leaves and vinasse. An analysis of precision and accuracy revealed a low relative standard deviation (< 5.0%) and a good recovery percentages (99.79 and 98.34%). A comparison of the spectrometric results against those obtained by high performance liquid chromatography (HPLC-UV) demonstrated complete compatibility between the modified French Pharmacopoeia (spectrometric) and HPLC-UV methods

Antimutagenic Potential ofSolanum lycocarpumagainst Induction of Chromosomal Aberrations in V79 Cells and Micronuclei in Mice by Doxorubicin

Solanum lycocarpum A. St. Hil. (Solanaceae) is a hairy shrub or small much-branched tree of the Brazilian Cerrado. S. lycocarpum fruits are commonly used in traditional medicine in powder form or as folk preparations for the treatment of diabetes and obesity, as well as for controlling cholesterol levels. The aim of the present study was to chemically characterize the hydroalcoholic extract (SL) of S. lycocarpum by determination of total flavonoids and total poyphenols and quantification of steroidal alkaloids, as well as to evaluate its mutagenic and/or antimutagenic potential on V79 cells and Swiss mice using chromosomal aberrations and bone marrow micronucleus assays, respectively. Three concentrations of SL (16, 32, and 24 µg/mL) were used for the evaluation of its mutagenic potential in V79 cells and four doses (0.25, 0.50, 1.0, and 2.0 g/kg body weight) were used for Swiss mice. In the antimutagenicity assays, the different concentrations of SL were combined with the chemotherapeutic agent doxorubicin (DXR). HPLC analysis of SL gave contents of 6.57 % ± 0.41 of solasonine and 4.60 % ± 0.40 of solamargine. Total flavonoids and polyphenols contents in SL were 0.04 and 3.60 %, respectively. The results showed that not only SL exerted no mutagenic effect, but it also significantly reduced the frequency of chromosomal aberrations induced by DXR in both V79 cells and micronuclei in Swiss mice at the doses tested.

Study on the content determination of total flavonoids in Olea europaea L. leaves

A quantitative determination method of the total flavonoids in Olea europaea L. leaves using rutin as a standard substance was developed. Two coloration systems including Al(NO3)3-NaNO2-NaOH and AlCl3 were both used. The latter was selected as the optimal determination method by comparing with the results. The main interference of the matrix was discussed. The result showed that the AlCl3 coloration method is simple, accurate and reproducible with good linear relationship, and it can be looked as a reference method for the determination of total flavonoids in herbal materials or natural products.

Essential oil of Centaurea pannonica (Heufel) Simonkai and antioxidant activity of the methanol extract

During the last few decades, an intensive search of the safety of synthetic food additives has been carried out and many of them have been found to possess toxic activity [1]. Therefore the general trend is to replace them in food processing by natural antioxidants. Essential oils or plant extracts are proposed as alternative sources for the preservation of food [2]. Centaurea pannonica, is an erect perennial growing wild plant belonging to the Asteraceae family. Its essential oil was isolated from air-dried aerial parts by hydrodistillation and analyzed by GC and GC/MS [3]. Forty five compounds were identified, representing 82.2% of total oil, with 9-octadecanoic acid (34.0%), caryophyllene oxide (8.0%) and spathulenol (6.0%) being the main compounds. The methanol extract of C. pannonica was investigated for its antioxidant potential by using five different assays, including: total antioxidant capacity, free radical scavenging (DPPH), inhibitory activity toward lipid peroxidation, Fe3+-reducing power and Fe2+-chelating ability. The content of total phenolics in the methanol extract was determined according to the Folin-Ciocalteu procedure [4] and calculated as 105.96mg±9.22 GA/g dry extract. Moreover a spectrometric method with aluminum chloride [5] was used for the determination of total flavonoids, which was estimated to be 111.36±10.03mg rutin (RU)/g dry extract. The results show a significant antioxidant activity of the investigated extract compared to standard antioxidant compounds, such as butylated hydrohytoluene (BHT), gallic acid (GA), ascorbic acid (AA) and α-tocopherol.

Acid Hydrolytic Method for Determination of Ginkgo Biloba Total Flavonoids in Rat Plasma by HPLC for Pharmacokinetic Studies

A simple, reliable, economical method was developed using HPLC with a diode-array detector for determination of total flavonoids in plasma after introvenous administration of ginkgo biloba extract. The method simultaneously detects quercetin, kaempferol, and isorhamnetin after acid hydrolysis and recalculation. The hydrolysis and extraction conditions were optimized in an orthogonal test. The specificity was tested by comparing the retention time, UV spectra, and peak purity indices with standards. The detection limits were 20 ng/mL for quercetin, 20 ng/mL for kaempferol, and 50 ng/mL for isorhamnetin. The calibration curve ranges were 75–2400, 71–2280, and 70–2240 ng/mL. The pharmacokinetic characteristics of ginkgo biloba flavonoids after venous administration of 50 mg/kg ginkgo biloba extract to rats were analyzed using a two-compartment model. The initial plasma concentration was 171.22 μg/mL. The half-life of flavonoids in the first compartment (distribution) was 0.07 h and at the second compartment (elimination) was 4.51 h, while the AUC (0-∞) was 1711.06 μg·min/mL. The apparent volume of distribution was 0.11 L/kg. The total body clearance is 10.52 mL/(min·kg). The result shows the method is suitable for pharmacokinetic studies.

Determination of total flavonoids in three Sedum crude drugs by UV-Vis spectrophotometry

A simple, rapid UV–Vis spectrophotometry method for the determination of total flavonoids in Sedum sarmentosum Bunge., S. lineare Thunb., and S. erythrostictum Migo. was developed, with a good linearity, precision, and stability. The detection wavelength was set at 500 nm, and an extraction solvent was optimized. Through the comparative study of multiple samples of the three plant drugs, their collected seasons and the habitats can be preliminarily ascertained, which may help to control the quality of the medicines to some extent.

Determinação espectrométrica dos flavonóides das folhas de Maytenus (Celastraceae) e de Passiflora (Passifloraceae) e comparação com método CLAE-UV

Este trabalho apresenta uma modificação dos procedimentos descritos nas Farmacopéias Francesa e Européia para a análise de flavonoides de Passiflora incarnata L., Passifloraceae, por espectrometria UV-Visível e propõe a sua aplicação na determinação dos flavonoides totais das folhas da espinheira-santa (Maytenus aquifolium Mart. e Maytenus ilicifolia (Schrad.) Planch., Celastraceae) e do maracujá (Passiflora edulis Sims. e Passiflora alata Curtis, Passifloraceae). Os resultados obtidos por espectrometria no UV-Visível foram comparados aos obtidos por cromatografia líquida de alta eficiência (CLAE-UV), encontrando-se resultados estatisticamente similares entre os métodos espectrométrico modificado da Farmacopéia Francesa e CLAE-UV.

Quantitative determination of the total content of flavonoids in Flavozid and Immunoflazid syrups

New syrup compositions Flavozid and Immunoflazid that contain 2% Proteflazid (ethanol extract of wild Deschampsia caespitosa L. and Calamagrostis epigeios L.) have been developed. The total content of flavonoids in both syrups is of the level of 4 – 6 gmL, which complicates the development of an analytical method. A new method for quantitative determination of flavonoids in the aforementioned syrups has been developed on the basis of differential UV spectrophotometry of their complexes with aluminum chloride. The proposed method is introduced into the Manufacturer’s Pharmacopoeic Article for the syrups. The existing formulation of Flavozid and Immunoflazid syrups is based on the active pharmaceutical ingredient Proteflazid, which was first developed and registered in Ukraine in 2001 and is the alcohol extract of wild Deschampsia caespitosa L. and Calamagrostis epigeios L. Proteflazid prepared using the original technology was recommended for use in medical practice as a composition with direct antiviral and immunomodulating action with properties of an interferon activator and DNA protease inhibitor in virus-infected cells [1 – 3]. The principal biologically active compounds of Proteflazid are flavonoids similar to quercetin (rutin), the molecules of which are based on a flavone oxygen-containing heterocycle. However, the mixture of Proteflazid flavonoids did not contain those that are frequently used as standards. Therefore, the development of a quantitative determination method for flavonoids in the syrup, where the content of Proteflazid and the corresponding flavonoids is 2% (or 4 – 6 gmL) of the starting extract, is of definite scientific interest. The goal of our work was to develop a method that would enable quantitative determination of total flavonoids, which absorb in the UV and visible spectral regions, using aluminum chloride as a complex-forming reagent. The glycoside species were converted to the aglycons by preliminary boiling of the syrup in the presence of HCl in order to increase the total amount of flavonoids in the analytes. Acid hydrolysis of the glucosides in the syrup was carried out in an excess of conc. HCl taken at 1% of the volume of the investigated syrup. The optimum conditions for acid hydrolysis with time were determined from the degree of conversion of the glycoside component into aglycons with TLC monitoring and UV spectrophotometric data. Aglycons from the syrup were extracted by ethylacetate. Solvent was vacuum distilled until production of a pasty residue that was dissolved in ethanol (96%). Studies of the optimum conditions for complexation using freshly prepared aluminum chloride (5%) in ethanol (96%) showed that increasing the volume of the solution to greater than 1 mL in the analyte did not improve the optical density parameters of the aluminum-chloride complex of the flavonoids. The maximum optical density was retained over 25 – 40 min at wavelength 422 nm. A solution with all components of the test solution with the exception of aluminum chloride was used as a reference to compensate for absorption at 420 – 424 nm of the starting extract before complex formation with aluminum chloride. Considering the complex flavonoid composition of the starting material, known standards from various flavonoid groups were used for the quantitative determination. However, the absorption maximum of the complex with aluminum chloride of none of them agreed with that for the com

Determination of total flavonoids in Scutellaria barbata D. Don by dynamic ultrasonic extraction coupled with on-line spectrophotometry

A dynamic ultrasonic extraction (DUE) coupled with on-line detection by spectrophotometry is proposed for the determination of total flavonoids (calculated against scutellarin) in Scutellaria barbata D. Don. The extraction was performed in a common self-made extraction vessel put in an ultrasonic bath and a peristaltic pump was used to deliver the extraction solvent. Several experimental parameters of DUE, including flow rate and concentration of extraction solvent, ultrasonic power and sample amount, were optimized. In this work an on-line method was provided with the advantages of minimum sample amount (5 mg), on-line monitoring the extraction process and time saving (12.5 min). Compared with off-line method, the proposed method would be more convenient for obtaining continuous measurements and rapid optimization of the extraction process.

Continuous determination of total flavonoids in Platycladus orientalis (L.) Franco by dynamic microwave-assisted extraction coupled with on-line derivatization and ultraviolet–visible detection

This paper describes a new method for the determination of total flavonoids in Platycladus orientalis (L.) Franco. The method was based on dynamic microwave-assisted extraction (DMAE) coupled with on-line derivatization and ultraviolet–visible (UV–vis) detection. The influence of the experimental conditions was tested. Maximum extraction yield was achieved using 80% aqueous methanol of extraction solvent; 80 W of microwave output power; 5 min of extraction time; 1.0 mL min −1 of extraction solvent flow rate. The derivatization reaction between aluminium chloride and flavonoid is one of the most sensitive and selective reactions for total flavonoids determination. The optimized derivatization conditions are as follows: derivatization reagent 1.5% aluminium chloride methanol solution; reaction coil length 100 cm; derivatization reagent flow rate 1.5 mL min −1. The detection and quantification limits obtained are 0.28 and 0.92 mg g −1, respectively. The intra-day and inter-day precisions (R.S.D.) obtained are 1.5% and 4.6%, respectively. Mean recovery is 98.5%. This method was successfully applied to the determination of total flavonoids in P. orientalis (L.) Franco and compared with heat reflux extraction. The results showed that the higher extraction yield of total flavonoids was obtained by DMAE with shorter extraction time (5 min) and small quantity of extraction solvent (5 mL).

Differential Pulse Polarographic Determination of Flavonoids, I: Total Flavonoids in Forsythia viridissima Lindl

AbstractA differential pulse polarographic method is described for the determination of total flavonoids expressed in terms of rutin which is obtained from the dried blossoms of Forsythia viridissima Lindl, cultivated in parks. Differential pulse polarograms of rutin show a distinct peak at ‐ 1.83 V (vs Ag/AgCl) at pH 6.0 Sörensen buffer. The detection limit found was 10 ppm and the response was linear up to 100 ppm with a relative standard deviation (at the 20 ppm level) of 0.75%.

Spectrophotometric method of determining total flavonoids and phenol carboxylic acids in the exploitable preparation tanacin and in the inflorescence of common tansy

The exploitable preparation tanacin represents a quantity of biologically active substances. In its composition are flavonoids [i, 2] and phenol carboxylic acids (caffeic, chlorogenic, isochlorogenic) while the main component is the flavonoid aglycone luteolin. To standardize the preparation it is necessary to develop methods for the quantitative determination of total flavonoids and phenol carboxylic acids in the substance and in the plant raw material.