Flavonoid Assay Research Articles

Artemisia kruhsiana leaf extract induces autophagic cell death in human prostate cancer cells.

Some species of Artemisia have been reported to induce apoptosis and autophagy, but little is known of the apoptotic and autophagic effects of the stems and leaves of Artemisia kruhsiana Bess. (AkB). This study was conducted to investigate the antioxidant and anti-autophagic effects of the methanol extracts of the stems (EAkBs) and leaves (EAkBl) of AkB on human prostate cancer PC-3 cells. The antioxidant effects of EAkBs and EAkBl were measured using in vitro total flavonoid and total phenolic assays and a free radical scavenging assay. The effects of EAkBl on cell viability, apoptosis, autophagy, intracellular reactive oxygen species (ROS) generation and protein expression levels were also investigated. EAkBl was found to induce apoptosis, autophagy, and intracellular ROS generation in PC-3 cells. In terms of protein levels, EAkBl reduced phospho (p)-protein kinase B (AKT)/AKT, p-mammalian target of rapamycin (mTOR)/mTOR, B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein (Bax) ratios, and the activations of beclin 1/β-actin and microtubule-associated protein 1A/1B-light chain 3 (LC3) II/LC3 I ratios in PC-3 cells. The results of this study indicate EAkBl has antioxidant and anticancer effects on prostate cancer cells, and that these effects are associated with suppressions of p-AKT, p-mTOR, Bcl-2, and Bax, and the activations of beclin 1 and LC3. Our results indicate EAkBl has potential as a treatment for prostate cancer.

High-performance thin-layer chromatography fingerprinting and anti-inflammatory and antinociceptive activities of Pyracantha coccinea M.Roem.: A laboratory-based study.

This study aimed to investigate the high-performance thin-layer chromatography (HPTLC) fingerprinting and in-vivo anti-inflammatory and antinociceptive activities of different leaf extracts (ethanolic extract, n-hexane, chloroform, and ethyl acetate) of Pyracantha coccinea M.Roem. plant. A total of one hundred and twenty-four Wistar rats for anti-inflammatory and antinociceptive tests (carrageenan and formalin tests, respectively) were treated with two doses of the ethanolic extract (100 and 300 mg/kg), two doses of other plant fractions (30 and 100 mg/kg), Diclofenac (25 mg/kg) as the positive control, and normal saline as the negative control group, by oral gavage route. HPTLC fingerprinting is used for assay of terpenoids, flavonoids, alkaloids, and antioxidant activity. Treatment of the animal with the ethanolic extract at doses of 100 and 300 mg/kg, both ethyl acetate and chloroform fractions at the dose of 30 mg/kg and 100 mg/kg decreased the pain score in the formalin test and paw edema caused by carrageenan relative to control group significantly. Moreover, these extracts reported the highest amounts of flavonoid contents. In conclusion, phytochemicals present in Pyracantha coccinea M.Roem. leaves have anti-inflammatory and antinociceptive activities. Future studies are needed to identify the compounds with the anti-inflammatory and antinociceptive potential present in the plant.

A simple high-performance liquid chromatography method for the assay of flavonoids in Ginkgo biloba Leaves

Objective: Ginkgo biloba leaves, as an herbal medicine or dietary supplement, have been widely used worldwide. In this study, an integrated analytical method was established for the comprehensive analysis of flavonoids in G. biloba leaves. Materials and Methods: A practical chromatographic method combining high-performance liquid chromatography fingerprint analysis and quantitation was used to simultaneously determine 11 flavonoids (6 flavonol glycosides and 5 biflavones) in G. biloba leaves from different regions. Results: A total of 11 characteristic peaks were identified accurately, and the similarity of fingerprints ranged from 0.944 to 0.996. Methodology validation revealed appropriate linearity ( R2 ≥ 0.9997), precision, repeatability, stability, and recovery. The total contents of the six flavonol glycosides and five biflavones were within the range of 2.142-8.378 mg/g and 3.759-5.675 mg/g in 19 batches of samples, respectively. Among them, two coumaroyl flavonol glycosides were the predominant components. Conclusions: In this study, a convenient and reliable approach was successfully employed for the comprehensive evaluation of flavonoids in G. biloba leaves, which also provided a reference for its quality standard.

Phytochemical screening, antioxidant and antilithiasis activities of Juncus acutus

The objective of this work was to perform a phytochemical screening of Juncus acutus and to evaluate its antioxidant activity and antilithiasis activity. Phytochemical screening has demonstrated the presence of catechin, tannins and alkaloids in the various extracts studied. The antioxidant activity of the methanolic and ethyl acetate extracts by ultrasound and maceration was evaluated by the method of DPPH, FRAP and phosphomolybdate. The ethyl acetate extract by ultrasound, the methanolic extract by maceration and ultrasound respectively showed IC 50 with DPPH of 1,449mg / ml, 1,535mg / ml and 4,771mg / ml and a reducing power was observed at concentrations of 419,561mgBHA /100g, 207,143mgBHA /100g and 142,20mgBHA /100g by FRAP and at concentrations of 839,470mgAAE/100g, 283,260mgAAE/100g and 141,018mgAAE/100g by the phosphomolybdate test. The determination of the total polyphenols of the ethyl acetate extract by ultrasound, the methanolic extract by maceration and by ultrasound presented respectively contents of 2,428 mg GAE / g, 1,960mg GAE / g and 1,172mg GAE / g, likewise, the flavonoid assay showed respectively concentrations of 14,469mg EQ/g, 6,466 mg EQ/g and 3,143 mg EQ/g. Antilithiasis activity was evaluated on carbapatite, cystine and uric acid stones. The mass loss of uric acid, carbapatite and cystine stones was respectively 42%, 7% and 16%.

Study of the Biological Activities of the Seeds of the Plant Ceratonia Siliqua L. Recovered in the Bejaia Region

Background: Ceratonia siliqua is a plant that belongs to the Fabaceae family. It is frequently used in our culinary and medical traditions to fight cholesterol, acute diarrhea and digestive disorders. Methods: The hydrogen atom or electron donation abilities of the corresponding extracts and some pure compounds were measured from the bleaching of the purple-colored methanol solution of 2, 20-diphenylpicrylhydrazyl (DPPH), studying the anti-inflammatory effect by measuring the volume of the edema of the paw that has received carrageenan 1%. Antimicrobial activity in vitro was screened by using disc diffusion and micro-dilution methods. Results: The determination of the polyphenols of the methanolic extracts of the seeds reveals the richness of our extracts in polyphenols is 30 mg of gallic acid / g of extract. The aqueous and methanolic extract flavonoid assay shows that they contain high levels of flavonoids with contents of 07 and 10 mg equivalent of quercetin / g of extract respectively. The aqueous and methanolic extracts of Ceratonia siliqua seeds from the Bejaia region could bring the stable free radical 2.2 diphenyl-1-picrylhydrazyl (DPPH) to yellow-colored diphenylpicrylhydrazine with respective IC 50 values of 0.7 mg/ml and 0.2 mg/ml. They exhibit lower antioxidant activity than ascorbic acid (0.038 μg / ml). Determination of anti-inflammatory activity revealed that our aqueous and methanolic extracts of Ceratonia siliqua L seeds are able to reduce edema up to 81.89%. The methanoid extracts of Ceratonia siliqua L. seeds have a medium antibacterial action against bacteria: E. coli (13 mm), Staphylococcus aureus (12 mm) and Pseudomonas aeruginosa (12 mm). On the other hand, the aqueous extracts of the seeds of Ceratonia siliqua L have a weak antibacterial action against the same bacteria with a diameter of inhibition of 9 mm. However, the aqueous extracts of Ceratonia siliqua L seeds are endowed with a very important inhibitory action against candida albicans and it is similar to that of the antibiotic, antifungal Econazole (1%). Conclusion: According to the results observed, the two extracts of the seeds and pods have a potential anti-free radical and antioxidant measured compared to the standard antioxidant used. The results of the antimicrobial activity carried out in vitro on the 3 bacterial strains indicate that the two methanolic extracts of this plant have low antibacterial activity.

Multimodal α-Glucosidase and α-Amylase Inhibition and Antioxidant Effect of the Aqueous and Methanol Extracts from the Trunk Bark of Ceiba pentandra.

Postprandial hyperglycemia and oxidative stress are important factors that worsen the health condition of patients with type 2 diabetes. We recently showed that extracts from Ceiba pentandra mitigate hyperglycemia in dexamethasone- and high diet/streptozotocin-induced diabetes. Herein, we evaluated the postprandial regulatory properties and the antioxidant effects of the aqueous (AE) and methanol (ME) extracts from the stem bark of Ceiba pentandra. The phytochemical analysis of AE and ME was performed using the LC-MS technique and the total phenolic and flavonoid assays. Both extracts were tested for their ability to inhibit superoxide anion (O2•ـ), hydrogen peroxide (H2O2), protein oxidation, alpha-amylase, and alpha-glucosidase activities. The mode of enzyme inhibition was also determined in a kinetic study. AE and ME were both rich in phenolic and flavonoid compounds. ME was 2.13 and 1.91 times more concentrated than AE in phenolic and flavonoid compounds, respectively. LC-MS allowed the identification of 5 compounds in both extracts. ME and AE inhibited O2•ـ with IC50 of 51.81 and 34.26 μg/ml, respectively. On H2O2, they exhibited IC50 of 44.84 and 1.78 μg/ml, respectively. Finally, they exhibited IC50 of 120.60 and 140.40 μg/ml, respectively, in the inhibition of protein oxidation induced by H2O2, while showing IC50 of 39.26 and 97.95 μg/ml on the protein oxidation induced by AAPH. ME and AE inhibited alpha-amylase with IC50 of 6.15 and 54.52 μg/ml, respectively. These extracts also inhibited alpha-glucosidase, demonstrating IC50 of 76.61 and 86.49 μg/ml. AE exhibited a mixed noncompetitive inhibition on both enzymes, whereas ME exhibited a competitive inhibition on α-amylase and a pure noncompetitive inhibition on α-glucosidase. These results demonstrate that ME and AE scavenge reactive oxygen species and prevent their effects on biomolecules. Besides, ME and AE inhibit carbohydrate digestive enzymes. These properties may contribute to reduce postprandial hyperglycemia and regulate glycemia in diabetic patients.

Arginase Inhibitory, Antioxidant Activity, Total Phenolic Content and Total Flavonoid Content of Ethyl Acetate Extract of Caesalpiniaturtuosa Roxb Stem Bark

Objective: The purpose of this study is to investigate arginase inhibition, antioxidant activity, total phenolic content and total flavonoid content of ethyl acetate extract of Caesalpiniaturtuosa Roxb. Material and method: stem bark of Caesalpiniaturtuosa Roxb was extracted using hexane, ethyl acetate and methanol subsequently. The ethyl acetate extract was fractioned. Then, the fractions were subjected to arginase inhibition, antioxidant activity, total phenolic content and total flavonoid assay. Correlation was considered by statistical analysis. Result: Out of eight fractions, two fractions have no activity. Two fractions (3 and 6) have strong activity in arginase with inhibition 90.72 % and 91.41% respectively. Fraction 3 and 6 have strong antioxidant activity with IC50 25.98 μg/mL and 48.01 μg/mL respectively. Statistical analysis shows arginase inhibitor activity was not related with antioxidant activity, total phenolic content and total flavonoid content in this plant. Conclusion: Activity in arginase inhibition of fraction from ethyl acetate extract of Caesalpiniaturtuosa Roxb are not related to antioxidant, total phenolic and flavonoid content.

Determination of Antioxidant Value and Chemical Groups in Selected Medicinal Plants Used for Conditions Associated with Herpes Simplex and Herpes Zoster Infections in Kakamega County, Kenya

Aims: To Determine antioxidant value and chemical groups in selected medicinal plants used for conditions associated with Herpes simplex and Herpes zoster infections in Mukhwa sub-location, Kakamega County, Kenya.
 Study Design: A qualitative ethnobotanical survey for plant identification and chemical analysis for antioxidant assay and chemical group detection.
 Place and Duration of Study: Plant samples were collected in Mukhwa sub-location in September 2014. Sample processing and chemical group detection was carried out at the Center of Traditional Medicine and Drug Research of Kenya Medical Research Institute. Antioxidant assay was carried out at the Department of Medical Laboratory Sciences of the Kenya Medical Training College.
 Methodology: All 12 Community Health Workers, comprising 7 females and 5 males, were interviewed for identification of plant species. Antioxidant assay was carried out using the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) reduction assay and detection of flavonoids, terpenoids, alkanoids, saponins and phenols carried out using physicho-chemical methods.
 Results: Caesalpinia decapetala, Garcinia buchananii and Entada abyssinica, were the most potent sources of antioxidant with the concentration giving 50% DPPH reduction (RSa50) of 50, 20 and 10 µg/ml, respectively. The most abundant chemical groups were; alkaloids in Schkuhria pinnata, terpenoids in E. abyssinica, flavonoids in G. buchananii, the latter also contained the highest amount of phenols.
 Conclusion: The findings of antioxidant and chemical groups in selected medicinal plants support their use for HIV conditions.

Antioxidant potential ethanolic extract of Glycine max (l.) Merr. Var. Detam and daidzein

Antioxidants in chemical terms are electron donors and antioxidants are biologically compounds that can overcome the negative effects of oxidants in the body such as damage to vital cells of the body. The balance between oxidants and antioxidants is very important. Antioxidant from natural sources are safer than synthetic antioxidants. Soybeans are functional food source that contains essential amino acids, vitamin e, saponins and are rich in antioxidants such as flavonoids, isoflavones and anthocyanins. The aim of this research is to investigate antioxidant potential of ethanolic extract of black soybean (EEBS) and its compound daidzein. The phytochemical screening assay evaluated by modified Farnsworth methods and to find out phenolic and flavonoid content, total phenols and flavonoids were tested. DPPH (2,2-diphenyl 1-pichylhydazy), ABTS (2,2’-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) reducing activity, FRAP (ferric reducing antioxidant power) activity and H2O2 scavenging activity assay were used to measure antioxidant activity. The analysis of phytochemical of EEBS exhibit the presence of saponins, alkaloids, tannins, steroids and triterpenoids and terpenoids. Total phenol and flavonoid assay showed the presence of phenols and flavonoids. Four assay of antioxidant activity showed that EEBS at highest concentration exhibit higher activity (%), and from three assays EEBS has higher antioxidant activity (expressed as IC50) than daidzein. Overall, three of the four antioxidant tests performed can be concluded that EEBS has better antioxidants activity than daidzein. Further research is needed regarding black soybean as a promising antioxidant resource.

Evaluation of In Vitro Solar Protection Factor (SPF), Antioxidant Activity, and Cell Viability of Mixed Vegetable Extracts from Dirmophandra mollis Benth, Ginkgo biloba L., Ruta graveolens L., and Vitis vinífera L.

The aim of this study was to validate a HPLC method for the assay of flavonoids in extracts obtained from natural sources, i.e., from Dirmophandra mollis Benth, Ginkgo biloba L., Ruta graveolens L., and Vitis vinífera L. The potential sun protecting effect, antioxidant activity, and cell viability of the extracts were also determined. Individual extracts (obtained from each individual species) and a mixed extract (containing the four extracts) were analyzed by the validated HPLC method for the identification of flavonoids and quantification of rutin and quercetin. An in vitro cell viability study was carried out using the neutral red method. The in vitro sun protection factor was determined by spectral transmittance and in vitro antioxidant efficacy was evaluated against DPPH, ABTS, and AAPH radicals. The HPLC method used for the identification and quantification of flavonoids in extracts exhibited linearity, precision, accuracy, and robustness. Detection and quantification limits were, respectively, 2.881 ± 0.9 μg·mL−1 and 0.864 ± 0.9 μg·mL−1 for quercetin, and 30.09 ± 1 μg·mL−1 and 9.027 ± 1.1 μg·mL−1 for rutin. All extracts did not affect cell viability at the evaluated concentration range and exhibited a sun protection effect and antioxidant activity. Among the evaluated extracts, Ginkgo biloba L. and the mixed extract depicted the most expressive antioxidant activity. The mixed extract exhibited sunscreen protection against ultraviolet A (UVA) and ultraviolet B (UVB) and a critical wavelength of 372.7 ± 0.1. Our results translate the enhanced flavonoids’ composition of the mixed extract, which may be a potential alternative over sunscreens and antioxidants in pharmaceutic/cosmetic formulations.

Unveiling the Unexplored and Critically Endangered Ilex khasiana for its Antioxidant Properties

Ilex khasiana Purk. is an evergreen tree of Aquifoliaceae family reportedly endemic to Khasi Hills, Meghalaya, India; but it is also seen in a localised area of Aizawl, Mizoram, India. There is a vast history about the genus Ilex with reference to their medicinal uses in different traditional practices such as, in the treatment of coronary heart diseases, hypertension, hyperlipemia and hepatitis. Interestingly, I. khasiana is relatively unknown and yet it is known to possess multiple medicinal characteristics. The antioxidant potential studies were conducted on leaf extract specimens from Mizoram, using petroleum ether (IKP), chloroform (IKC) and methanol (IKM). IKM had the highest total antioxidant activity with 76.42 ascorbic acid equivalent (AE) mg/g, followed by IKP with 44.27 AE mg/g, and then by IKC with 17.08 AE mg/g respectively. The reducing power assay showed a concentration-dependent activity against potassium ferricyanide. The total phenolic content was found to be 3.46 gallic acid equivalent (GAE) mg/g for IKM and 1.450 GAE mg/g for IKC respectively. The total flavonoid content was 41.9 quercetin equivalent (QE) mg/g for IKC and 30.8 QE mg/g for IKM. IKP did not show any activity in phenolic and total flavonoid assays.

Characterization and antioxidant assay of yellow frangipani flower (Plumeria alba) extract

Yellow frangipani flowers (Plumeria alba) are plants that commonly found in the tropics, including Indonesia. Frangipani utilization has not optimum and research about it has not many conducted. This study aims to characterize and test the antioxidant activity of frangipani flower extract. The extraction conducted by maceration method using n-hexane, ethyl acetate, and methanol as solvents. Characterization includes phytochemicals assay (alkaloids, flavonoids, terpenoids, phenolic hydroquinone, tannin-polyphenols, and saponins), total phenolic assay, and total flavonoid assay. Antioxidant test using DPPH radical scavenging assay. The results of the characterization showed that frangipani flower extract positively contained alkaloid, flavonoids, tannins compounds in all solvent phases. Terpenoids are positive in n-hexane and ethyl acetate extract, while saponins are positive in methanol extract. The total phenolic content of n-hexane, ethyl acetate, and methanol extract were 3.6 mgGAE/g; 27.74 mgGAE/g; and 35.23 mgGAE/g. The total flavonoid content of n-hexane, ethyl acetate, and methanol extract were respectively 0.8 mgQE/g; 12.18 mgQE/g; 19.9 mgQE/g. The strongest antioxidant activity is possessed by ethyl extract, followed by methanol extract, and the last are n-hexane extract. Keywords: Antioxidant; frangipani flower; phytochemical; total phenolic; total flavonoid

The Optimization of Pudding Formulation Using Noni (Morinda citrifolia L.) Seen from Antioxidant Content and Sensory Characteristics

Noni (Morinda citrifolia L.) is a fruit that known to be effective against many diseases. Its phenol and flavonoid contentacted as antioxidant, anticancer, antiinflamation and to fluent the blood vessel. Because of its content that rich in antioxidant, this research aim to utilized it as a food product. Pudding is one of food product that categorized as a dessert, which in the production does not require heat treatment, so the antioxidant can be maintained. The purpose of this experiment is to find the optimum formulation of Noni pudding seen from antioxidant content and sensory characteristics. The method of this research is total phenolic assay, total flavonoid assay and sensory evaluation from 50 random panelis age between 18-25 years old. The result showed that Noni fruit has total phenolic 0,13423 mg GAE/ml and the total flavonoid 0,11819 mg GAE/mL. The best formulation of pudding resulted from sensory evaluation was found with the addition 10% concentration of Noni extract 10%.

Antidiabetic Activity of leaf extract from three types of Mangrove Originating from Sambera Coastal Region Indonesia

This study designed to determine the antidiabetic profile ethanol extracts of leaf extract from three types of mangroves; Avicennia sp., Rhizopora sp, and Sonneratia sp. Extraction process used maceration with ethanol solvent. The phytochemical assayis carried out qualitatively including assay of alkaloids, flavonoids, steroids, terpenoids, phenolic compounds, tannins, and saponins. The method used for the antidiabetic test is the method of blood glucose tolerance. The results showed that ethanol extract of three types of mangrove leaves Avicennia sp., Rhizopora sp, and Sonneratia sp. containing secondary metabolite group alkaloids, flavonoids, steroids, terpenoids, phenolic compounds, tannins, and saponins. Ethanol extract of leaves of the three types of mangroves plants has antidiabetic activity. The ethanol extract of leaves of mangrove Rhizophora sp. with a dose of 200 mg/kg BB + glucose 2 g / kg (group P-2) showed the most excellent antidiabetic activity and lowering blood glucose levels by 31.27%. This study confirmed that three types of Mangrove from Sambera Coastal Region Indonesia showed the antidiabetic profile.

Anti-inflammatory property and gas chromatography-mass spectrometry analysis of ethyl acetate fraction of Crateva adansonii DC leaves

Aims: This study was designed to determine the most active anti-inflammatory fraction of C. adansonii leaves and characterize the bioactive compounds. Methods: Dried and pulverized C. adansonii leaves (CAL) were extracted with 70% methanol, followed by successive solvent partitioning into hexane (CALH), ethyl acetate (CALE), butanol (CALB) and aqueous (CALA) leaf fractions. In vitro antioxidant assays carried out on the extract/fractions were DPPH, hydrogen peroxide, ferric reducing antioxidant power (FRAP), nitric oxide scavenging, total antioxidant capacity (TAC), total phenol (TP) and flavonoid (TF) assays. In vitro anti-inflammatory assays investigated were heat-induced bovine serum albumin (BSA) denaturation and human red blood cell (HRBC) membrane stabilization against hypotonicity- induced hemolysis. Formaldehyde-induced inflammation model in rats was carried out, followed by paw edema measurement as well as serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities assays. The ethyl acetate (CALE) fraction was subjected to GC-MS analytical method. Results: CAL extract and fractions substantially inhibited BSA denaturation and stabilized HRBC membrane against hypotonicity-induced hemolysis, however CALE exhibited the highest activity. CALE also suppressed formaldehyde-induced rat paw edema and significantly (p

Study the Antioxidant Activity of Sage (Salvia Officinalis) Leaves Extract

This study was carried out to evaluate two type of sage (salvia officinalis) leaves extracts (Ethanolic and Water extracts) by using two methods (reducing power and chelating ability). So this study shows the high content of phenolic compound of ethanolic and water extract of sage (15.8 and 14.6) % respectively. The leaves of sag were locally obtained, twenty gram of ground material was extracted by 250 ml ethanol 95% or distilled water at boiling point, under reflux for one hr. The extractive was filtered and evaporated at 50 C° to the compete dryness. Total phenol assay, total flavonoid assay, reducing power assay, and chelating ability assay were observed in this study. Results revealed that the ethanoic extract had highest percentage of reducing power than water extract of sage leaves as so as the chelating ability percentage.

Effect of Sulfites on Antioxidant Activity, Total Polyphenols, and Flavonoid Measurements in White Wine.

Polyphenols content and antioxidant activity are directly related to the quality of wine. Wine also contains sulfites, which are added during the winemaking process. The present study aimed to evaluate the effects of sulfites on the assays commonly used to measure the antioxidant activity and polyphenols and flavonoids content of white wines. The effects of sulfites were explored both in the standard assays and in white wine. The addition of sulfites (at 1–10 μg) in the standard assays resulted in a significant, positive interference in the Folin–Ciocalteu’s assay used for polyphenols measurements and in both the Ferric Reducing Antioxidant Power and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation decolorization assays, which were used for antioxidant activity evaluation. A negative interference of sulfites (at 1–20 μg) was observed for the colorimetric aluminium-chloride flavonoids assay. The addition of sulfites to organic white wines (at 25–200 mg/L wine) clearly resulted in a significant overestimation of antioxidant activity and polyphenols content, and in an underestimation of flavonoids concentration. To overcome sulfite interferences, white wines were treated with cross-linked polyvinylpyrrolidone. The total polyphenols content and antioxidant activity measurements obtained after polyvinylpyrrolidone treatment were significantly lower than those obtained in the untreated wines. Flavonoids were expected to be higher after polyvinylpyrrolidone treatment, but were instead found to be lower than for untreated wines, suggesting that in addition to sulfites, other non-phenolic reducing compounds were present in white wine and interfered with the flavonoid assay. In view of our results, we advise that a purification procedure should be applied in order to evaluate the quality of white wine.

In Vitro Antioxidant Potential and Effect of a Glutathione-Enhancer Dietary Supplement on Selected Rat Liver Cytochrome P450 Enzyme Activity.

Background There is considerable evidence that many people take dietary supplements including those of herbal origin as an alternative therapy to improve their health. One such supplement, with an amalgam of constituents, is CellGevity®. However, the effect of this dietary supplement on drug-metabolizing enzymes is poorly understood, as it has not been studied extensively. Therefore, we investigated the effect of CellGevity dietary supplement on selected rat liver microsomal cytochrome P450 (CYP) enzymes, the most common drug-metabolizing enzymes. We also determined the total antioxidant potential of this dietary supplement in vitro. Methods To determine the antioxidant potential of CellGevity dietary supplement, 2,2-diphenyl-2-picryl-hydrazyl (DPPH), total phenolic, and flavonoid assays were used after initial preparation of a solution form of the supplement (low dose, LD; 4 mg/kg and high dose, HD; 8 mg/kg). Rats received oral administration of these doses of the supplement for 7 days, after which the effect of the supplement on selected liver CYP enzymes was assessed using probe substrates and spectroscopic and high-performance liquid chromatographic methods. Rats which received daily administration of 80 mg/kg of phenobarbitone and distilled water served as positive and negative controls, respectively. Results The IC50 value of the supplement 0.34 ± 0.07 mg/ml compared to 0.076 ± 0.03 mg/ml of the BHT (positive control). The total phenolic content of the supplement at a concentration of 2.5 mg/ml was 34.97 g gallic acid equivalent (GAE)/100 g while its total flavonoid content at a concentration of 2.5 mg/ml was 6 g quercetin equivalent (QE)/100 g. The supplement significantly inhibited rat CYP2B1/2B2 (LDT 92.4%; HDT 100%), CYP3A4 (LDT 81.2%; HDT 71.7%), and CYP2C9 (LDT 21.7%; HDT 28.5%) while it had no significant inhibitory effect on CYPs 1A1/1A2, CYP1A2, and CYP2D6. Conclusion CellGevity dietary supplement possesses moderate antioxidant activity in vitro and has an inhibitory effect on selected rat liver CYP enzymes, suggesting its potential interaction with drugs metabolized by CYP enzymes.

Comparison of spectrophotometric methods of total flavonoid assay based on complex formation with aluminum chloride as applied to multicomponent herbal drug angionorm

Objective: The objective was to compare the complex formation of flavones (luteolin, apigenin), flavonols (morin, quercetin, and rutin), flavanones (naringenin), flavanonols (dihydroquercetin), caffeic, and ferulic acids with aluminum ion using two spectrophotometric methods and the application of these methods to estimate the flavonoid content in the angionorm herbal product. Materials and Methods: Method 1 included direct complex formation with aluminum chloride, and Method 2 involved preliminary nitrozation and subsequent complex formation of nitroso-derivatives with aluminum chloride. 2,4-dinitrophenylhydrazine method was also tested. Results: The conjugation system increase in the chromophore structure of nitroso derivatives is reflected by the hyperchromic effect of all substances (from 1.4 to 4-fold). The largest bathochromic shift of nitroso derivatives was seen for rutin, luteolin, and dihydroquercetin (to 510–530 nm). The total flavonoids estimation in the extract of the four herbs mixture, which constitute angionorm preparation active ingredient, depends on the chosen reference substance (3.49% calculated as luteolin when measured by Method 1 or 8.71% by Method 2; 7.97% calculated as dihydroquercetin when measured by Method 1 and 4.14% by Method 2. Conclusion: The complex formation according to Methods 1 and 2 does not allow the identification of the selective spectral regions typical for the certain flavonoids subgroups and hydroxycinnamic acids. Neither method of complex formation is suitable for the assay of total flavonoids in unknown samples or for the flavonoid content comparison in the different herbal material.

English

The purpose of this study was to understand and investigate the phytochemical and antimicrobial effects of different fractions of the methanolic extract of Corchorus capsularis leaf. For the extraction of lipophilic compounds with the help of methanol solvent system, cold extraction process was used, where the grinded coarse powder of the leaf was soaked in methanol and kept for four days for proper extraction. After that, the extract was concentrated by using rotatory evaporator and dried in room temperature. Compounds in the extract were separated according to their polarity by using vacuum liquid chromatography (VLC) and solvents of different polarity N-hexane, benzene, diethyl ether, chloroform, dichloromethane. Later, thin layer chromatography (TLC), flavonoid assay, disk diffusion, inhibitory action in nutrient broth was done for different fractions of the methanolic extract. Diethyl ether, chloroform, dichloromethane fractions showed good anti-oxidant property but poor antibacterial activity against different types of microorganism. Chloroform fraction showed good inhibitory activity against Aspergilus niger (fungus) in nutrient broth.   Key words: Corchorus capsularis, vacuum liquid chromatography (VLC), thin layer chromatography (TLC), disc diffusion method, nutrient broth, Bangladesh.