Quantification Of Tannins Research Articles

Diversity Of Physical Grain Quality Traits In Tropical Sorghum Genotypes

The current study envisioned to assess the physical traits of sorghum grain for selected sorghum genotypes. Seed for the genotypes was sourced from the Lupane State University Gene Bank and grown during the 2022/23 agricultural season at Lupane State University Farm experimental plots. At maturity, laboratory tests on kernel/grain hardness, 100 kernel weight, bulk density, kernel diameter, colour and determination of presence of tannins through qualitative tests were done for all the 24 sorghum genotypes. Results from analysis of variance demonstrated highly significant differences (P<0.001) on kernel weight, kernel diameter, kernel hardness and grain hardness showing a great diversity of physical traits among all the 22 genotypes and 2 commercial varieties of sorghum. Mean 100 kernel weight was 2.59g, kernel diameter was 3.49mm, bulk density was 1.23g/cm3 and kernel hardness was 28.9%. Visual assessment was done on grain colour and seed was classified under red, cream, white and brown sorghums, and mixed colours. A chi-square test found a significant relationship between grain colour and presence of tannins. Genotypes NPGRC3124, IS9405 showed moderate levels of tannins while IS13996, IS29925, NPGRC1699, NPGRC1156 and NPGRC1478 had high levels. A highly significant strong positive correlation was shown for sorghum genotype between kernel diameter and kernel weight (r=0.81 at p≤0.05). Highly significant positive correlation was also observed between bulk density and kernel weight (r=0.4173 at p≤0.001). Kernel hardness has a strong positive correlation with bulk density (r=0.6242). Quantification of tannins is recommended to prevent negative effects on human and livestock health.

Process optimization of microwave-assisted aqueous extraction of tannins and saponins from Malabar Spinach (Basella alba) leaves using ANN-GA and RSM methodology

The effect of microwave-assisted aqueous extraction (MAE) conditions on the recovery and quantification of tannins and saponins from Basella alba leaves has been explored. The experiment was carried out according to the Box-Behnken experimental design of response surface methodology (RSM) with 17 runs. The independent variables of the study were microwave power (100 to 300 W), treatment time (5 to 15 min), and feed/solvent ratio (0.025 to 0.05). The effect of extraction conditions on the response variables, total condensed tannin, total hydrolyzed tannin, and total saponin content, were predicted using RSM and artificial neural network-genetic algorithm (ANN-GA) methodology. The extracted values of total condensed tannin, total hydrolyzed tannin, and total saponin content, using RSM and ANN-GA were compared and evaluated for goodness of fit. The predicted values using RSM and ANN-GA models for total condensed tannin were found to be 0.18 and 0.26 mg TAE g−1, total hydrolyzed tannin were 0.14 and 0.15 mg TAE g−1, and total saponin content were 1.66 and 1.49 mg DE g−1, respectively. When compared to RSM, ANN-GA demonstrated higher value of R2 and lower values of other statistical parameters. Additionally, the predicted value of ANN-GA was more closely aligned with the experimental value.

Evaluation of free radical quenching, anti-inflammatory activity together with anticancer potential of Lychnis coronaria and characterization of novel molecules from its extract through high resolution-liquid chromatography mass spectrometry coupled to structural biochemistry approach

Lychnis coronaria, a perennial (herbaceous) belonging to Caryophyllaceae has been traditionally used for treating different complications. However, the free radical scavenging effect, anti-inflammatory activity and anticancer property of methanolic extract of this plant has not been addressed. Most importantly, the chemical constituents present in the extract of Lychnis coronaria responsible for its diverse activities have not been scrutinized till date. Here, we used a complex approach for exploring the above mentioned effects of Lychnis coronaria. We performed rigorous phytochemical screening followed by quantification of tannins, phenols, alkaloids, quinones and sterols from the extract. Moreover we employed in vitro DPPH, ABTS , FRAP assay, albumin denaturation inhibition experiment, MTT assay, high resolution liquid chromatography mass spectrometry for measurng the reactive oxygen species quenching, anti-inflammatory and anticancer strength of Lychnis coronaria and for identifying the possible bioactive molecules. We identified two novel molecules panaxynol (polyacetylenic alcohol) and norharman (9H-Pyrido [3, 4-B] indole) following rigorous analysis of the extract. Following this, the binding affinity of these molecules was estimated using human cyclooxygenase (COX)-2 enzyme as target. Among the constituents of Lychnis coronaria norharman manifested stronger binding towards COX-2 compared to panaxynol. Most importantly, norharman showed high stability in the groove of COX2 as confirmed by molecular dynamics simulation. Collectively, Lychnis coronaria manifested free radical neutralizing, inflammation soothing and anticancer effect in concentration dependent manner and thus may serve as a promising phytotherapeutic in future. Communicated by Ramaswamy H. Sarma

Characterization of tannin extracts derived from the bark of four tree species by HPLC and FTIR

The objective of this work was the extraction and quantification of tannins obtained from the bark of four tree species from the forests of Ixtlán de Juárez, Oaxaca, Mexico (Arbutus xalapensis Kunth., Prunus serotina Ehrh., Quercus crassifolia Humb. and Bonpl., and Quercus laurina Humb. and Bonpl.), In this extraction process, 90% ethanol was used as solvent at an extraction temperature of 70 °C for 2 h. The quantification of phenolic compounds, condensed tannins, and percentage of total extract, were obtained using the Stiasny number. The total extracts were 12.87, 16.38, 19.31, and 25.68% for P. serotina, Q. laurina, Q. crassifolia, and A. xalapensis, respectively. The condensed tannins were characterized by Fourier Transformed Infrared Spectroscopy (FTIR) where at 1626 cm−1 is observed an isomerization that implies a rotation between the C and B rings that corresponding at elongation of bonds in benzene structure of catechin. The purity of the extracted tannins was analyzed by High Performance Liquid Chromatography (HPLC). The effectiveness of the extraction process was proven; the importance of knowing the amount of tannins will promote the utilization of the bark of these species. The results of the condensed tannin content place Quercus crasifolia (10.29%) and Arbutus xalapensis (13.12%) as potential sources of tannins.

Tannin Content in Vitis Species Red Wines Quantified Using Three Analytical Methods.

Tannin content in red wines is positively correlated with astringency perception and wine grade; however, tannin quantification is one of the main challenges. In this study, tannin content was quantified using three analytical methods in commercial red wines from Vitis vinifera and interspecific cold-hardy hybrids including Marquette, Frontenac, and Petite pearl cultivars. Protein (PP) and methylcellulose precipitation (MCP) methods were compared to a HPLC-DAD method, which is based on the interaction between tannins and a hydrophobic surface (RPC). Frontenac wines were the poorest in tannins and Cabernet sauvignon wines were the richest regardless of the method used. In cold-hardy red wines, the tannin content was higher in Marquette with high alcohol content, which suggested that the tannins were extracted from seeds rather than skins. The high limit of quantification of the PP method and the presence of anthocyanin di-glucosides in cold-hardy wines were parameters suggesting that protein and methylcellulose precipitation methods were neither suitable nor reliable for the quantification of tannins in cold-hardy red wines. The tannin content quantified by RPC was positively correlated to tannin quantified by MCP, suggesting that the RPC method would be relevant for the quantification of tannins in red wines.

Comparative Study on Phytochemical Screening of Root and Bark with Leaf of Cardiospermum Halicacabum

Aim: The Present study was designed to compare the phytochemical screening of root and bark with leaf of Cardiospermum halicacabum. Materials and Methods: Samples were taken leaf (N=24) root and bark (N=24) based on the total sample size using clinical.com. The leaf, root and bark extract were collected. The phytochemicals were extracted by sequential extraction using three solvents methanol, ethanol and acetone. The quantification of flavonoids and phenols was performed by using Folin-Ciocalteu and quercetin as standard. Quantification of tannins was determined by using an insoluble polyvinyl-polypyrrolidone (PVPP) as standard. Results: Statistical analysis showed that methanol extract of root (0.49mg/ml) has highest phenolic content and acetone extract of root has highest tannin (0.64mg/ml) and flavonoid (1.18mg/ml) content when compared with leaf and bark. There appears to be a statistically significant difference in the mean of root when compared with leaf and bark (p<0.01, independent samples). Conclusion: In this study root appears to have better phytochemical and phenol content when compared with the content in leaf and bark.

Tannin quantification and chemical-energetic characterization of biomass residues of Bertholletia spp. and Lecythis’ spp. fruits

ABSTRACT Biomass residues originated from agroforestry systems has been demonstrating great usage potential for the pharmaceutical, textile and energy industries. In the Amazon region, the use of these materials can stimulate the sustainable economic development of extractive communities and agroforestry systems. However, for a better use of this material, it is necessary to quantify and qualify the chemical components present in these residues. Keeping that in mind, our research aimed to quantify the presence of condensed and hydrolysable tannins, and to chemically and energetically characterize the Bertholletia excelsa and Lecythis pisonis’ fruit residues, with the intent to assess the usage potential for this biomass resource in different industries. The studied material was collected from local extractives and merchants. Tannin quantification was performed through thin layer chromatography (TLC) method and the total values of extractives, lignin and holocellulose were also quantified. The energy potential was determined by the variables of apparent density, gravimetric yield, superior calorific power and charcoal’s immediate chemistry (the charcoal was produced at 450 ºC). The associations between variables were determined by a linear correlation. The TLC for condensed tannin was positive for B. excelsa. As for the TLC of hydrolysable tannins, the results were negative for both materials. The high levels of lignin and extractives in fruit’s residues of both species demonstrate the energy potential of this material. Our results may favor the use of B. excelsa’s residues for the production of polymers and adhesives, as well as other non-wood residues with high content of lignin, fixed carbon and calorific power in energetic products.

Determination of the tannin content extracted from the tara sheath (Caesalpinia spinosa) from the Portete neighborhood, Gonzanamá canton of the province of Loja

The tara (Caesalpinia spinosa)is a forest species native to South America, which presents a great potential in the world industry in the field of agro-export and agribusiness, for the use of its derivatives as tannins, which has anunmet demand in the market to be used in tanning and food industries; the present thesis project had the purpose of determining the amount of tannins present in the fruits collected in El Portete neighborhood, for this purpose, aqueous extracts were elaborated by maceration and heating, to which were make organoleptic (color and smell) and physical-chemical (p H and density) analyses, and, the quantification of tannins to each extract, considering two concentrations ofpotassium permanganate solution (0.025and 0.1 N). The results show that the organoleptic and physicochemical characteristics of the extracts obtained by heating and maceration did not present significant differences between them, except fo r the pH that ranges from moderately (3,7) to slightlyacid (4,3); as regards the quantification of tannins, there is a similarity between the values obtained with th e tw o extraction methods; however, there were differences between the value s re a ch e d w ith the two concentrations of potassium permanganate, obtaining the best yields (52,5 %) at the concentration of KMnO4at 0,1 N. Keywords:Potentiality, Quantification of tannins, Concentrations and Yield

Contents of tannins of cultivars of sorghum cultivated in Brazil, as determined by four quantification methods

High tannin content in sorghum grains is an undesirable characteristic for poultry and pig feeding and represents a challenge for breeding programs. On the other hand, moderate content of tannins in sorghum may be beneficial in human diets because they exert anti-cancer, anti-inflammatory and reduced carbohydrate uptake effects, among others. The aim of this study was to compare tannin contents of twenty sorghum cultivars available in Brazil, as well as to compare results obtained with four methods of tannin quantification: butanol/HCl, vanillin/HCl, BSA/FeCl3 and PVPP/Folin-Ciocalteu. The results obtained with butanol/HCl and vanillin/HCl were higher than with BSA/FeCl3 and PVPP/Folin-Ciocalteu. A known amount of purified quebracho tannin was used to test the validation of methods of tannin quantification and vanillin/HCl stood out for its high accuracy degree. The sampling used reveals wide genetic diversity regarding tannin contents. The expectation of predicting tannin contents on basis of grain color seems unfeasible.

UV–Vis Spectrometry for Quantitative Study of Tannin and Flavonoid Rich Dyes from Plant Sources

The UV–Vis absorbance method is modified as a faster and cheaper means for the study of extraction of natural dyes from plant sources. The method was validated using standard methods of analysis viz: egg albumin precipitation and aluminium colorimetric methods for quantification of tannins and flavonoids respectively. The solvents selected for extraction of dye from plant samples included; distilled water, methanol (5% v/v), ethanol (5% v/v), acidified water pH 3.5 (acidified with acetic acid), and sodium bicarbonate solution (0.1% w/w). Qualitative UV scans were conducted on dye extract of each plant obtained from previously dyed cotton fabrics and wavelengths at maxima for extract of each plant was recorded. In the validation step, data obtained from the modified UV absorbance method were compared to that obtained from standard methods. The modified and validated method was used for the study of extraction output of dye from selected dye-yielding plants using the selected solvents. Extracts from the selected plants registered absorbance maxima at wavelengths unique to each viz: A. coriaria (291 nm), V. paradoxa (294 nm), M.lucida (300 nm) and H. madagascariensis (428 nm) these were employed in setting the analytical instrument. Data obtained from the modified method closely correlated to that from egg albumin and aluminium colorimetric methods with R2 = 0.989 and 0.9258 respectively. The modified method registered minimal errors in the range of (0.3–8.7%) for the A. coriaria extracts and errors of (2.3–6.7%) for M. lucida extracts. The errors recorded from the standard methods were in the range of (2.1–7.9%) and (2.29–6.8%) for egg albumin precipitation and aluminium colorimetric methods respectively. Both the standard and the modified methods exhibited similar accuracy of analysis. Through the application of the modified method, sodium bicarbonate solution (0.1% w/w) was identified to give the best dye extraction output. This modified method gives a faster and cheaper means for quantitative analysis where data is required rapidly for decision making.

Miniaturized Spectrophotometric Method for Quantification of Tannins in Pomegranate (Punica granatum L.) Fruit Peel Dried Extracts

Background: Punica granatum L. (pomegranate) is one of the oldest edible fruits used for the treatment of countless diseases. The bioactivity of this fruit is mainly related to tannins, especially elagitannins. P.granatum fruit peel dried extracts (PPDEs) are widely marketed today and the lack of accessible quality control methods is a key to the variable and questionable quality of these products. Objective: To develop a miniaturized spectrophotometric method for quantification of tannins in commercial PPDEs. Method: Spectrophotometric method using Folin-Ciocalteu reagent was developed and validated for quantification of total polyphenols and polyphenols not adsorbed by hide powder in 17 samples of commercial PPDEs. Tannins content was calculated by the difference between total polyphenols and polyphenols not adsorbed by hide powder contents and expressed as pyrogallol. The UV/Visible microplate spectrophotometer was operated at 760 nm. Results: The developed miniaturized method was precise, accurate, selective and linear, and has no rotational or translational matrix effect. This method significantly reduced the cost of the analysis: 82.2% with reagents, 84.7% in electricity consumption, and 56.3% in the total analysis time. The wide variability in tannins contents in commercial PPDEs (<1% to 14.20%) shows the importance of quality control, ensuring the therapeutic effect and safety of the extracts. Conclusion: A miniaturized spectrophotometric method was successfully developed and fully validated, being appropriate for quantification of tannins of PPDEs in quality control routine analysis, as it showed to be a simple and cheaper method.

Methodology optimization for the analysis of phenolic compounds in chestnut (Castanea sativa Mill.).

Phenols are bioactive substances of great interest because of their involvement in plant physiology, their use in many industrial processes, and their impact on human health. This work aims to summarize the varied approaches to the phenolic analysis of chestnut (bark and wood of trunk and branches, leaves, catkins, burs, and fruit) and to collate the optimal conditions into an easy to follow and execute protocol. Phenolic compounds were extracted by solid-liquid extraction and separated by liquid-liquid extraction. Total phenols content was determined by Folin-Ciocalteu assay, condensed tannins by vanillin assay, and hydrolyzable tannins (gallotannins and ellagitannins) by high-performance liquid chromatography quantification of methyl gallate and ellagic acid following acid methanolysis. The lowest temperature for conservation (-80 ℃), lyophilization, and milling (liquid N2) were the most effective pretreatments for samples. For quantification of tannins, the use of water clearly reduced the sensitivity of the analysis of condensed tannins, whilst the more efficient degradation capacity of sulfuric acid improved the methanolysis of hydrolyzable tannins. These findings were validated using a range of chestnut tissues, and thus confirm the utility and effectiveness of this easy to implement, cost-effective, and efficient protocol.

Characterization and quantification of tannins, flavonols, anthocyanins and matrix-bound polyphenols from jaboticaba fruit peel: A comparison between Myrciaria trunciflora and M. jaboticaba

Jaboticaba, a polyphenol-rich fruit, has numerous nutraceutical properties. Studies on jaboticaba composition have focused on solvent-extractable polyphenol compounds, whereas the matrix-bound polyphenols that can be delivered during digestion have been neglected. This study aimed to characterize the polyphenol profiling of matrix-bound and free phenolic compounds in Jaboticaba peel powder (JPP) using two fruit species, Myrciaria jaboticaba (JPP-MJ) and M. trunciflora (JPP-MT). An HPLC-DAD-MS/MS method was developed and validated to analyze JPP polyphenols. The total content of free phenolic compounds was 2.4 times higher in JPP-MT than in JPP-MJ and the profile of free polyphenols differed between the two species. JPP-MT had 60.3% tannins and 33% anthocyanins, whereas JPP-MJ had 34.7% and 50.7%, respectively. The content of matrix-bound phenolic compounds was higher in JPP-MJ than in JPP-MT (15.3% vs. 4.3% of total phenolic compounds). Besides cyanidin 3-glucoside, three other compounds were identified as major: a tetragalloylglucose isomer for both species, trigalloylglucose for JPP-MT, and delphinidin-3-glucoside for JPP-MT. Phenolic profiling of JPP revealed new compounds that may contribute to health benefits after JPP consumption. Despite differences in the phenolic profile between the jaboticaba species, both JPPs are rich sources of polyphenols and could be used for promoting health benefits.

Quantification of Tannins from Curupay Bark

ABSTRACT This study aimed to assess the influence of time and sodium sulfite concentration on tannin yield in Curupay bark and propose better extraction conditions. The extractions were performed using 100 g of dried bark and 1500 mL of water at 70 °C, with extraction times of 2, 3, and 4 h and amount of extractor salt at the concentrations of 0, 1, 2, 3, and 4%. The following parameters were evaluated: content of total solids, total yield of solids, Stiasny index, and gravimetric yield of condensed tannins and non-tannins. Results show that the higher the concentration of sodium sulfite, the higher the content and yield of solids, as well as the higher the extraction of non-tannin compounds. Three-hour extraction time at 3.0% sodium sulfite concentration was the best condition to extract tannins from Curupay bark, under which a condensed tannin yield of 20.46% was obtained.

Evaluation of the Folin-Ciocalteu Method and Quantification of Total Tannins in Stem Barks and Pods from Libidibia ferrea (Mart. ex Tul) L. P. Queiroz

ABSTRACT The stem bark and pods of “juca” are widely used in Brazilian folk medicine and, despite their therapeutic relevancies, there are insufficient reports about their quality control. Both herbal drugs from the species are rich in tannins, which showed a lot of biological and economic interest. Thus, the purpose of this study was to evaluate the method for quantification of tannins in the stem bark and pods from L. ferrea. The performance of the method was tested against the influence of the most relevant parameters (reaction time; amount of drug; type and amount of precipitationagents; and, concentration of the reagents). The procedure was validated according to the literature. After optimization, the experimental conditions were established as: 30 min of reaction; λ of 760 nm; 1.0 mL of Folin-Ciocalteu; 29% of Na2CO3 (w/v); and, skin powder as complexion agent. The method validation showed that the instrumental response was linear and sensitive to the investigated analyte. The method presented the necessary precision (RSD < 2.19%) and accuracy (96.84%-109.13%), as recommended by the official codex. Moreover, the deliberate introduction of experimental variations showed the stability of the procedure error against external sources (RSD < 2.42%). In conclusion, the data demonstrated the suitability of the method and can be used as anappropriate analytical tool for quality control of herbal drugs from L. ferrea.

Acute and chronic toxicity and antimicrobial activity of the extract of Stryphnodendron adstringens (Mart.) Coville

ABSTRACT: This study evaluated the antimicrobial activity and acute or chronic toxicity of the extract of Stryphnodendron adstringens. The stem bark dry extract was obtained by static maceration with ethanol. Quantification of tannins was performed by the Folin-Denis method, which indicated a total tannin content of 32.7%. The antimicrobial activity of the dry extract of S. adstringens was evaluated by agar-based disk diffusion assay with Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) in the concentration of 200, 400 and 600μL/mL. The results indicated that 600μL/mL inhibited microbial growth, i.e. had antimicrobial activity against these species. Acute and chronic toxic effects of S. adstringens was evaluated in Wistar rats treated with 200, 400, 600 and 800mg/kg of extract, administrated by gavage. Liver degeneration was observed in the group of rats receiving 800mg/kg in chronic exposure, what may indicate some degree of toxicity at this concentration. However, no systemic toxicity was observed at lower doses. Considering the broad use of S. adstringens as a phytotherapeutic agent for various human and animal diseases and the livertoxicity observed at high concentrations, attention should be paid to the possible adverse effect of using the extract from this plant at high concentration.

Quantification of tannins and related polyphenols in commercial products of tormentil (Potentilla tormentilla).

Potentilla tormentilla has many biological and pharmacological properties and can be used as an ingredient of some herbal medicines or beverages. The aim of this study was to evaluate the content of individual polyphenols, especially condensed and hydrolysable tannins in commercially available tormentil rhizomes and tinctures using chromatographic methods. A quantitative analysis (HPLC-PDA) was preceded by qualitative studies (UPLC-qTOF-MS/MS) and the isolation (CC) of the major tannin compounds. The tested plant material is characterised by a high content of tannins and related polyphenols, i.e. in rhizomes even at the level above 20% and in tinctures above 2%. The main components of tormentil rhizomes are procyanidin B3 (mean ~ 3.6%), procyanidin C2 (mean ~ 2.8%), agrimoniin (mean ~ 2.5%), 3-O-galloylquinic acid (mean ~ 1.7%), catechin (mean ~ 1.6%), other flavan-3-ol oligomers (mean ~ 0.5-1.1) and laevigatins (mean ~ 0.2-0.6%). Free ellagic acid and glycosides of ellagic and methylellagic acids are secondary components. Underground parts of tormentil are a source of oligomeric proanthocyanidins and ellagitannins, but in smaller quantity of gallotannins. Monogalloylquinic acids are new identified compounds, which had not been described in Potentilla tormentilla before we started our research. In the analysed tormentil tinctures agrimoniin concentration is lower in relation to other tannins.

Robust Ultraviolet-Visible (UV-Vis) Partial Least-Squares (PLS) Models for Tannin Quantification in Red Wine.

The validation of ultraviolet-visible (UV-vis) spectroscopy combined with partial least-squares (PLS) regression to quantify red wine tannins is reported. The methylcellulose precipitable (MCP) tannin assay and the bovine serum albumin (BSA) tannin assay were used as reference methods. To take the high variability of wine tannins into account when the calibration models were built, a diverse data set was collected from samples of South African red wines that consisted of 18 different cultivars, from regions spanning the wine grape-growing areas of South Africa with their various sites, climates, and soils, ranging in vintage from 2000 to 2012. A total of 240 wine samples were analyzed, and these were divided into a calibration set (n = 120) and a validation set (n = 120) to evaluate the predictive ability of the models. To test the robustness of the PLS calibration models, the predictive ability of the classifying variables cultivar, vintage year, and experimental versus commercial wines was also tested. In general, the statistics obtained when BSA was used as a reference method were slightly better than those obtained with MCP. Despite this, the MCP tannin assay should also be considered as a valid reference method for developing PLS calibrations. The best calibration statistics for the prediction of new samples were coefficient of correlation (R2val) = 0.89, root mean standard error of prediction (RMSEP) = 0.16, and residual predictive deviation (RPD) = 3.49 for MCP and R2val = 0.93, RMSEP = 0.08, and RPD = 4.07 for BSA, when only the UV region (260-310 nm) was selected, which also led to a faster analysis time. In addition, a difference in the results obtained when the predictive ability of the classifying variables vintage, cultivar, or commercial versus experimental wines was studied suggests that tannin composition is highly affected by many factors. This study also discusses the correlations in tannin values between the methylcellulose and protein precipitation methods.

Tannin quantification in industrial waste of tobacco production and selection of optimal extraction parameters

Optimum technological parameters of extraction for the sum of tannins from tobacco dust (Nicotiana tabacum L) waste of tobacco production were established. The basic process parameters to obtain the highest yield of tannins from raw materials such as selection of optimum extractant ratio of raw material to solvent, extraction time, temperature and mode, selection of the optimal precipitator for tannins and the optimal ratio of extract to precipitator. A process technological scheme of tannins extraction from tobacco dust waste was suggested.

In vitro ovicidal and larvicidal activity of methanolic leaf extract of Manihot esculenta (cassava) on susceptible and resistant strains of Trichostrongylus colubriformis

This study aimed to represent the first report of the ovicidal and larvicidal activity of the methanolic leaf extract of Manihot esculenta (cassava) against eggs and larvae of susceptible and resistant strains of Trichostrongylus colubriformis. As well as, to determine the total tannin compounds, antioxidant activity and toxicity of the extract. The egg hatch test was used to evaluate ovicidal activity against unembryonated eggs, whereas larval feeding inhibition assay and MTT-formazan assay were used to evaluate larvicidal activity against first (L1) and infective (L3) larvae, respectively. The results showed no significant differences were detected between the sensitivities of susceptible and resistant strains of T. colubriformis to the extract. Eggs, L1 and L3 were significantly affected (P<0.001) compared with negative control, and L1 were more sensitive than the eggs and L3. The total tannin compounds were investigated using tannin quantification assay and determined by 254.44TAE/mg. The antioxidant activity was evaluated using the DPPH radical scavenging assay and the median inhibition concentration (IC50) was determined by 2.638mg/ml. Acute oral toxicity at dose of 5000mg/kg, and sub-chronic oral toxicity at 500 and 1000mg/kg of the extract were observed in male and female Sprague-Dawley (SD) rats. The acute oral toxicity revealed that the median lethal dose (LD50) of methanolic extract of cassava leaves on SD rats was greater than 5000mg/kg, whereas the sub-chronic oral toxicity did not show observed adverse effects at 500 and 1000mg/kg per day for 28days. In conclusion, the methanolic extract of cassava leaves has direct ovicidal and larvicidal activity against T. colubriformis strains with a safety margin for animals, and it may be potentially utilized as a source of natural antioxidants.