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Phytochemical Methods
Abstract
The macromolecules of plants are distinguished from all other constituents by their high molecular weight. This may vary from 10000 to over 1000000, whereas in other plant metabolites the molecular weight is rarely above 1000. Chemically, macromolecules consist of long chains of small structural units or ‘building blocks’, linked covalently in a number of different ways. Chemical characterization in the first instance therefore depends on identifying these smaller units. Proteins, for example, are long chains of amino acids (up to twenty different ones) joined together through peptide (—CO—NH—) links. Polysaccharides are similarly derived from the union of simple sugar units, such as glucose, joined through ether (—O—) links. The nucleic acids, by contrast, are more complex and have three types of structural unit: purine and pyrimidine bases, pentose sugars and phosphate groups. The three main classes of macromolecules found in plants are thus proteins, polysaccharides and nucleic acids. However, mixed polymers are also known. such as the glycoproteins, which contain both sugars and amino acids in covalent linkage.
... The precipitate was removed by filtration process by using Buchner funnel then the filtrate was concentrated under vacuum by using f reeze drier to get the crude . By the same method cold ethanolic extract was prepared but by using ethanol as a solvent ( Harborne ,1984 ;Bruneton , 1995). ...
... Presences of phenolic compounds The differences of Rf values of four phenolic compounds belong to variance in many factors such as their polarity , molecular weight and stereochemistry . Also using of TLC as an analytical technique for separation of phytochemical active compounds including phenols , ensures importance and advantage of this chromatographic method in isolation and purification of these compounds because it has many features such as accuracy and good separation (Harborne ,1984). ...
Diabetes mellitus is a multifactorial disorder characterized by hyperglycemia resulting from increased hepatic glucose production. The current study was carried out to investigate the hypoglycemic action of synergistic interaction of phenolic compounds isolated from Iraqi Phoenix dactylifera (Breim) leaflets. Infra-red and gas chromatography-mass spectroscopies were applied for identification of the phenolic compounds which are represented by alpha-tocopherol , 2-hydroxy-5-methylisophthalaldehyde , 3-hydroxy-tyrosine and 4-n-propylresorcinol. The phenolic compounds recorded a significant decreasing after 4 hours (280.1 mg / 100 ml) and highly significant decreasing at 6 hours (238.3 mg/ 100 ml) and 24 hours (134.5.mg / 100 ml) after oral administration in hyperglycemic rabbits. The phenolic compounds had no toxic effect on red blood cells since no hemolysis was found towards these cells, this means phenols Isolated can be used safely to treat diabetes mellitus disease instead of insulin drug but this work demands further clinical and pharmaceutical studies .
... Phytochemical screening: For the preliminary phytochemical investigation of prickle extracts were done according to methods described in Harborne (1973) [7] and Kokate (1997) [8]. The different extracts obtained by successive solvent extraction were tested separately for the presence of various phytoconstituents, viz. ...
... Phytochemical screening: For the preliminary phytochemical investigation of prickle extracts were done according to methods described in Harborne (1973) [7] and Kokate (1997) [8]. The different extracts obtained by successive solvent extraction were tested separately for the presence of various phytoconstituents, viz. ...
... The study would provide valuable information regarding the medicinal and pharmaceutical importance of E. debile for the researchers. subjected to Phytochemical screening by following the procedures as described by Trease and Evans [22] and Harborne [23]. ...
... To this, the addition of concentrated sulphuric acid (1 mL) resulted in to the formation of brown ring at the interface of two layers, which indicated the deoxysugar characteristic of cardenolides. Estimation a. Tannins: Tannins were extracted in 20% methanol using method as described by Harborne [23] and estimated by following the method of Makkar [24] as modified by Fagbemi et al. [25]. The tannins content were calculated as g/100 g dry weight from standard curve of tannic acid (R 2 =0.9944). ...
Biochemical, phytochemical and antioxidant composition of Equistem debile R. stem was determined. The E. debile extract showed the presence of reducing and non reducing sugars, total sugars, free amino acids, water soluble proteins and salt soluble protein. Phytochemical screening confirmed the presence of tannins, saponins, flavonoids, terpenoids, cardiac glycosides and total phenolic acid contents. Trolox equivalent total antioxidant activity of methanolic extract determined by DPPH radical scavenging assay was found to be lower than that determined by Phosphomolybdenum assay while the ascorbic acid equivalent total antioxidant activity determined by DPPH radical scavenging assay was found to be higher than that determined by Phosphomolybdenum assay. High value of ABTS+ radical scavenging activity and low values of DPPH and hydroxyl radical scavenging activity and reducing power have been observed in E. debile extract as compared to Trolox and ascorbic acid. The data provides useful information regarding the nutritional and medicinal importance of E. debile and suggest that this plant possesses good antioxidant activity.
... The leaves were ground using an electric grinder to obtain a fine, dry powder and then placed in dry, airtight bottles in the refrigerator at a temperature of 4°C until use. The cold aqueous extract was prepared according to the method of [ 8 ] , where 10 g of dry powder were taken and mixed with 200 ml of distilled water using an electric mixer and left for 24 hours at room temperature, after which the mixture was filtered using several layers of medical gauze to get rid of suspended particles then, centrifugation was carried out at a speed of 3000 rpm for ten minutes. After that, the extract was filtered using filter paper to obtain a clear extract. ...
This study examined the efficiency of cold aqueous extract of geranium, neem, and peppermint plants in mortality in different stages of the peach fruit fly, Bactrocera zonata. The results indicated that each of the three extracts had an effect on the third larval instar. The results showed that neem extract had the highest effect on the third larval instar. The cumulative corrected mortality rate after 72 hours reached 67.3%, while the corrected mortality rate of larvae due to the effect of peppermint extract was 58.6%. As for the geranium extract had the least effect among the three extracts on the larval stage, as the corrected cumulative mortality rate reached 52%. As for the effect of the aqueous extracts of the three plants on the stage of pupation, the neem extract was the most effective among the three extracts, as the highest rate of non-emergence of adults at a concentration of 7% was 30%. As for peppermint extract, the mortality rate was 10%, while for geranium extract, the mortality rate was 6%, There was no effect on the pupal stage at the concentration of 3%. These compounds also showed a clear effect in repelling the adult stage, as the geranium extract had a significant impact in repelling insects, as the insect repellency rate reached 72.16%. As for the neem plant extract, the insect repellent rate was 50.03%, while the peppermint plant extract showed a 62% repellent rate for B. zonata adults.
... 20 Preliminary phytochemical analysis Ethanol extraction of S. torvum fruit yielded 3.6% from 1000 g of dried fruit. Following conventional protocols, preliminary phytochemical analyses of the ethanol extract of matured fruits of Solanum torvum were conducted to identify the main phytochemical elements, including alkaloids, proteins, carbohydrates, tannins, sterols, triterpenoids, saponins, and flavonoids [21][22] Acute toxicity studies Acute toxicity studies were conducted on male and female Wistar rats following slightly modified OECD-423 guidelines. The animals were divided into five groups: males and females. ...
Solanum torvum Swartz , a member of the Solanaceae family, is a significant medicinal plant that is found all over the world and is used in traditional medical systems to treat conditions like diabetes, high blood pressure, tooth decay, and reproductive issues. The study aimed to evaluate the anticonvulsant and neuroprotective effects of Solanum torvum fruit extract (STF) in rodent models and to assess its acute and subacute toxicity. Acute toxicity was assessed in Wistar rats at doses up to 4000 mg/kg, while subacute toxicity was evaluated over 30 days at doses of 125, 250, and 500 mg/kg. The anticonvulsant activity was evaluated using Maximal Electroshock (MES) and Pentylenetetrazole (PTZ)-induced seizure models at 200 and 400 mg/kg. The neurotransmitter levels (acetylcholine and dopamine) and oxidative stress markers (superoxide dismutase [SOD], catalase [CAT], and malondialdehyde [MDA]) levels in the brain were measured to explore their neuroprotective effects. The phytochemical analysis revealed the presence of various bioactive compounds. Acute and subacute toxicity studies showed no adverse effects on body or organ weight, haematological parameters, or organ health, confirming safety. In the MES and PTZ models, STF demonstrated dose-dependent anticonvulsant effects, significantly reducing hind limb tonic extension (HLTE), hind limb tonic flexion (HLTF), jerking duration, and increased survival, achieving 100% protection at 400 mg/kg in the PTZ model. STF reduced Acetylcholine (Ach) levels and partially restored dopamine levels, counteracting neurochemical imbalances in the convulsion's pathophysiology. In addition, STF increased SOD and CAT levels and reduced MDA levels, proving its antioxidant potential. STF is observed as safe and exhibits significant anticonvulsant and neuroprotective properties, particularly at 400 mg/kg. Its ability to modulate neurotransmitters and reduce oxidative stress suggests its potential as an adjunctive therapy for convulsive disorders, warranting further exploration of its mechanisms and therapeutic applications.
... The detector is configured specifically for the detection of various flavonoids quercetin was detected at 370 nm and kaempferol at 367 nm, while luteolin and apigenin exhibited absorption maxima at 350 nm and 340 nm, respectively. The initial analysis of the sample extract involved the setup of a UV detector based on recommendations from reference book [10] for details on the spectral maximum of flavonols and flavones. The high-performance liquid chromatography conditions for M. hexandra stembark extract was fine-tuned based on literature data. ...
... The residue was the alkaloid, which was dried and weighed (Harborne, 1973). Total phenol content of millet flour sample was determined following Singleton et al. (1999). ...
Finger millet is a rich source of protein, dietary fibre, calcium, phosphorous, iron, zinc, other minerals
and vitamins along with some anti-nutrients. The anti-nutrients present in finger millet grains bind with
certain minerals such as iron, calcium, zinc to form soluble as well as insoluble complex compounds.
Thus, these anti-nutrients decrease the bioavailability of these minerals. Malting is a pretreatment that
helps in increasing the bioavailability of nutrients through inducing hydrolytic activity. Thus to optimize
the availability, digestibility and rate of absorption of all the nutrients present in the grains, malting can
be an efficient process. The aim of the present study was to determine the effect of malting on nutrient
and anti-nutrient composition of finger millet flour. A local variety of brown finger millet was selected to
evaluate the nutritional changes during the malting process. The nutrient profile of the malted finger
millet flour showed significant increase in protein (8.64g vs. 7.52g), carbohydrate (68.84g vs. 67.12g),
calcium (372 mg vs. 356 mg), iron (4.42 mg vs. 4.12 mg), magnesium (143.2 mg vs. 137.1 mg) and
zinc (2.54 mg vs. 2.47 mg) content and a slight decrease infat (1.9g vs. 1.3g), dietary fibre (10.71g vs.
9.98g), sodium (3.23 mg vs. 4.67 mg)and phosphorous(216 mg vs. 231 mg) content as compared to
raw finger millet flour per 100 g. The total ash content decreased insignificantly, while the potassium
content increased non-significantly during malting process. Additionally, the total phenol and alkaloid
contents increased during malting. Significant reductions in phytate, tannin and saponin content were
observed in malted finger millet flour as compared to raw finger millet flour.
... A cold-water extract of the mint plant was prepared according to [6], modified from [7]. (10 gm) of mint plant powder mixed with (200 ml) of cold distilled water on a magnetic stirrer for 15 minutes, then the solution left for 24 hours (to get a better extraction) after tightly covering it to avoid impurities entering. ...
This study was aimed at evaluating whether Syzygium Aromaticum extract, based on plant materials immersed in cold or alcohol, was effective at any life stage of Arboridia Hussaini Ghauri. The outcome of the study collected the date that showed the effect of cold water extract with three concentrations (0.50, 0.75, and 1.00) mg/ml on the life stages of the insect which differs in degree, as concentrations with of 1.00 mg / ml yielded the highest rate of mortality at 74.1% in the nymphs and The alcoholic extract severs mortality of nymphs and adults at were (81.5% and 70.4%) after 5 days with the corresponding mortality rates being the same for the particular concentration of this toxicant. The outcomes of the analysis indicated that a high concentration of the extract had a promise of a high mortality rate.
... The samples were then allowed to cool before proceeding with their qualitative phytochemical analysis. Harborne (1973), Trease and Evans (1989) and Sofowora (1993) was used to detect the phytochemical constituents of leaves, such as the leaves of T. daniellii and M. paradisiaca. Qualitative phytochemical screening was carried out on these leaves to evaluate them for the presence of saponins, polyuronides, phenolic compounds, reducing sugar, cyanogenic glycosides, alkaloids, flavonoids, triterpenes, phytosterols and anthracenosides (Ciulei, 1981). ...
Leaves from the plants of Thaumatococcus daniellii and Musa paradisiaca are used for wrapping foods in Ghana. In this study, the types of phytochemicals present in the two leaves were qualitatively determined. Standard qualitative phytochemical tests by Trease and Evans, Sofowora and Harborne were employed in screening for ten selected phytochemicals. The following phytochemicals were present in T. daniellii leaf extracts; saponins, polyuronides, phenolic compounds, reducing sugars, alkaloids, flavonoids, cyanogenic glycosides, phytosterols and anthracenosides. However, triterpenes were absent. M. paradisiaca also showed the presence of saponins, phenolic compounds, cyanogenic glycosides, polyuronides, reducing sugar, flavonoids and phytosterols. However, alkaloids, triterpenes and anthracenosides were absent. From the results obtained, T. daniellii contained more classes of healthful phytochemicals than that of M. paradisiaca. Since, the leaves of both T. daniellii and M. paradisiaca contribute more than just a means of packaging to these local delicacies, their continued use should be highly encouraged and widely promoted.
... The fruiting bodies of four termite mushrooms were collected from Chitwan National Park, Nepal. They were identi ed based on their morpho-taxonomic characteristics concerning the relevant literature [17][18][19] [20]. The methanol extract of termite mushrooms was tested for the qualitative screening of secondary metabolites like alkaloid, volatile oil, avonoid, steroid, saponin, tannin, emodin, polyurenoid, phenolic compound, amino acid, reducing sugar and anthocyanin. ...
Objective
Diabetes mellitus is characterized by the increase in glucose levels in the blood and serum because of a change in carbohydrates, fat and protein metabolism. Termite mushrooms are consumed for high nutrition and traditional medicine. The methanol extract of the four termite mushrooms; Termitomyces albuminosus, T. eurhizus, T. robustus and T. striatus f. ochraceus was tested for inhibition of alpha-amylase by the starch-iodine method, and effect on blood glucose level and body weight in alloxan-induced diabetic mice.
Results
The secondary metabolites in the methanol extract of termite mushrooms were screened. Alkaloid, flavonoid, polyose, steroid, glycoside, terpenoid, fatty acid and emodin were found in all the termite mushrooms. The extract of T. straitus f. ochraceus has shown a strong inhibition against porcine pancreatic alpha-amylase by 61.4% at 200 µg/ml concentration. The extracts of all termite mushrooms at 500 mg/kg showed a significant reduction in the blood glucose level after 15 days of administration. Extract of termite mushrooms in two doses (250 and 500 mg/kg) intraperitoneally administered to the alloxan-induced diabetic mice indicated a significant enhancement in their body weight as compared to diabetic mice. Hence, termite mushrooms can be used as a substitute supplement for the management of diabetes mellitus.
... Preliminary phytochemical analysis of Tamarindus indica.is done as per method described by Wagner et al. (1908), Harborne (1988) and Eike and Anne (2006). HPTLC profiling is done by using CAMAG HPTLC System with WIN CATS software. ...
From last two to three decades, we are facing lots of infectious diseases
and pandemic. It reveals that we are forgetting our traditional food,
medicine and practices. People globally were consuming their food as
per climates, seasons and landscapes, but due to urbanization adopting junk and
unsuitable foods. The metallic life also leading to poor immunity and therefore,
the documentation of such traditional plants having food and medicinal values
associated with a particular boundary is the need of hour. Now also need the
culture of – “Let food be the medicine and medicine be the food”. In this aspect,
the published book chapters in the book entitled “Medico-Biowealth of India”
Vol VI, will provide a baseline data for future advance research works in
mitigating the contemporary health problems. I wish the book will be helpful to
the researchers, academicians and intellectuals.
... Chromatograms were developed in a system of n-butanol: acetic acid: water (50:2:48) by ascending chromatography on silica gel TLC (F254) plates using various phenolics as reference compounds. Phenolic principles were detected using Rf-values, ferric chloride-ferric cyanide reagent and UV light (+NH 3 vapours) (Harborne, 1973 ...
The study of alleopathic potential of Seriphidium leucotrichum (Krasch. ex Ladyg.) K. Bremer & Humphries ex Y.R. Ling, on seed germination and early seedling growth of four test plant species including Triticum aestivum (wheat), Zea mays (maize), Brassica compestris (mustard) and Pennisetum americanum (millet), was conducted under laboratory conditions. Aqueous extract of the air-dried plants of S. leucotrichum at different concentrations i.e. (25, 50, 75 and 100% of stock solution) inhibited germination of the test species in the order: Brassica compestris> Zea mays> Triticum aestivum> Pennisetum americanum. Root and shoot growth of the test species was also reduced by the extracts in the order: Brassica compestris> Zea mays> Triticum aestivum > Pennisetum americanum. It was observed that the soil application of the aqueous extract had considerable delaying effect on Triticum aestivum growth while shoot spray or root dip treatment had no such effects. Decaying shoot of S. leucotrichum in sandy-loam at 5, 10 and 20g/400g soil caused considerable inhibition of germination and seedling growth of Pennisetum americanum at high concentration (20 g/400g soil). Bioassay of the ether extract of S. leucotrichum exhibited three zones of inhibition at Rf values 0.7-0.8, 0.8-0.9 and 0.9-1.0 while a promoter was detected between Rf value 0.4-0.5. Paper Chromatography was employed for the identification of phenolic constituents and as a result, caffeic acid, p-coumaric acid and gallic acid were identified while one constituent remained unknown. In this connection, the possible role of the phenolic compounds through their allelopathic effect in driving the structure of natural communities is discussed.
... Quercus rubra fruit casings of used were purchased from local markets in the city of Najaf in October of 2019, and have been diagnosed by taxonomists in the Botanical Laboratory for Graduate Studies at the College of Science -University of Kufa. After that it was cleaned and isolated from foreign materials, crushed by an electric mill, then the powder was collected in nylon bags and kept in the laboratory at room temperature until use (Harborne, 1984). Alcoholic extracts 30 grams of powder was taken and 300 ml of of alcohol solvent (Methanol 99% and n-Hexan 96%) was added in a 500 ml lask. ...
This study was designed to investigate the ability of methanol and n-hexane extracts of fruit casings of Quercus rubra against pathogenic bacteria isolated from patients with burn infection. Three concentrations of methanol and n-Hexane extracts of Quercus rubra were done (100, 200 and 400) mg/ml. Thirty eight samples were collected from Medical Burn Center in Najaf Gover-norate. Males and females in different age groups. The results indicated that there were 42% pseudomonasaeroginosa, 22% Staphylococcusaureus, 18% Klebsiellapneumonia, 8% E. coli, 6% Staphylococcusepedermsis and 4% pro-teusmiribilis. Seventeen antibiotics were selected in antibiotics sensitivity test. Imipenem 10mg was the best antimicrobial effect. The results showed that all three concentrations had good antibacterial effect. It has also been shown that methanol solvent is more effective on bacteria than n-hexane solvent. Also, the results showed that P. aeroginosa and S. auereu has the highest rate of inhibition diameter with methanol solvent (38.123 ± 0.07 and 35.133 ± 0.092) mm respectively. While K. pnemeumonia was the highest rate of inhibition diameter by n-hexane solvent 32.4 ± 0.07mm. Methanol and n-hexane extracts of Quercus rubra can be regarded as a potential antibacterial against aerobic pathogenic bacteria isolated from patients with burn infection. Therefore , Fruit casings of Quercus rubra may be considered as a raw material for the manufacture of ointment for treatment of burns infections.
... Quantitative phytochemical analyses for alkaloids, tannins, cyanogenic glycosides, oxalate, saponins, flavonoids, phenols and phytates in the processed seed sample were determined by the methods described by Harborne (1973) and Trease and Evans (1989). ...
The chemical composition of the ground seeds of Picralima nitida were determined using standard methods. The results of the phytochemical analyses indicated the presence of alkaloids (5.33±0.57%), tannins (9.60±0.05%), cyanogenic glycosides (3.39±0.03%), oxalates (4.36±0.02%), saponins (13.50±0.50%), flavonoids (5.50±0.40%), phenols (1.79±0.03%) and phytates (0.17±0.004%). The moisture content of the seed was 10.67±0.34%, ash 3.67±0.34%, protein 3.50±0.18%, crude fiber 8.78±0.60% fat 3.49±0.10% and carbohydrate 69.90±078%. The seeds contained vital essential amino acids especially leucine (11.83%), phenylalanine (9.21%) and tyrosine (6.08%) and non-essential amino acids. The ground seed contained more unsaturated fatty acids (78.87%) than saturated fatty acids (20.13%). There were appreciable presence of macro and micro-elements with iron (172.40±0.70), zinc (55.70±0.30) and manganese (38.20±0.20) mg/kg being high. The ground seeds had vitamins A 123.40±0.30 and E 3285.70±0.05 µg/100 respectively. The study revealed that Picralima nitida seeds contain important phytochemicals, amino acids, fatty acids, vitamins and minerals which, if used, will not only offer medicinal benefits to its users but could also serve as a good source of nutrients.
... In addition, the lead acetate test was also employed for phenol assessment. The methods proposed by Harborne [30] included the addition of 10% ...
Objective. This study was designed to determine the antioxidant activity of the extracts of beetroot by using beetroot energy drink and flavored milk (products).
Material & Methods. This Experimental trial was conducted at Jinnah University for Women, Pakistan, under the approval of local Institutional review board number JUW/ DFST / RCB-010/2020. All the materials such as beetroot, carrot, cucumber, and lemon were obtained commercially from which two products were formulated: beetroot energy drink (sample1) and flavored milk (sample 2). These formulated products were evaluated for quality analysis (pH and Brix), phytochemical screening using the Keller-Killani test, Salkowski’s test, Alkaline Reagent Test, Lead Acetate Test, and Ferric Chloride Test, protein test, quantitative test of phenol, antioxidant activity, sensory analysis, and shelf-life study. Paired t-test was applied to detect significant differences between two samples.
Results. The phytochemical analysis revealed that cardiac glycosides, phytosterol, flavonoids, and Terpenoids were found in both Energy Booster Drink (EBD) and Flavored Milk (FM) except Phenolic compounds that were found only in EBD. The antioxidant capacity of beetroot juice was far greater than FM. The statistical sensorial analysis of FM and EBD reported a significant mean difference between most of the groups with p<0.0001.
Conclusion. This study concludes that energy drinks having beetroot indicated higher anti-oxidant capacity than flavored milk. The nutraceutical products (Energy booster drink and Flavored milk) containing beetroot are enriched with optimum quantities of proteins and fats, and low carbohydrates at a stable pH with adequate total energy content.
... Phytochemical screening was carried out on the ethanolic extract of V. amygdalina following the methods previously described by Harborne [25] and Trease and Evans [26]. The ethanolic extract of V. amygdalina was screened for alkaloid, saponin, tannin, flavonoid, and terpenoid. ...
The adsorption and corrosion inhibition performances of the phytoconstituents from Vernonia amygdalina (bitter leaf): alkaloid (AEVA), flavonoid (FEVA), saponin (SEVA), tannin (TAVA), and terpenoid extracts (TEVA) on mild steel in acidic media were investigated by gravimetry, gasometry, potentiodynamic polarization, FT-IR, and quantum chemical calculation. The experimental results revealed excellent inhibition properties for the extracts, with FEVA and SEVA possessing the highest corrosion inhibition efficiencies. The calculated values of activation energy were all positive and decreased in the inhibited systems when compared with the uninhibited system, with an exception for TAVA. Potentiodynamic polarization curves indicated that all the studied inhibitors were mixed-type with greater cathodic reaction inhibition tendency, while FT-IR revealed that the mechanism of adsorption of the interfacial species on the mild steel surface was by chemisorption, and quantum chemical calculation was useful in highlighting the contributions of the phytochemicals to the corrosion process.
Graphic Abstract
... Phytochemical screening of the obtained extracts was carried out using standard qualitative methods as described by various researchers [9][10][11] . The extracts were analyzed for the presence of alkaloids by Mayer and Dragendorffs tests, carbohydrates by Wagner's, Hager's, Molisch and Fehling's tests, glycosides by Borntrager's and Legal's tests, tannins and phenolic compounds by Ferric chloride and lead acetate tests, proteins and amino acids by Million's and Biuret tests, flavonoids by Shinoda and Alkaline reagent tests, presence of steroids and triterpinoids by Liebermann-Burchard's and Salkowskis tests. ...
The present study was aimed to explore the potential of Musa acuminata var G9 leaves (banana leaves, BL) as a wound healing agent and to formulate a topical delivery system for delivering the same. BL was collected, authenticated and the leaf components were extracted by microwave-assisted extraction (MAE) using three individual solvents. Phytochemical screening of three extracts led to the selection of aqueous extract which was further subjected to by GCMS analysis. MTT assay of aqueous BL extract exhibited cytocompatibility even at higher concentrations (150 mg/mL showed 85.45%). Antibacterial studies using agar disc diffusion method revealed more susceptibility of S. aureus in comparison to E. coli for the aqueous extract. For topical application, four creams (F1-F4) were developed and characterised. Further, scratch wound healing potential of 25 mg/mL and 50 mg/mL of aqueous extract in F4 formulation was compared with marketed and control formulations using L929 cell lines which exhibited better cell migration of 25 mg/mL extract in 24 h with wound area of 44.46 ± 3.34 % compared to marketed 32.41 ± 1.63 % and control 9.38 ± 1.76 %. Thus the study indicates the potential of aqueous BL extract as a wound healing agent which can be further explored.
... Filtrate was discarded and the residue is the alkaloid was dried and weighed. 29 ...
Prunella vulgaris L. also known as self-heal has been extensively used for treating various ailments in traditional
system of medicine which indicates that the plant is an important source of various bioactive constituents. Current
study was performed to carry out qualitative and quantitative phytochemical evaluation and anti-inflammatory
activity of various extracts obtained from floral spikes of P. vulgaris. Different extracts were obtained from shade
dried floral spikes of P. vulgaris using cold percolation method. The extracts were subjected to suitable
phytochemical tests for the detection and estimation of various phytoconstituents. Aqueous and methanolic
extracts were subjected to inflammatory activity in animal models using carrageenan paw edema method at a dose
of 300 mg/kg of body weight. Diclofenac (10 mg/kg) was used as reference standard. Phytochemical analyses
revealed the presence of flavonoids, tannins, saponins, carbohydrates, steroids, alkaloids and anthraquinone
glycosides. The exact percentage of phytoconstituents obtained by quantitative estimation for alkaloids, saponins,
tannins and carbohydrates was 0.255±0.001, 2.676±0.0141, 0.0988±0.005, 1.584±0.09 per 100 g of plant material,
respectively. Both aqueous and methanolic extracts showed anti-inflammatory activity with maximum percentage
inhibition of inflammation of 28.965 and 30.860, respectively while maximum percentage inhibition of
inflammation shown by Diclofenac was 60.075. Phytochemical analyses revealed the presence various
phytoconstituents which justify its role for the treatment of various ailments in folklore system of medicine.
Besides it can be concluded that both aqueous and methanolic extracts show significant (P<0.05) antiinflammatory
activity comparable to reference standard diclofenac (P<0.01) when compared with control group.
Keywords: Prunell vulgaris, Phytoconstituents, Anti inflammatory, extracts.
... The extracts were prepared by soaking the plant powders in petroleum-ether for 4 days, then the petroleum-ether extracts were filtered by using filter papers. A rotary evaporator apparatus was used to remove the solvent, and the remaining extracts were stored in dark bottles in the refrigerator until use (Harborne 1984). ...
Background
Mulberry silkworm, Bombyx mori L. is one of the most economically important insect because its production of silk which interfere with many important industries. Diseases of the silkworm, B. mori such as viral, bacterial, fungal and protozoan pathogens seriously affect their cocoon production. Bacterial and fungal pathogens independently cause the highest cocoon loss, so the use of botanicals is considered an important strategy to control diseases of silkworm.
Results
The artificially infected silkworm, B. mori larvae with Bacillus thuriengiensis when treated with concentrations of 2 and 3% of black seed and basil leaves extracts increased significantly larval weight and decreased larval mortality. In addition, 3% basil leaves extract and 1% of black seed increased pupal weight. Cocoon weight and cocoon shell weight increased significantly by 3% basil leaves extract treatment. All tested concentrations of black seed extract increased the silk ratios of the resulted cocoons as compared to the infected control. For technological parameters, all tested extracts at 3% concentration caused the highest silk filament length; meanwhile control larvae recorded the highest silk filament weight and size. Regarding the mulberry silkworm, B. mori artificially infested with conidial solution of Beauveria bassiana when treated with all concentrations of the tested plants showed an enhancement in larval weight, pupal weight, cocoon weight, cocoon shell weight, silk filament length, and filament weight and reduced the larval mortality percentage. The highest silk filament size were recorded for the control group.
Conclusions
Tested concentrations of Morus alba, Ocimum basilicum leaves extracts and Nigella sativa seeds extract have suppressed the bacterial and fungal disease spread when used three times through both 4th and 5th larval instars. Moreover, the biological characters and technological traits enhanced by using of these extracts, so it can be used in sericulture for improving cocoon crop quality and quantity.
... The shade dried plant materials were mechanically ground to coarse powder and passed through a Willy Mill to get 60-Mesh size and used for physicochemical, phytochemical and fluorescence analysis. Samples were stored in the good grade plastic containers which are maintained at room temperature until analysis 15 . ...
The macroscopic, microscopic, histological identification and microscopic constants of Salmalia insignis can be used as a rapid, inexpensive and botanical identification technique which would be of immense value in standardization and authentication of this plant. The present study was carried out to investigate the preliminary phytochemical screening and in vitro antioxidant activity of Salmalia insignis. In the present work we investigated the phytochemical screening to find out new sources of natural antioxidant activity source from the leaf of Salmalia insignis with different solvents such as ethanol and petroleum ether. The phytochemical screening was carried out using standard phytochemical tests. Quantitative analysis was performed to confirm and quantify the presence of phenolics content and total flavonoids in the aerial plant extracts of the study plant. The in vitro antioxidant activity was tested on Hydroxyl radical scavenging activity of different solvents such as Petroleum ether, Ethanol and Butylated hydroxynisole. These compounds correspond to varied medicinal properties that can be exploited for the treatment of many diseases.
... The organoleptic evaluation of leaf powder and the extracts, such as colour, texture, odour and taste were carried out as per [8]. The parameters such as actions of plant powder with different chemical reagents, fluorescence analysis, total ash, acid insoluble ash, water soluble ash and extractive values of petroleum ether, RJPBCS 9(2) Page No. 381 ethanol and water were studied according to the official method [8,11]. The determination of moisture content (Loss on drying) was followed by [12]. ...
Plants are being highly explored as a major source of medicinal compounds due to the presence of various phytochemical compounds. In the current investigation, a locally used medicinal plant species Acacia cassia was selected and used for analysis of major bioactive constituents of medicinally important plant. The plant A. cassia belongs to the family Mimosaceae is a glabrous, woody straggling shrub growing up to 6-10 m height. The local name of the species is 'Kari induct or Inhaul'. The plant species is used in Ayurveda, Sidda and Folk medicine. It is commonly known as 'Cabool' in India and ethno medicinally have long been used for the treatment of skin, sexual problems, wound, and stomach and tooth problems. The current study deals with the macroscopic, physic-chemical along with phytochemical analysis and fluorescence analysis of powdered crude drug were carried out for systemic identification and authentication of aerial plant parts. The different extracts of Acacia cassia was studied for its in vitro antioxidant activity using hydroxyl and superoxide anion radical. This study provides referential information for identification and characterization of A. cassia and its extracts.
... For each extract, a phytochemical screening was performed testing the presence of secondary metabolites: tannins, flavonoids, alkaloids, saponins, coumarins, quinones, anthraquinones and terpenoids. The tests were based on the visual observation of colour change or formation of a precipitate after the addition of specific reagents according to Trease and Evans (2002) and Harborne (1998). The qualitative results are expressed as (+) for the presence and (−) for the absence of phytochemicals. ...
The aim of this study was to investigate for the first time the antioxidant and the antimicrobial activities of organic extracts from Traganum nudatum Delile using in vitro models. The extracts which exhibited the highest activities were then submitted to RP-HPLC-PDA analysis. The phytochemical screening revealed the presence of flavonoids, tannins, alkaloids, saponins and quinones. The highest amounts of polyphenols, flavonoids and flavonols were recorded in ethyl acetate fractions. The strongest DPPH scavenging ability was attributed to ethyl acetate fraction of crude extract prepared by maceration EAFM with IC 50 value of 20 ± 0.11 µg/mL, ethyl acetate fraction of crude extract prepared by infusion EAFI was found to possess the highest ferric reducing power with EC 50 of 35 ± 0.4 µg/mL. The highest antioxidant capacity 200.21 ± 0.2 mg AAE/g was exhibited by n-butanol fraction of crude extract prepared by maceration BFM. The antimicrobial activity was proven against all the tested microorganisms except on Candida albicans strains. The extracts were more active on Gram positive than Gram negative bacteria. The strongest inhibition was provided against Bacillus subtilis, Bacillus cereus and Listeria monocytogenes. The identification of phenolic compounds using RP-HPLC-PDA technique revealed the presence of sinapic, syringic, ferulic, p-coumaric acids, catechin, naringenin and vanillin that may be responsible for the bioactivities of the reported specie. Our results indicate that T. nudatum Delile may provide a rich and novel source of natural antioxidants for industrial utilization replacing synthetic ones.
... The phytate content was determined by the spectrophotometric method of Pearson (1976) and expressed in mg/kg. The tannin and phenol contents were determined as described by Harborne (1973) while the dhurrin content of the malts was determined by the method of Eustace and Dorothy (2000). ...
This investigation determined how changes in chemical composition of malting red and white kaffir sorghum grains dried under the sun affected the resistance of the malts to fracture as determined by the Monsanto Hardness Tester. The sorghum grains were malted by a modification of the 2-step wet steep method and dried under the sun. The results showed that malting increased diastatic activity to peak values of 78.40°L and 56°L in the red and white malts; moisture content from 115 to 165 g/kg and 125 to 170 g/kg, crude protein from 104.20 to 183 g/kg and 92.8 to 153.20 g/kg respectively but decreased the fat content. Grain hardness decreased from 83.20 to 42.50 N in the red malts and from 72.70 to 39.30 N in the white. The increases in diastatic activity, moisture, crude protein and reduced fat contents appear to have contributed most to the reduction in grain hardness.
... Chromatographic studies were carried out. 33,34 HPTLC studies 35 were carried out using Camag HPTLC system equipped with Linomat V applicator, TLC scanner 3, Reprostar 3 with 12bit CCD camera for photo documentation, controlled by Win CATS-4 software were used. All the solvents used were of HPTLC grade obtained from MERCK. ...
Aim: The aim of this study was to investigate the aldose reductase inhibition and anti-cataract property of W. volubilis root extract in rats with streptozotocin induced diabetes mellitus. Methods: Diabetes was induced by a single intraperitoneal injection of streptozotocin at the dose of 45 mg/kg. Diabetic animals were treated with glimepiride (0.5 mg/kg) and alcohol extract at 100 and 200 mg/kg for 38 d. Lens were isolated and certain markers were estimated. Results: Treatment of diabetic animals with alcohol extract of W. volubilis root inhibited aldose reductase activity (p < 0.001) and sorbitol formation in the lens. Additionally, a reduction in the levels of MDA, GSH, SDH and LDH in extract treated groups was observed. There was an overall decrease in the severity of cataract progression. Conclusion: The result provided evidence that alcohol extract of W. volubilis root prevents the development of diabetic cataract in rats. © 2019, Association of Pharmaceutical Teachers of India. All rights reserved.
... The aqueous ethanolic extract and fractions of A. indicum were tested for detection of carbohydrates, proteins, alkaloids, glycosides, saponins, phenols, flavonoids and steroids [10,11] . ...
... The phytochemical tests were performed by the method given by Harborne, 1973. ...
Eucalyptus has been used as a medicinal plant from ages because of its various properties. The phytochemical content of Eucalyptus leaves was analyzed by soxhlet extraction of the dried leaves using methanol and acetone. Maximum amount of phytochemicals present in eucalyptus leaves were retained in methanolic extract. The methanolic extract contained quinones, saponins, carbohydrates, tannins, phenols, flavonoids & fat while in the acetonic extract quinones, flavonoids & fat was present. The quantitative analysis of methanolic extract revealed an appreciable amount of antioxidant activity assessed by DPPH scavenging activity expressed in terms of ascorbic acid i.e. IC50 value of 444.6, the phenolic content was found to be 11.41% while the total flavonoid content was 35.88%.
With increasing carbon emissions, environmental problems such as global
warming, melting glaciers, and rising sea levels have increased prominently.
Nowadays reducing carbon emissions and achieving carbon neutrality has become a common goal for all countries. Blue carbon ecosystems (BCEs), including mangrove forests, seagrass meadows and tidal marshes, have more efficient carbon storage compared to terrestrial forests and provide co-benefits like coastal protection and fisheries enhancement. Blue carbon sequestration has therefore been suggested as a natural climate solution. Blue carbon ecosystems cover only 0.2% of the ocean while composing 50% of the carbon burial of the marine sediments. Among all, mangroves can be found in over one hundred countries and territories in the tropical and subtropical regions of the world. Under the combined action of its growth and microorganisms, mangroves capture, transform and store CO2 in the atmosphere into coastal sediment for a long time, and export some organic carbon from the coastal zone to the offshore and ocean. Mangroves provide natural coastal defenses against climate change impacts such as sea-level rise, storm surges, and extreme weather events. Their dense root system helps to stabilize shorelines, reduce erosion and protect inland areas from flooding. By recognizing the invaluable role of mangroves in blue carbon sequestration, policymakers, conservationists, and stakeholders can develop effective strategies for the conservation and restoration of these critical coastal ecosystems, thereby enhancing climate resilience and fostering sustainable development in coastal regions worldwide.
Introduction: Alysicarpus vaginalis belongs to the Fabaceae family, commonly known as buffalo clover and one-leaf clover. Alysicarpus vaginalis is medicinally used in disease-related kidney, diuretics, leprosy, pulmonary problems, jaundice, skin problems, respiratory difficulties, and as a hepatoprotective. Alysicarpus vaginalis is a medicinally important plant according to our review of literature on pharmacological activities, modern pharmacological activities, ethnomedicinal surveys, and traditional systems. However, very limited research related to phytochemical investigation by chromatographic techniques was studied. Methods: So, to investigate the phytochemicals of Alysicarpus vaginalis methanol extract by High-performance thin-layer chromatography and gas chromatography-mass spectrometry. Result: Phytochemical screening revealed the presence of steroids, flavonoids, polyphenols, terpenoids, carbohydrates, alkaloids, and tannins. GC-MS revealed, the presence of six out of nine phytochemicals found medicinally important. 4-O-Methylmannose, Undecane, Neophytadiene, 2-amino-5-[(2-carboxy) vinyl]- Imidazole, Lup-20(29)-en-3-one and stigmasterol are some of the important phytochemical useful as anticancer or as anti-inflammatory. A solvent system was developed by thin-layer chromatography, followed by the HPTLC analysis using the mobile phase toluene: methanol: ethyl acetate (7:3:1). High-performance thin-layer chromatography fingerprinting detected total of 8 peaks with Rf value of 0.007, 0.089, 0.142, 0.211, 0.468, 0.585, 0.772, and 0885 at 254nm and total of 9 peaks with Rf value of 0.007, 0.089, 0.140, 0.208, 0.468, 0.661, 0.772, 0887, and 0.940 at 366nm, respectively. Conclusion: Chemoprofiling by sophisticated spectral techniques such as GC-MS and HPTLC fingerprinting can serve as an important tool for quality control of assuring purity, safety, quality, and efficacy of herbal formulation, and herbal extracts from Alysicarpus vaginalis.
The marine ecosystem harbors unique and diverse bioactive compounds that can offer a vast repertoire of molecules with therapeutic properties. In the present study, four different species of red marine seaweeds were analyzed for its phytoconstituents and the potent antioxidant and anti-inflammatory activity of the methanolic extracts were screened and determined. The results revealed that, among the 4 samples, G. corticata, scored a good antioxidant potential by DPPH (67.61 ± 1.23%, IC50 = 577.7 µg) and metal chelation assay (29.40 ± 0.32%, IC50 = 1684 µg). The anti-inflammatory analysis has shown that, H. dialata was found to exhibit maximum inhibition against the albumin denaturation (83.50 ± 0.24%), whereas G. corticata was observed to measure a maximum inhibition in heat-induced hemolysis (60.40 ± 0.46%) and proteinase inhibition assay (83.30 ± 0.18%). An extensive literature survey was carried out for the bioactive compounds in G.corticata; it was examined for drug likeliness by ADME analysis and toxicological parameters. Further, the best selected bioactive compounds were subjected to in silico molecular docking with pro-inflammatory target, cyclooxygenase (COX-2). Hexadecanal and Neophytadiene were reported to obtain the highest binding affinity (−5.3) for COX-2 enzyme. Hence, in silico molecular docking studies had shown that G. corticata was found to possess potential anti-inflammatory activity that can prevent conversion of arachidonic acid to prostaglandins by inhibiting COX-2. In addition, molecular dynamic simulation studies have shown the stability of Hexadecanal-6 COX complex. To conclude, the outcomes of the present study may shed light on the understanding of the usage of bioactive compounds for therapeutic purpose.
Phytochemical analysis and antioxidant activity in leaves of Cassiafistula, Cassia javanica, Cassia siamea and Cassia suratensis were performed. Dried leaves of C. species were extracted in different polarity gradient solvents for qualitatively of primary and secondary metabolites. Quantitative estimation of phenolics and flavonoids was carried out using FCR and antioxidant activity by DPPH scavenging assay. Phytochemical screening reveals that maximum metabolites are present in ethanol extract in all species as compared to other solvent extracts. Maximum phenolic content was detected in leaves of C. suratensis (200 ± 0.046 GAE/g) followed by C. siamea, C. fistula and C. javanica, respectively. Highest flavonoids were present in C. suratensis and C. siamea (75 ± 0.025 QE/g), respectively. Percentage inhibition (0.02–0.10 mg/ml) was found in C. suratensis (39.72 ± 0.020–91.31 ± 0.001 mg/ml) that is significant. Cassia suratensis was found to be a good source of antioxidants for preventing oxidative stresses-related disease and utilization in therapeutic uses.
Diabetes mellitus is a major health issue that has posed a significant challenge over the years. Gymnanthemum amygdalinum is a well-known plant that can be potentially used to treat this disease. Therefore, this study aimed to evaluate the inhibitory effect of its root, stem bark, leaves, and flower extracts on alpha-glucosidase using an in vitro inhibition assay to isolate the bioactive compounds and determine their levels in the samples. The air-dried plant parts were extracted by maceration using methanol. The results showed that the flower extract had the greatest inhibitory effect (IC50 47.29 ± 1.12 µg/mL), followed by the leaves, roots, and stem bark. The methanolic flower extract was further fractionated with different solvents, and the ethyl acetate fraction showed the strongest activity (IC50 19.24 ± 0.12 µg/mL). Meanwhile, acarbose was used as a positive control (IC50 73.36 ± 3.05 µg/mL). Characterization based on UV, 1H-, and 13C-NMR established that the ethyl acetate fraction yielded two flavonoid compounds, namely, luteolin and 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-methoxy-4H-chromen-4-on, which had IC50 values of 6.53 ± 0.16 µg/mL and 39.95 ± 1.59 µg/mL, respectively. The luteolin levels in the crude drug, methanolic extract, and ethyl acetate fraction were 3.4 ± 0.2 mg (0.3%), 32.4 ± 0.8 mg (3.2%), and 68.9 ± 3.4 mg (6.9%) per 1 g samples, respectively. These results indicated that the G. amygdalinum flower extract exerted potent inhibitory alpha-glucosidase activity.
Context:
Anti-tubercular drugs (ATDs) mediated adverse drug reactions are major concerns for clinicians to treat tuberculosis infection. This study aimed to investigate Andrographis paniculata extract-based phytotherapy to combat the nephrotoxic effects caused by ATDs therapy.
Methods:
Reno-protective effect of A. paniculata extract in ATDs-induced rats was evaluated through LPO, GSH, CAT, SOD, GST, urea, creatinine, uric acid, and histopathological studies. Standardization of the extract was performed RP-HPLC and FTIR analysis. Whereas, the effect of A. paniculata extract on ATDs induced genetic perturbation was analyzed using micronucleus assay. Moreover, the expression level of the xenometabolic gene was investigated using RT-PCR to explore the therapeutic mechanism.
Results:
The nephrotoxic effect of ATDs was indicated by elevated levels of LPO and renal function markers along with the reduced activity of renal antioxidants. An up-regulated expression profile of NAT gene and histological alterations were observed in renal tissue however, micronucleated PCEs were observed in bone marrow cells. Concomitant treatment with A. paniculata extract revealed a noticeable amelioration of elevated oxidative stress markers, gene expression levels, genotoxic perturbation, and histological alterations in a dose-dependent manner.
Conclusion:
Hence, the present study using A. paniculata leaf extract confirmed to play effective phytotherapy against ATDs induced renal toxicity.
The aim of present study was to determine the antioxidant and anti-mycotoxin activities of crude ethanolic extracts from various organs of Bunium ferulaceum Sm. plant from Western Algeria. Firstly, we proceed to a phytochemical screening of leaves, kernels, and barks of plants. Tests showed the presence of several families of chemical constituents and phenolic compounds. The quantitative estimation of the total phenol content, flavonoids, and coumarins is carried out by colorimetric methods. The important total phenolic contents were found in leaves extract with a TPC value of 51.22 ± 0.46 mg GAE/g, whose flavonoids were revealed to be the main phenolic compounds in this extract and the coumarins in bark extract. The antioxidant activity was estimated by the DPPH method compared to the ascorbic acid antioxidant. The leaves extract showed a good scavenging activity with an IC 50 value of 0.11 ± 0.57 mg/ml. Anti-mycotoxin activity are tested against three pathogens fungi produced mycotoxins using mycelial biomass assay and mycotoxigenic test. For all fungi tested, leaves extract showed the most inhibitory activity against Penicillium expansum MTTC 1344 and Aspergillus flavus MTTC 2799 followed by bark extract.
The antibacterial activity of different crude extracts of different parts of Theraxicum officinale including roots, stem and leaves and solvents (n-hexane, chloroform, ethyl acetate and methanol) against various human pathogens viz., Salmonella thphi (S. thphi), Escherichia coli (E. coli), Pseudomonas aeuroginosa (P. aeuroginosa). Staphylococcus aureus, were carried out. Among the extracts, methanolic extract of the roots fraction showed a maximum zone of inhibition (18 mm) against S. aureus, followed by leaves extract of maximum zone of inhibition (16 m), while the ethyl acetate fraction of leaves showed (15 mm) maximum zone of inhibition against S. auresus. While low activity of the stem extracts were recorded except for the methanolic extract with a maximum of zone of inhibition (13 mm) against S. typhi. In addition, the phytochemical analyses showed the presence of Phenols, Tannin, Alkaloids, Saponins and Flavonoids.
The floral source, physicochemical, microbiological, anti-nutritional and phytochemical characterization of four different honey samples obtained from different locations in Ondo State were determined. This was with the view of authenticating their nutritional makeup as was as determine their potential to pose health hazards amongst the consumers. Physicochemical analysis gave moisture (13.5± 0.02-15.0±0.03%), Ash (0.5±0.02%-1.5±0.01%), pH (4.33±0.03-5.28±0.02) Titrable acidity (0.19±0.01-0.47±0.02%), total sugar (0.21±0.01-0.03±0.03%), dextrose (0.21±0.01-0.29±0.02%), fructose (0.22±0.03-0.31±0.01%), hydrated lactose(0.28±0.1-0.04±0.02%), hydrated maltose (0.34±0.03-0.48±0.01%), anhydrous lactose (0.27±0.01-0.48±0.01%), electrical conductivity (0.39x 0²± 0.02-5.21x 10²±0.02ms/cm). Total phenol content from (1.45 to 1.66mg GAE/100g), Total flavonoid content from (0.04-0.07mg QE/100g) and not detected in samples EF3 and GH4. Alkaloid content ranged from (0.06-0.24%), Phytate content ranged from (2.06-3.91mg/100g), Tannin content ranged from (1.37-1.76 mg/100g), Oxalate content ranged from (0.18-0.27mg/100g), Saponin content ranged from (0.06-0.225) and the Ferric Reducing Antioxidant Power ranged from (0.04-0.05mg/100g) and not detected in sample CD2 and GH4. Also, the microbial analysis results revealed that the honey samples had bacterial count between 2.0 x 103- 9.0 x 103 cfu/ml, the total yeast/mould count had a count of 1.0 x 103, 3.0 x 103, 10.0 x 103 and 5.0 x 103 cfu/ml for the four samples respectively. The coliform count indicates the absence of coliform bacteria in all the samples. Isolates from the honey samples were seven different genera of bacteria (Streptococcus, Enterococcus, Pseudomonas, Bacillus, Klebsiella, Micrococcus and a trace of Clostridium spp) which could be hazardous to health if not properly handled.
Powders and methanolic extracts of Clerodendrum capitatum, Phyllanthus fraternus and Tithonia diversifolia were evaluated for their insecticidal activities against Callosobruchus maculatus (Fab.). Plant materials were tested for contact toxicity, oviposition and adult emergence at rates of 0.5, 1.0, 2.0, 4.0 and 5.0 g (w/w) for plant powders and 0.1, 0.2, 0.5, 1.0 and 2.0 ml/20 g (v/w) of cowpea seeds for plant methanolic extracts. The results of the plants powders showed that T. diversifolia was the most effective as it caused 55.0, 60.0, 67.5, 100.0% and 100.0% mortality at the concentrations of 0.5, 1.0, 2.0, 4.0 and 5.0 g/20 g of cowpea seeds within 24 h of application. The least effective plant powder was C. capitatum which evoked the insect mortality of 77.50% at rate 5.0 g within 24 h of exposure. Similar trend of results were obtained on the plant methanolic extracts. Extracts caused more mortality, prevented oviposition and emergence of adult cowpea beetle, more than the plant powders. The calculated lethal dose (LD50 and 90) and concentrations (LC50 and 90) of the plant powders and methanolic extracts showed that T. diversifolia had the lowest values while C. capitatum had the highest across all period of exposure. Tithonia diversifolia methanolic extract completely inhibited oviposition and adult emergence at concentrations 0.2–2.0 ml. Tithonia diversifolia powder and methanolic extract were found to be the most effective in protecting cowpea seeds against C. maculatus. This can be a better alternative to synthetic insecticides since it is abundant in our environment.
Objectives
Alcohol consumption causes several harmful effects on the organs, which is hugely understated. Many deformities occur in the fetus when pregnant mothers indulge in alcoholism. Alcohol is a known teratogen, hence organ formation, particularly development of parts brain critical for cognitive function may be affected. The oxidative brain damage also could contribute to reduced cognitive efficiency of brain exposed to alcohol. In this study, effect of Centella asiatica in relieving the oxidative brain damage in offspring of alcohol fed mother rats was evaluated.
Methods
In this study we fed alcohol (5 g/kg body weight, 30% w/v) to a group of pregnant Wistar rats during gestation period, and another group served as control. Four groups of rats (n = 6 each) were selected from the offspring of these mother rats. The groups were, control, positive (treated) control, untreated and treated from alcohol-fed mother. Their cognitive parameters were tested in water maze, shuttle box and compared. Further their oxidative status was evaluated by estimating malondialdehyde (MDA), protein carbonyl, total antioxidants and glutathione reductase (GSH) in hippocampus.
Results
The results suggested that there was significantly high cognitive performance in maze test and shuttle box memory retention in rats treated with C. asiatica water extract and the antioxidant levels were high in their hippocampus.
Conclusions
The outcome of the study suggested that C. asiatica produced beneficial effects in reversing the alcohol induced brain damage in pregnancy.
Plant metabolomics deals with the interpretation of various metabolic pathways in contrast to other -omics technologies applied in systems biology. Metabolomics is a highly challenging field where the metabolite analysis is done by high-end technologies for proposing metabolic pathways. The high-throughput technologies utilized for these studies importantly include mass and nuclear magnetic resonance spectrometry. Both these techniques have their own specific features and it is usually difficult to interpret the data compared to genomics and transcriptomics data. The metabolic pathways in plants are changed during biotic and abiotic stresses. These changes can be noted at each step through metabolite analysis. Plants acclimatize to the changes during stress conditions by producing secondary metabolites by other mechanisms. We may analyse these changes by advanced techniques.
Seven-day-old maize seedlings were subjected to NaCl (150 mM) without or with the presence of equimolar concentrations of urea, potassium nitrate or ammonium sulfate (at 60 kg N ha−1) for the subsequent 15 days. The results show that NaCl significantly decreased photosynthetic activity parameters (Hill reaction, Pn, Ci and Gs), shoot fresh and dry weights, chlorophylls, protein, GSH and AsA. The activities of ALA-D, Rubisco, PEP-C, MDH, PPDK, SOD, CAT and APX were significantly inhibited. On the contrary, NaCl significantly elevated MDA, O
∙−2
, H2O2, soluble sugars and proline. The application of the nitrogenous sources mostly ameliorated the photosynthetic activity synchronous with counterbalancing the decreases in growth, protein and pigments. The activities of ALA-D, Rubisco, PEP-C, MDH and PPDK enhanced remarkably with N applied under salinity. Furthermore, enhancements were detected in GSH and AsA contents and SOD, CAT and APX activities, which decreased free radicals, soluble sugars and proline. These effects reveal that maize was vulnerable to NaCl stress; however, N overcame the stress status and ameliorated the plant tolerance to salinity concomitant with repairing photosynthetic activity and maintaining higher antioxidants and ROS homeostasis; urea seemed to be the most efficient. Taken together, these findings conclude that N enhanced the capability of maize to tolerate NaCl via protecting photosynthetic apparatus for normal photochemical functioning and improving antioxidants and ROS homeostasis as effective salt tolerance mechanisms.
Garcinia Kola is commonly used in traditional medicine for the treatment of diverse ailments including coronary artery diseases. Thus, this study aims to determine the effect of hydromethanol (1:4) extracts of the pulp and seed coat of Garcinia kola on serum lipid profile and its antioxidant properties. The two forms were separately dried and blended to powder. Forty male wistar rats (8 per group) were assigned into Five (5) groups. Groups were treated thus: Group one; control. Group two; 100mg/kg pulp extract. Group three; 200mg/kg pulp extract. Group four; 100mg/kg seed coat extract. Group five; 200mg/kg seed coat extract; for 30 and 60 days duration. On treatment conclusion, blood was collected for the determination of lipid profile and antioxidant properties. The higher dose of the pulp and seed coat extracts significantly (P˂0.05) increased the catalase level and superoxide dismutase enzyme activity, whereas, both the higher and lower doses of the seed coat extract caused a reduction in malondialdehyde level. The serum total cholesterol was significantly elevated by the higher dose of the pulp extract while the seed coat extract caused significantly increased high density lipoprotein cholesterol level and a reduction in the low density lipoprotein level. The two extracts demonstrated marked antioxidant effects. The seed coat of Garcinia kola may possess the potential to prevent cell death due to lipid peroxidation by inhibiting the lipid peroxidation process. The seed coat extract may also be useful in preventing coronary artery disease and other atherosclerotic problems.
The present study was included investigation efficiency of alcoholic and aqueous extract of Artemisia herba-alba in treatment of albino mice infected experimentally with T. muris comparative with metronidazole drug. The doses of alcoholic and aqueous extracts of A. herba-alba for treatment were 750 mg/ kg as a single treatedable dose and then done comparable between efficiency of extracts under study and Metronidazole drug that given as a single dose of 0.392 mg/ mouse/day.
The results of this study revealed efficiency of both metronidazole drug and extracts of A. herba-alba in treatment the infected with T. muris. So these were treated with materials revealed effective and efficiency in treatment (100%), but it were different in the time for reduction of infection and perfect cure. Also the effect investigated of alcoholic and aqueous extract of Artemisia herba-alba and metronidazole drug in intestinal (ileum and colon) and liver, where observed normal form of intestinal and liver tissues of infected mice that treated with alcoholic and aqueous extract of Artemisia herba-alba and metronidazole drug, while observed abnormal form of intestinal and liver tissues of infected mice (positive control) and not treated with alcoholic and aqueous extract of Artemisia herba-alba and metronidazole drug.
The study was conducted to investigate the effect use of water extract and chloroform of peppermint plant Mentha spicata in control of mosquito Culex molestus during the treatment of the fourth instar larvae. Use in the experiment aqueous cold, boiling and chloroform of roots, stems and leaves extracts of Mentha spicata was studies by using concentrations (1,5 and 10) mg/ml of aqueous extracts and (0.25,0.5 and 1) mg/ml of chloroform extract on some biological aspects of Culex molestus by treated fourth instar larvae of the insect, and the results were as follows : The results of the study showed that the above mentioned extracts have an effect in inhibition of emergence and that effect is positively correlated with the concentrations used, the most of these effects were demonstrated by using the chloroform extract of the leaves, as for the effect of the mortality of the fourth instar larvae and pupae, the a cumulative mortality reached 93.9% at the concentration of 0.25 mg / ml and did not differ significantly when the concentration increased to 1 mg / ml, which amounted to 90% we note that the highest percentage of mortality occurred when using aqueous cold, boiled and chloroform for leaves also noted many morphological deformations during the experiments. These extracts may act as insect growth regulators, leading to these deformations.
Soybean seeds, soaked in branched-chain amino acids [Valine, Isoleucine and Leucine (VIL)] mixture or dressed in naphthalic anhydride (NA), were germinated for 10 days and treated with chlorimuron-ethyl for 120 h. Chlorimuron-ethyl significantly inhibited acetolactate synthase (ALS) activity in shoots of all samples during the first 72 h. Moreover, it increased Km but decreased Vmax, Kcat, Kcat/Km and Vmax/Km as well as the endogenous levels of valine, isoleucine and leucine particularly during the first 72 h. In concomitance, there were significant decreases in protein content and activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT) as well as growth parameters during the first 72 h. However, VIL or NA mitigated the herbicide effect. So, the drop in Vmax, Kcat and Kcat/Km by chlorimuron-ethyl declares decreases in enzyme concentration, catalytic rate and catalytic efficiency, respectively. Whereas the increase in Km could indicate that chlorimuron-ethyl induced an inhibition for ALS of the mixed-type which would affect synthesis and structural integrity of the enzyme. This inhibition would diminish the formation of branched-chain amino acids and protein, particularly with the inhibition of N incorporation enzyme (NR, NiR, GS and GOGAT). Consequently, retardation and disturbances in formation of structural and functional protein would arise. Nonetheless, VIL mixture overcame chlorimuron-ethyl toxicity via compensating the drop in the endogenous branched-chain amino acids while NA might cause mitigation by detoxifying the herbicide.
Marine brown algae Turbinaria ornata (T. ornata) that grows along Tamilnadu coast (Gulf of Mannar) were collected to study the composition of major and minor nutrients, phytochemical constituents and to evaluate the cytotoxic effect. Proximate composition of this brown seaweed was investigated by the determination of moisture, ash, protein, lipid and carbohydrate. The mineral content was analyzed using Inductive Coupled Plasma- Mass Spectrometry (ICP-MS). In general, the results of proximate analysis showed that moisture content (83.62%) was most abundant followed by carbohydrate (43.7% dry weight), ash content (23.4%) protein (5.4% dry weight) and lipid (1.9% dry weight). This work presents the data on the mineral content such as manganese, sodium, magnesium, potassium, zinc etc. of the brown algae. The chemical components present in the methanolic extract of the sample were analyzed using Gas Chromatography–Mass Spectrometer (GC-MS). The presence of secondary metabolites illustrate that this brown seaweed T. ornata can be harnessed for their biomedical potentials. Apart from this, the methanolic extract extracted also showed cytotoxic activity against human retinoblastoma Y79 cell lines and the Inhibitory Concentration (IC50) was found to be at 6.37 μg/mL. Further research should be explored for the beneficial application of T. ornata as a potent therapeutic tool against cancer.
Cucumis sativus is a vegetable crop belonging to the Cucurbitaceae family and originates from temperate, subtropical sub-humid areas. This study was aimed to evaluate the effect of water extract of immature melon seeds in inhibition of seed germination of cucumber and celery plant. This study was conducted in the laboratory of the Department of Science, Faculty of Basic Education, Diyala University, Iraq. The seeds of these vegetables were selected and diagnosed in the herbaceous plant in the Department of Life Sciences. During the washing of immature seeds that were isolated from other melon residues. The results showed that the concentration of (0.1, 0.3, 0.7) and compared to the comparison models had the same effect in germination in seeds for cucumber. For celery plant only (0.7) gave the highest percentage of inhibition of celery seed plant and reached 20% Which was 80%. As for the length of the root only, the concentration of (0.7) gave the lowest rate for the cucumber and celery which amounted to 2.132 and 0.866 respectively, compared to the comparison treatment amounted to 5.932 and 1.399 respectively, in addition to other transactions. In addition, the concentration (0.7) gave the lowest length of the cucumber and celery time of 8.799 and 2.566 respectively, compared with the comparison treatment given to both plants 13.232 and 3.066 respectively, in addition to other treatments. As for the soft and dry weight of the cucumber plant, the concentration was given (0.7) the highest rate of soft weight at 0.2920, while the concentration (0.1, 0.3) gave 0.2057 and 0.0599 respectively, while dry weight gave the concentration (0.7).
Vetiver is used for removing heavy toxic metals in groundwater. It is also largely used for medicinal and perfume industry. The main element that we come to know about the Vetiver system is Vetiver grass (Vetiveria zizanioides) is a sterile, non-invasive tropical bunchgrass. We can also use Vetiver (Khus) as a flavoring agent, mainly as Khus syrup, which is produced by adding khus essence to water, sugar and citric acid syrup. Usually, Khus essence is looked as wide dark green in color and it is taken from the roots. It gives a scent and a woody taste distinguishing trait of khus. Another use of Vetiver syrup is to compose Yogurt drinks similar to lassi and Milkshakes, Ice creams, and at last mixed beverages like dessert topping and as a Shirley Temples. It is completely used in a vital level of applications containing with food and pharmaceutical industries. The central purpose of the present research is to discover the phytochemical, antioxidant properties situated in the crude descent of (Vetiveria zizanioides). The methanolic extract as an antioxidant activity reveals a preferable radical scavenging activity than some other selected solvents. FT-IR and GC-MS results point out the presence of Aliphatic, Alkanes includes Aromatic elements chief constituents seemed to be Vetiveria zizanioides. The Conclusion of this study virtually shows that (Vetiveria zizanioides) crude extract could be reflected as health nutrients in pharmaceutical and food industries.
A technique has been developed for the separation of proteins by two-dimensional polyacrylamide gel electrophoresis. Due to its resolution and sensitivity, this technique is a powerful tool for the analysis and detection of proteins from complex biological sources. Proteins are separated according to isoelectric point by isoelectric focusing in the first dimension, and according to molecular weight by sodium dodecyl sulfate electrophoresis in the second dimension. Since these two parameters are unrelated, it is possible to obtain an almost uniform distribution of protein spots across a two-diminsional gel. This technique has resolved 1100 different components from Escherichia coli and should be capable of resolving a maximum of 5000 proteins. A protein containing as little as one disintegration per min of either 14C or 35S can be detected by autoradiography. A protein which constitutes 10 minus 4 to 10 minus 5% of the total protein can be detected and quantified by autoradiography. The reproducibility of the separation is sufficient to permit each spot on one separation to be matched with a spot on a different separation. This technique provides a method for estimation (at the described sensitivities) of the number of proteins made by any biological system. This system can resolve proteins differing in a single charge and consequently can be used in the analysis of in vivo modifications resulting in a change in charge. Proteins whose charge is changed by missense mutations can be identified. A detailed description of the methods as well as the characteristics of this system are presented.
Surmmary
Soluble cell constituents of strains of Streptococcus faecalis, S. faecium and S. durans were studied by electrophoresis in polyaery lamide gels followed by staining the gel to reveal the patterns of separated proteins and esterase enzymes. The patterns of proteins and of esterases found in S. faecalis differed from those in S. faecium and S. durans. Strains of S. faecalis differing in serotype and variety showed a very similar series of proteins, some differences in patterns of esterases were found, but similar patterns occurred in strains differing in type and variety. The major protein bands of S. faecium and S. durans differed in mobility from the major proteins of S. faecalis; different strains of S. faecium and S. durans showed some differences in other protein bands. Esterase activity of S. faecium and S. durans was weaker than that of S. faecalis, and in the case of most strains only faint esterase bands were detected. A strain of S. faecium which gave protein and esterase patterns different from those of the remaining strains, differed also in showing motility.
1. Gels were prepared with recrystallized acrylamide and bisacrylamide. Electrophoresis was in tris-sodium acetate-EDTA buffer for 0.5 to 3hr. Gels were scanned at 280 or 265mmu. Techniques are described for slicing and radioactive counting. 2. The mobility of RNA was inversely related to the sedimentation coefficient and varied with gel concentration. Electrophoresis in 2.2-2.6% gels gives a fractionation similar to density-gradient centrifugation. It shows the two ribosomal RNA components, the 45s precursor, transfer RNA and minor components. In 5% and 7.5% gels, 4s and 5s RNA separated and ribosomal RNA was excluded. 3. The resolution is greater and more detailed than by centrifugation, and many samples can be analysed simultaneously and rapidly.
SUMMARY Hyphal walls of two phytopathogenic moulds, Phytophthora cinnamomi and P. parasitica, were isolated essentially free from cytoplasmic con- tamination. They have a complex chemical structure consisting of poly- saccharide, protein and lipid. D-Glucose was the main monosaccharide detected in acid hydrolysates. Chromatographic evidence suggested the presence of small amounts of mannose (0-6y0), glucosamine (0.3%) and traces of galactosamine and ribose. Glucans constituted nearly 90 yo of the wall but only about a maximum 25 yo of the wall could be regarded as cellulose I on the basis of solubility, resistance to hydrolysis and X-ray diffraction. Most of the wall glucan exhibited chemical and physical properties unlike typical cellulose. The spectrum of amino acids commonly found in fungal walls was detected ; hydrolysates also contained hydroxyproline and two minor unidentified ninhydrin-positive components. Protein comprised 3-5 yo of the wall, A small amount of lipid (1-3 yo), mostly of the bound type, was found, and also traces of phosphorus, and compounds with absorp- tion maxima at 263mp. Hyphal walls of Phytophthora cinnamomi and P. parasitica differed only slightly in quantitative composition.
The essential features of constitution, configuration, and conformation in carbo hydrate chemistry, so well established in the . first half of this century, had yet to be exploited by those concerned with biochemical and physiological processes in plants when the original Encyclopedia appeared. Two outstanding developments, discovery of sugar nucleotides and the advent of chromatography, brought together the insight and a means of probing complexities inherent in plant carbohydrates. These advances, combined with a modern knowledge of enzymes and cellular metabolism, have provided new horizons of investigation for the student of plant physiology. This volume and its companion (Vol. 13B) present a comprehensive assess ment of the current viewpoint in plant carbohydrates with emphasis on those aspects which impinge on physiological processes of growth and development. To accommodate the extensive amount of information to be presented, subject matter has been divided, somewhat arbitrarily, into intracellular and extracellular carbohydrates, with the latter defined as carbohydrates occurring in space out side the plasma membrane (plasmalemma). This classification is not exclusive; rather it is intended to lend a degree of flexibility to the way in which subject matter is arranged between volumes. The first section of this volume addresses the occurrence, metabolism, and function of monomeric and higher saccharides of fungi, algae, and higher plants.
In 1958, a single volume in the original series of this Encyclopedia adequately summarized the state of knowledge about plant carbohydrates. Expansion into two volumes in the New Series highlights the explosive increase in information and the heightened interest that attended this class of compounds in the interven ing years. Even now the search has just begun. Much remains to be accom plished; e.g., a full description of the plant cell wall in chemical terms. Why this growing fascination with plant carbohydrates? Clearly, much credit goes to those who pioneered the complex chemistry of polyhydroxylated compounds and to those who later sorted out the biochemical features of these molecules. But there is a second aspect, the role of carbohydrates in such biological func tions as host-parasite and pollen-pistil interactions, the mating reaction in fungi, symbiosis, and secretion to name a few. Here is ample reason for anyone concerned with the plant sciences to turn aside for a moment and consider how carbohydrates, so many years neglected in favor of the study of proteins and nucleic acids, contribute to the physiological processes of growth and devel opment in plants.
Biochemical studies on leaf proteins carried out by Wildman and Bonner (1947) revealed the presence of a major protein component having a large molecular size (18s) which was designated as fraction-1-protein. The ubiquitous distribution of this protein in green plant leaves and green algae, as determined by analytical ultracentrifugation and immunological precipitation methods, stimulated later studies on its enzymic nature (Dorner et al., 1958). The independent investigation of the path of carbon in photosynthetic CO2 fixation, together with that on the enzymic machinery of the reductive pentose phosphate cycle, led to the discovery of ribulose-1,5-bisphosphate (RuBP) carboxylase (E.C. 4.1.1.39; carboxydismutase) catalyzing the following key reaction [Eq. (1); Quayle et al., 1954; Weissbach et al., 1954, 1956; Chap. II.1, this vol.] (1)
Isoelectric focusing is an electrophoretic method that utilizes the migration behavior of amphoteric molecules in a pH gradient to achieve their condensation into narrow isoelectric zones that are stationary in the electric field. The steady-state position of each zone in the pH gradient depends on the isoelectric point (PI) of a particular amphoteric molecule, and therefore, isoelectric focusing can be used as a separation technique. The ampholyte that has the lowest pI migrates closer to the anode, where it is condense in its isoelectric state at some distance from the anode because of repulsion. The same process condenses the most basic ampholyte close to the cathode. Carrier ampholytes with intermediate pI values is focused at different positions along the electric field. Uniform conductance in the pH gradient is also important for several reasons. Regions of low conductance exhibit higher electric field strengths and therefore produce more Joule heat, which may cause convective disturbances.
The use of chromatographic and electrophoretic techniques for the separation of proteins has an immense impact on the progress of modern biochemistry. Because proteins exhibit diverse biological functions (e.g., enzymatic, hormonal, regulatory, and immunological functions) there is no field in the biological and biomedical sciences where chromatography and electrophoresis have not been utilized advantageously, either for research or clinical purposes. The separation of proteins requires caution in regard to the pH, ionic strength, temperature, and nature of the electrolytes and supporting material to be employed. The physico-chemical and biological properties of each particular protein depend on a combination of all of these variables. The three levels of structure (viz., secondary, tertiary, and quaternary) and molecular interactions depend on ionic, hydrophobic, and hydrogen bonding. The same forces are also involved in the separation processes. Thus, the environmental physical conditions have to be chosen in such a manner that the separated product maintains certain desirable properties, usually associated with the preservation of the native state and biological activity.
This chapter focuses on thin-layer, ion-exchange, affinity chromatography and paper and gel electrophoresis of nucleic acids and their monomeric constituents. It discusses the use of chromatography and electrophoresis in sequence analysis and nucleoprotein separation. The base composition of an unknown RNA or DNA molecule is obtained by first degrading the polymer to purine and pyrimidine bases, nucleosides, or nucleotides, separating the individual species, and estimating the proportion of each. The advantage of nuclease degradation is that products can usually be applied directly to ion-exchange columns or thin layers without the neutralization or removal of the reagent. Chemical degradation tends to destroy many of the modified sugar residues found in tRNA species. The common purine and pyrimidine bases possess ring nitrogen atoms and, with the exception of thymine (DNA) and uracil (RNA), exocyclic amino groups that accept protons. Hence, by changing the pH of the solvent, the degree of protonation on each type of base can be controlled.
Brief red light treatment causes a temporary rise in the rate of cytoplasmic rRNA synthesis in etiolated pea seedling buds and a temporary fall in excised epicotyl sections. In each case a lag of 1–3 hr occurred before the changes were detectable. No changes could be detected in rRNA synthesis in excised apical sections of dark-grown oat coleoptiles. By depressing rRNA synthesis selectively with 5-fluorouracil it was possible to show that red light treatment did not cause any changes in the overall rate of polydisperse RNA synthesis. Neither 5-fluorouracil nor actinomycin D selectively inhibited the red light induced changes in growth in pea and oat sections. The results suggest that phytochrome does not operate through the regulation of transcription.
A possible net movement of nitrogen between peel and pulp tissues of ripening bananas was evaluated. No evidence of a movement of nitrogen from the peel to the pulp was found, nor with either tissue did the proportion of the total nitrogen extracted by 5% (w/v) trichloracetic acid change significantly during the climacteric period. Neither the content of RNA in the pulp tissue, nor the base composition of the RNA changed during the climacteric. It is concluded that no substantial increase in the amount of ribosomal material accompanies the increase in the rate of protein synthesis early in the climacteric.
The carbohydrate compositions of mung bean (Phaseolus aureus) shoots and particulate preparations obtained from these plants, which synthesize polysaccharides in vitro, have been compared. Both types of tissue contained hot water soluble polysaccharides (starch, galactan and arabinan), pectin, hemicellulose and cellulose, but generally at much lower levels in the particulate preparations. Whole shoot hemicellulose contained d-galactose and d-glucose as the major monosaccharides, with a lesser amount of d-xylose and arabinose, and only traces of d-mannose. In contrast, the particulate hemicellulose contained d-glucose, d-galactose, arabinose and d-mannose in significant amounts, but only traces of d-xylose. The major lipid-bound sugar in the tissues was d-glucose, but lipid-bound d-galactose, l-rhamnose, and possibly arabinose and d-xylose were also present. A partial acid hydrolysis of the particulate preparation yielded the following oligosaccharides: cellobiose, 6-O-β-d-galactosyl-d-galactose, 3-O-β-l-arabinosyl-l-arabinose, di- and tri-d-galacturoni acids, d-galacturonosyl-l-rhamnose, d-glucuronosyl-d-galactose and d-glucuronosyl-d-galactosyl-d-mannose.
The albumin and globulin fractions of the seeds of Brassica campestris, B. oleracea, and B. nigra have been investigated using serological methods and separation by acrylamide gel electrophoresis in order to determine the
usefulness of these techniques for taxonomic studies of the genus Brassica. The results of the protein analysis given in this paper agree with established taxonomy and suggest that both methods may
prove useful in various taxonomic problems in the genus Brassica.
The globulins extracted from a number of different species of legume seeds have been subjected to disc electrophoresis. In the limited sample surveyed, the tribes Vicieae, Phaseoleae, Diocleae, Trifolieae and Daibergieae were each characterized by a distinctive protein band pattern. When the globulins of genera from the Loteae, Galacteae and Glycineae were examined, two types of protein band patterns were obtained which were distinctive for each tribe. A wider range of protein band patterns were obtained when genera of the Cajaneae were examined. The one species of Abrus examined possessed a globulin protein band pattern which differed from the patterns found in any other genera of the tribes examined.
If the findings reported here can be extended to a much greater range of genera without significant change, then for some tribes within the Leguminosae a distinctive protein band pattern can be obtained when their seed globulins are electrophorized.
Ribulose-1,5-bisphosphate carboxylase/oxygenase has been purified to electrophoretic homogeneity from comfrey, Symphytum spp. Sodium dodecyl sulfate polyacrylamide and polyacrylamide gel electrophoresis studies on the purified product showed no extraneous proteins. Comparisons of the electrophoretic mobilities of the subunits to those of standard proteins indicated a large subunit MW of 50 000 and a small subunit of 12 700, which for an octameric structure of each subunit indicates a native MW of 502 000. Specific activities of the comfrey enzyme ranged from 1.2 to nearly 2 μmol 14CO2 fixed/min.mg of protein over several preparations and were maintained for months when stored from the sucrose gradient at − 70°. The specific activities depended critically on the amounts of enzyme used in the assay even under saturating conditions of substrates and cofactors. The effective pH dependence for carboxylase catalysis peaked near 7.4, which apparently is the lowest elective optimum yet reported for this enzyme from any source. However, on a constant carbon dioxide basis the pH dependence profile was reversed with a maximum near pH 8.6 which was 0.4 units higher than the value for the spinach enzyme. The Kms for carbon dioxide and ribulose-1,5-bisphosphate at pH 7.5 were 130 μM and 30 μM, respectively, which are comparable to the accepted values for the carboxylase from spinach at pH 7.2.
The composition of corresponding linear and branched polymers from the hemicelluloses of three Gramineae: Lolium perenne (grass), Triticum vulgare (wheat) and Zea mais (maize) and three Leguminosae: Trifolium pratense (red clover), Medicago sativa (lucerne) and Glycine max (soya bean) were compared.The corresponding polymers from the three species of Gramineae were very similar as were those from the Leguminosae. However there was a distinct difference between the two plant families. The linear polymers of the Gramineae contained more arabinose and less glucuronic acid than those from the Leguminosae. The greatest difference between the two plant families was found in the branched polymers, those from the Gramineae containing a high percentage of xylose and rather small amounts of arabinose, galactose and uronic acid, whereas those from the Leguminosae contained relatively large amounts of uronic acid, galactose and arabinose and little xylose. In the branched polymers from the Gramineae the uronic acid was linked to xylose, whereas in those from the Leguminosae it was linked to arabinose.
The methods available for the extraction and estimation of nucleotides and nucleic acids have been applied to a variety of plant tissues. It has been shown that, with the tissues investigated, either aqueous ethanol or dilute perchloric acid could be used to extract the soluble nucleotides. Components other than nucleotides contributed to the 260 μu optical density of the extract, and with most tissues only 20 to 30 per cent of the optical density was due to nucleotide material. Although purification of the nucleotides in the extract was attempted by various procedures, such as precipitation of the nucleotides as insoluble salts, extraction of pigments with organic solvents and ion-exchange treatments, no suitable general method was found. The separation into individual nucleotides by ion-exchange chromatography was the only reliable way of estimating the nucleotide content of a tissue.The Schmidt and Thannhauser procedure was the only general method suitable for the extraction of nucleic acids from plant tissue. Purification of the RNA mononucleotides before estimation was essential, but the anion-exchange treatment described by Smillie and Krotkov did not remove all the material which interfered with the estimation of the RNA by phosphate or ribose content. The separation of the mononucleotides by ion-exchange chromatography provided the most reliable method of estimation. The DNA content of the tissue as estimated by the Schmidt and Thannhauser method was considerably lower than that obtained by the direct, hot acid extraction of Schneider, due in part to an incomplete precipitation of the DNA after alkaline hydrolysis. The Schneider method was suitable for the estimation of RNA under special conditions, such as in tissue homogenates containing no cell debris or nuclei, while the Ogur and Rosen procedure appeared to be more useful as a method of RNA fractionation than for the quantitative measurement of RNA.
The numbers of cells and chloroplasts, and the amounts of DNA, RNA and chlorophyll in emerged spinach leaves of different age, have been measured. In addition the ribosomal—RNAs have been fractionated by polyacrylamide gel electrophoresis. Cell size, the number of chloroplasts and the quantities of chlorophyll, DNA and ribosomal—RNAs per cell all increased with age. Despite a 3-fold increase in cytoplasmic ribosomal—RNA, and a 15-fold increase in chloroplast numbers per cell, chloroplast ribosomal—RNA per chloroplast remained unchanged as the leaves expanded.
The two distinct soybean trypsin inhibitors, namely Kunitz and Bowman-Birk inhibitors, were isolated from various commercial sources by isoelectric focusing in the narrow pH 3–6 range in sucrose gradients. The Kunitz inhibitor was also isolated by the same procedure from an aqueous soybean water extract subjected to preparative gel filtration on Sephadex G-25. The Kunitz inhibitor was focused at pH 4.5, and the Bowman-Birk inhibitor at pH 4.3. The two purified proteins were found to be immuno-chemically different. Disc electrofocusing on polyacrylamide gels in the pH 3–10 range was shown to be a simple and high resolution method for detection of heterogeneous preparations of the two inhibitors.
Two relatively new electrophoretic methods for the separation and characterization of proteins and other ion species, isoelectric focusing in natural pH gradients and isotachophoresis, are reviewed.
The chloroplast gene coding for the large subunit of ribulose 1,5-bisphosphatecarboxylase in Zea mays has been sequenced. It contains neither a ‘classical’ prokaryotic promoter sequence nor its proposed eukaryotic counterpart but does have a typical prokaryotic ribosome binding site close to the site at which translation is initiated. Almost all possible codons occur and are translated by the ‘universal code’ rather than by the variation of it found in mitochondria.
Total hemicelluloses have been isolated from the leaves and stems of field-grown oat plants and from the leaves, roots and coleoptiles of plants grown in the laboratory. In any one part of the plant, with increasing maturity there is an increase in the percentage of xylose and a decrease in the percentage of glucose residues. These changes and accompanying changes in the percentages of galactose and arabinose residues are considered in terms of three pure hemicelluloses known to be present—an arabinoxylan, an acidic galactoarabinoxylan and a non-cellulosic glucan. Pure galactoarabinoxylans were isolated from the leaves of field-grown oat plants at different stages of growth. The results indicate that it is inappropriate to study the effect of growth on hemicellulosic composition solely by considering pure hemicelluloses.
The three variations of the scheme of analysis proposed in Part I are tested on different roughages and compared with Harwood's sulphuric acid hydrolysis method. The results of the summative analysis on grain straw, legume straw, hay, artificially dried grass, fresh grass, mangold and some crude fibres are given.
Mannose has been measured in hydrolysates of cell wall carbohydrates from fronds of 109 species of ferns from 40 genera. Sixty-four species from 24 genera contained high (10–20%) or medium-high (5–10%) levels of mannose in the cell wall carbohydrates of their green leaf (laminar) tissue. Forty-five species in 16 genera however (principally Thelypteris, Asplenium, Blechnum, Doodia and Polystichum) contained little or no mannose (<2%) in the cell wall carbohydrates from laminar tissues. The few species examined from fern orders other than the Filicales also had low levels of cell-wall mannose. Only one genus, Phymatodes, gave species with high and low levels respectively of polymer mannose in the lamina. Cell-wall mannose levels were also measured in midribs from the low mannose ferns and found to be low only in the Asplenium spp. Holocellulose (11·5 g) from one high mannose species, Pyrrosia serpens, was fractionated with alkali into hemicellulose (1·2 g), possible galactoglucomannan (4·5 g) and cellulose (2·7 g).
A method of extraction of RNA from coffee based on phenol treatment is described. Effectsf of various agents and pH of the extracting buffer on the efficiency of extraction were studied. The best extracting solution is 0·2 M Tris-HCl buffer at pH 7·4 with 1% sodium dodecyl sulphate and 0·05% EDTA. RNA (5–6%) is lost in the tissue residue and 4·6% in the interphase layer. No significant deviation of the spectral characteristics of the RNA solutions obtained from three samples of coffee from that for purified yeast RNA is observed. The purine-pyrimidine ratio for the RNA has been found to be in the range of 1·25–1·38.
Two globulin fractions, obtained by extraction using acidic conditions, were characterized by their sedimentation properties, subunit compositions and
Factors affecting the amount and type of isotopically labelled nucleic acid extracted from peanut cotyledons by a two-phase partition system employing phenol were investigated. Nucleic acids isolated at 37° contained some degraded ribosomal RNA, while at 60° ribosomal RNA was severely degraded and DNA was completely eliminated from the aqueous extract. Changing the pH of the extraction medium over a range between 6·0–8·5 had little measurable effect on the type of nucleic acid extracted. However, addition of dupanol (sodium lauryl sulfate) to the extraction medium increased the amount of nucleic acids by 40 per cent, and the specific activity by 50 per cent; little DNA was isolated without dupanol. Differential extraction techniques involving variations in temperature, pH or dupanol gave two different samples of nucleic acid in each case. One contained most of the extractable nucleic acid while the second contained only 4–5 per cent of the total nucleic acid which was composed mostly (as judged by its relatively high specific activity) of newly synthesized ribosomal and messenger RNAs and DNA.
The ribulose-1,5-bisphosphate carboxylase/oxygenase purified from maize (a C4 monocot) to homogeneity has a MW of532 000 and sedimentation coeffici
The effect of DNA fragment size on the extent of hybridisation that occurs between repeated sequence DNAs from oats, barley, wheat and rye has been investigated. The extent of hybridisation is very dependent on fragment size, at least over the range of 200 to 1000 nucleotides. This is because only a fraction of each fragment forms duplex DNA during renaturation. From these results estimates of the proportions of repeated sequences of each of the cereal genomes that are homologous with repeated sequences in the other species have been determined and a phylogenetic tree of cereal evolution constructed on the basis of the repeated sequence DNA homologies. It is proposed that wheat and rye diverged after their common ancestor had diverged from the ancestor of barley. This was preceded by the divergence of the common ancestor of wheat, rye and barley and the ancestor of oats. Once introduced in Gramineae evolution most families of repeated sequences appear to have been maintained in all subsequently diverging species. — The repeated sequences of oats, barley, wheat and rye have been divided into Groups based upon their presence or absence in different species. Repeated sequences of related families are more closely related to one another within a species than between species. It is suggested that this is because repeated sequences have been involved in many rounds of amplification or quantitative change via unequal crossing over during species divergence in cereal evolution.
Reassociation kinetics of sheared denatured DNAs from wheat, barley, rye, and oats at 60 C in 0.18 cm Na+ indicate that approximately 80% of these genomes consist of repeated nucleotide sequences, using hydroxylapatite chromatography to detect reannealed DNA. The remaining DNA appears to consist of sequences present in only one or a few copies per haploid genome. Studies on heterologous duplexes formed in vitro between the repeated sequence DNA fractions from each of the species in turn indicate that many of the families of repeated sequences in these cereals evolved from common ancestral sequences. The extent of heteroduplex formation and duplex thermal stabilities suggest a scheme for the evolution of these species which agrees with taxonomic and genetic evidence. Further analyses of these parameters indicate that many quantitative changes in the chromosomal complement of repeated sequences have occurred during divergence of these species.
Some 300 umbellifer species, representing 52 per cent of the genera of the family, have been surveyed for their leaf phenolics, using both fresh and herbarium tissue. The results show that, with few exceptions, species can be divided into two groups, those with flavone (usually luteolin) and those with flavonol (kaempferol and/or quercetin). These groupings are mainly of interest at the generic level but are also related to tribal divisions and may be of phylogenetic significance in the family. Other classes of flavonoid are rare: leucocyanidin was detected once in Apiastrum, and the glucoxanthone mangiferin once in Heptaptera. Furanocoumarins were found in the leaves mainly of Angelica, Peucedanum and Seseli species but a survey of seeds of 130 species showed that these compounds were widespread in the family, some correlation with tribal divisions being apparent. Examination of the Umbelliferae for presence of polyacetylenes, simple hydroxycoumarins and the rare sugars, apiose and umbelliferose, has shown that these substances are widespread and consequently of little systematic interest within the family. Soluble proteins and the enzymes peroxidase and esterase present in the seed of selected species from all tribes in the Apioideae were studied by acrylamide gel electrophoresis. Distinct differences in patterns were found to be present at the tribal and generic levels. In some cases, the macromolecular supported the micromolecular data in conforming generic separations. The general value of the various chemical characters in the systematics of the family is discussed.
Extraction of maturing Phaseolus vulgaris seeds with an ascorbic acid—NaCI medium facilitated the preparation of two globulin fractions which wer
DNA (deoxyribonucleic acid) base composition of 61 species belonging to 35 families of angiosperms, both dicotyledons and monocotyledons, were determined with a view to assessing its taxonomic and phylogenetic significance. The data indicated a considerable similarity amongst the forms investigated, the range of GC content being about 13%. When the dicotyledons were considered separately, the range was 6%. Though the variation was found to be statistically significant, no clear cut delimitation was discernible at any taxonomic level. The results are discussed in the light of recent researches on plant DNA.
DNA was isolated from three genera of ferns (Angiopteris, Ophioglossum, Ciboteum) and three genera of fern-allies (Psilotum, Equisetum, Selaginella), using a homogenization medium designed to minimize action of phenoloxidases.The DNA base compositions of these evolutionarily diverse plants fell within the narrow range of 37–41 % (G+C) as determined by buoyant density in CsCl, except for Selaginella emiliana and S. kraussiana which were 45 and 46 % (G+C). The latter also contained satellite DNA's. The overall range of % (G+C) was comparable to that of angiosperms. This suggests that the 400 million years since the time of divergence of land plants has been too short for large changes in the DNA base composition to have occurred.
The pollen of cocksfoot grass (Dactylis glomerata) is an important cause of allergic reactions in man. Preliminary studies, which established that constituents of an extract of this pollen could be separated, and then recovered efficiently, by size-exclusion chromatography on TSK G3000 SW, have been extended. A comparative examination has been made by this procedure of cocksfoot pollen extracts from different sources and of several batches of extract from one source. The recovery and distribution of biological activity has been assessed by the radioallergosorbent test, and the results have been used for the selection of fractions for further investigation by chromatography and electrophoresis. Two constituents active in the radioallergosorbent test have been purified from the extracts.