Aluminum Chloride Colorimetric Assay Research Articles

Effects of LED treatments on the growth and nutritional content of lettuce (Lactuca sativa) in a hydroponic vertical farming system

Introduction: LED-integrated hydroponic vertical farming system (HVFS) may address food security challenges arising from urbanisation, while antioxidants in vegetables may prevent cardiovascular disease. This study aimed to explore the LED effects on yield, antioxidant properties, and vitamin C content in Butterhead lettuce (BL) and Italian lettuce (IL). Methods: Three LED combinations were used: 77% red, 14% green, and 9% blue lights (77R:14G:9B); 7% red, 41% green, and 52% blue lights (7R:41G:52B); and 25% red, 67% green, and 8% blue lights (25R:67G:8B). The concentration and pH of nutrient solution in HVFS ranged from 0.878-2.061ppm and 6.0-6.6, respectively, with a 12:12 light-dark cycle. Chlorophyll and anthocyanin contents were measured using chlorophyll and anthocyanin meters, respectively. Total phenolic and vitamin C content were measured using Folin-Ciocalteu colorimetric assay. Total flavonoid and antioxidant activity were determined through aluminium chloride colorimetric assay and 2,2-diphenyl-1-picrylhydrazyl (DPPH) colorimetric assay, respectively. Completely randomised design was adapted with three replications. Results: BL exhibited greater yield, anthocyanin and chlorophyll contents, while IL had higher vitamin C content. Significantly (p<0.001) higher yields were produced under 77R:14G:9B and 25R:67G:8B. Significantly higher phenolic (p<0.001) and flavonoid (p=0.003) contents were produced under 25R:67G:8B. Vitamin C was significantly (p=0.037) higher under 25R:67G:8B than 7R:41G:52B. Conclusion: LED treatment with higher proportion of red light generated greater yield, whereas higher proportion of green light gave higher phenolic, flavonoids, and vitamin C accumulation. This study provided preliminary data on boosting crop yield and phytochemical contents, which may improve food self-production and population health.

In Silico Antibacterial Activity of Polyacetilene Derivatives Against Mycobacterium tuberculosis and In-Vitro Antioxidant Properties from Ethanol Extraction of Blackjack (Bidens pilosa L.)

Blackjack (Bidens pilosa L.) has a bioactive compound, one of which is polyacetylene, which can inhibit the growth of antimicrobials in general. This study aims to conduct an in silico study to determine the antibacterial activity against Mycobacterium Tuberculosis of 20 derivatives polyacetylene; TPC was determined by Folin–Ciocalteu colorimetric method using gallic acid as standard, and various concentrations of the extract solutions were measured at 741 nm. TFC was calculated using an aluminum chloride colorimetric assay. Quercetin was used as standard, and the absorbance was measured at 426 nm. Antioxidant activity was evaluated using DPPH scavenging 517 nm and Frap Assay 596 nm. The molecular docking results showed that the compounds (5-(2-Phenylethynyl)-2-b-glucosylmethyl-thiophene, Glucopyranosyloxy-3-hydroxy-6(E)-tetradecane-8,10,12-triyne and Phenylhexa-1,3,5-triyn-1-yl acetate were most potentially anti-tuberculosis on the target protein InhA with the binding energy produced <-6.55 Kcal/mol and the lowest inhibitory cost of 1.838×10-6. The compound has amino acids similar to natural ligands and standard compounds of isoniazid. The given ADME-T predictions showed good pharmacokinetic properties and safe levels of toxicity (5-(2-Phenylethynyl)-2-b-glucosylmethyl-thiophene, Glucopyranosyloxy-3-hydroxy-6(E)-tetradecane-8,10,12-triyne in hepatotoxic and LD50. The highest antioxidant properties in blackjack are produced by FRAP assay 722.905±7.985 mMTE/gr. This proves that blackjack has antioxidant activity and is required for bioassay-guided isolation testing against Mycobacterium tuberculosis.

Comparison of the Bioactive Properties of Honey Proteins from Floral Sources in Indonesia

The diversity of plant species in Indonesia has resulted in a wide variety of floral honey. The antioxidant activities of these Indonesian honeys have not been determined. Ten types of honey from indigenous Indonesian sources (9 monofloral; 1 heterofloral) were evaluated for their antioxidant activity using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method. The Folin-Ciocalteu test was used to determine the total phenolic content, and an aluminum chloride colorimetric assay was used to determine the total flavonoid content. SDS–PAGE was used to determine the molecular weights of the honey proteins. The mean antioxidant activity of the honey samples, defined as the IC50 of DPPH, ranged from 5.46 to 34.42. The mean total phenol content ranged from 27.57 to 53.85 mg GAE/100 g, and the mean flavonoid content ranged from 3.58–15.67 mg CE/100 g. A protein of ~60 kDa was detected in all honey varieties. A positive correlation was observed between antioxidant activity, total phenols, and total flavonoids, indicating that their phenolic and flavonoid components confer the antioxidant activity of these honey varieties. To the best of our knowledge, indigenous Indonesian honey, especially monofloral Calliandra honey, has shown greater antioxidant activity than other kinds of honey worldwide.

Correlation of Antioxidant Properties Between Immature and Mature Okra (Abelmoschus Esculentus) Fruits

This study aimed to assess and compare the antioxidant activity and content (total flavonoid levels and total phenolic) of mature and immature okra. The antioxidant activity of okra fruits was assayed using four methods, namely: Aluminium Chloride Colorimetric assay, Folin-Ciocalteu assay, 1, 1-diphenyl-2-picrylhydrazyl (DPPH), and Reducing Antioxidant Power assay (FRAP) assays. The immature, mature, and very mature okra samples (less than 8 days, 10-15 days, and more than 20 days, respectively) were extracted using two different solvents (65% ethanol and water). The sample that was extracted with mature ethanol had the highest Total Phenolic Content (TPC) at 21.564 ± 1.635 mg GAE/g, while the sample that was extracted with extremely mature ethanol had the highest TFC at 54.391 ± 8.224 mg QE/g. The mature 65% ethanolic extracted sample showed the lowest IC50 value of DPPH scavenging activity (0.920± 0.096 mg/ml), and the mature ethanol extracted sample had the highest FRAP value (232.018± 5.337 μmol Fe2+/g). These studies showed that ethanolic extracts of mature Abelmoschus esculentus had higher antioxidant content and activity than okra water extracts. Based on the DPPH Radical Scavenging Assay revealed favourable associations between TPC (r = 0.860), TFC (r = 0.742), and antioxidant activity as evaluated by FRAP, demonstrating that both phenolics and flavonoids contributed to the extract’s antioxidant properties. Both TPC and TFC showed negative correlations with IC50 values (r = -0.766, r = -0.650, respectively). In conclusion, the mature okra fruits extracted with 65% ethanol give higher antioxidant content than the water extracts of okra fruits and potentially be used as a source of antioxidants rather than be discarded.

Phytochemical profiling and bioactivity study of Adenia panduriformis in Zambia using UHPLC-MS/MS-MZmine3, GNPS, and METLIN Gen2

Diseases, especially degenerative illnesses, can be treated and avoided altogether, and good health can be enhanced by including health-promoting foods in the diet without side effects. Adenia panduriformis (Passifloraceae) is a wild vegetable consumed as part of the everyday diet in some communities in Zambia. The plant is also used in traditional medicine to effectively treat and manage many diseases, including degenerative diseases. Because of significant health benefits and medicinal attributes, this vegetable has very high merit to be investigated for its phytochemicals and bioactivities. Therefore, this study screened phytochemicals using qualitative chemical tests and Liquid chromatography-tandem mass spectrometry (LCMS/MS), identified metabolites using molecular networking tools and evaluated the total polyphenolic content and antioxidant activity of the methanolic leaf extract of the vegetable. Solvent extraction by maceration recovered metabolites for screening by qualitative chemical tests, extraction by QuEChERS (quick, easy, cheap, effective, rugged, and safe) was used to recover metabolites for screening by LCMS/MS, and tentative phytochemical constituents were identified using MZmine3, GNPS (global natural products social molecular networking) and METLIN. Total phenolic content (TPC) determination used the Folin-Ciocalteu (FC) method at 765 nm. Total flavonoid content (TFC) was evaluated using the aluminium chloride colorimetric assay at 510 nm. Antioxidant activity was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay at 517 nm. Phytochemical screening by qualitative chemical tests revealed the presence of many classes of phytocompounds, such as alkaloids, polyphenols, terpenoids, and saponins. TPC was determined to be 122.30 ± 2.00 mg GAE/g, and TFC was 60.17 ± 0.25 mg QE/g. Screening by LCMS/MS revealed the presence of many metabolites, of which 23 phytocompounds were identified using MZmine3, 42 by GNPS, and 21 by METLIN. The crude extract exhibited high DPPH free radical quenching activity with a relatively low IC50 value of 53.48 µg/mL compared to the reference standard ascorbic acid IC50 of 74.47 µg/mL. The significantly lower IC50 value for the wild vegetable than that of ascorbic acid indicates a considerably high efficiency of the metabolites in the vegetable as antioxidants. The presence of many metabolites with health-promoting properties and high antioxidant activity of the leaf extract of A. panduriformis may be the science behind its remarkable health benefits and power to treat and manage a variety of diseases, including degenerative illnesses. This study presents results that indicate the potential of A. panduriformis as a rich source of metabolites with brilliant health-promoting properties and high antioxidant activity, which may open exciting opportunities for further development in the fields of pharmacology and drug development.

THE STUDY of SOME BIOLOGICAL ACTIVITIES of CUSCUTA CHINENSIS and ITS TWO HOSTS

Introduction: Cuscuta, (Ces as Persian name) is a parasitic plant with different biological effects and applications. The metabolic profile and effects of the Ces depend on the host on which it grows. Its main components are phenols and flavonoids. Considering the antioxidant effects of these compounds, the purpose of this research was to investigate the antioxidant activities and total phenolic and flavonoid contents of Cuscuta as well as Mellissa officinalis and Alhaji murarum as hosts.
 Methods: Aqueous, hydroalcoholic extracts of the Cuscuta grown on two hosts were prepared separately. Hydroalcoholic extracts of host plants were also prepared. Total phenolic and flavonoid contents were evaluated using Folin- Ciocalteu reagent and aluminum chloride colorimetric assays, respectively. In order to investigate the antioxidant activity, the ability of extracts to scavenge DPPH free radical (DPPH 2, 2-(diphenyl-1-picrylhydrazyl) and their capacity to reduce iron 3+ion were measured by FRAP method.
 Results: The results showed that all the examined extracts have significant phenolic and flavonoid contents. The highest antioxidant activity was observed in hydroalcoholic extracts, which was proportional to the amount of phenolic and flavonoid contents in the extracts. In addition, the extracts of host plants were richer than the parasite extracts. The extract of Mellissa officinalis showed better effects than Alhaji murarum in all of four investigated factors.
 Conclusion: According to the findings of the present study, the hydroalcoholic extracts of the Cuscuta plants are rich in phenolic and flavonoid compounds, which may be the cause of the antioxidant activity. Of course, the compounds and effects of Cuscuta depend on the host. In future studies, the plant can be grown in different hosts.

Phytochemical screening, TLC profile and antioxidant activity of hydromethanolic extract of Centaurothamnus maximus leaves

The species Centaurothamnus maximus (Forssk.) Wagenitz & Dittrich is grown in different places in Yemen and is used to treat many diseases. The aim of this research was to investigate the phytochemicals and the antioxidant activity of hydromethanolic extract obtained from this plant. For preliminary phytochemical analysis, standard procedures were applied. The Folin–Ciocalteu method was used to evaluate the total phenolic acid content of the plant extract, while the total flavonoid content was determined using the aluminum chloride colorimetric assay. The antioxidant activities were assessed using the 1,1-diphenyl-2-picrylhydrazyl. Chemical analysis revealed the presence of carbohydrates, saponins, polyphenols, flavonoids, triterpenes, sterols, tannins and alkaloid. The total content of phenols was 43.04±3.30 mg gallic acid equivalent per gram dry extract, while the total flavonoid content was 36.60±2.86 mg rutin equivalent per gram dry extract. With TLC and specific reagents, flavonoid and phenolic substances were identified. The hydromethanolic extract and its fractions showed a dose dependent scavenging activity, we found that the ethyl acetate fraction has high inhibitory percentages equivalent to IC50 40.86±0.001 for DPPH. These results indicate that the leaves of Centaurothamnus maximus are rich in secondary metabolites, and may be used as a natural anti-oxidant source. Extensive chemical and pharmacological studies should be carried out

A comparative study of Ghanaian propolis extracts: Chemometric analysis of the chromatographic profile, antioxidant, and hypoglycemic potential and identification of active constituents

Diabetes is a disease characterized by high post-prandial glucose levels, which lead to other complications such as peripheral end organ damage. The use of enzyme inhibitors in the management of Type-2 diabetes ensure the control of blood glucose levels via the control of carbohydrate metabolism. The use of standard agents such as acarbose is associated with unwanted side effects hence the need to investigate other sources of antihyperglycemic agents. Propolis, a natural substance from bees, possesses diverse biological activities including antioxidant, antimicrobial and antidiabetic properties. However, the phytochemical content of propolis and its extracts may vary depending on the geographical area, the solvent of extraction and type of bees. This study represents the first attempt to compare different extracts of propolis from the same source in sub-Saharan Africa. In this study, the effect of solvent and source of Ghanaian propolis on parameters such as the total phenolic and flavonoid contents, chromatographic profile, antioxidant and α-amylase inhibitory effects were investigated with the aim of identifying and characterizing the most promising extract, which could be of direct or indirect benefit in the management of Type-2 diabetes.Combinations of water, ethanol-water and ethanol extracts were prepared from propolis from three regions. Phytochemical screening was performed on the extracts after which the Folin Ciocalteu method and aluminum chloride colorimetric assay were used to estimate the total phenolic and flavonoid contents respectively. Antioxidant potential of extracts was estimated using DPPH and phosphomolybdenum assays. In-vitro α-amylase inhibition assay was used to investigate hypoglycemic effect of the extracts. Statistical tools such as ANOVA, principal component analysis, hierarchical cluster analysis employed to determine sources of variations within the data obtained, to classify the extracts based on activity and to predict the most effective extract. This extract was then subjected to UHPLC-Q-TOF MS/MS and GC–MS techniques to characterize the constituents.Chemometric analysis of the data obtained showed that the variations in the data could be explained by both propolis source and extraction solvent. Though ethanol extracts generally contained more constituents, the more notable activities were in the ethanol-water extracts. The ethanol-water extract of Bono East propolis (EWBE) was the most potent DPPH radical scavenger (IC50 of 149.37 ± 2.90 µg/mL as compared to 116.60 ± 0.93 µg/mL GAE standard). It was also one of the three extracts which were more potent than acarbose (369.89 µg/mL) in the α-amylase inhibition assay. The predominant constituents from the LC-MS dereplication of EWBE were caffeic acid and flavonoid derivatives whilst 5,5-dimethyl-1-oxa-5 silacyclononanone-9 was the most significant active constituent identified through the GC–MS analysis. The identified constituents are known to have strong antioxidant and antidiabetic properties.The effects of source and solvent of extraction on the biological and physicochemical properties of propolis in Ghana have been quantified using statistical tools. The combined biological effects of propolis suggest a possible role in their usage in the management of type-2-diabetes and its related complications. Ethanol-water extracts were the most promising with EWBE showing the strongest antihyperglycemic activity. Such extracts represent leads towards further research into toxicity and formulation in order to develop safe and useful products for the management of type-2 diabetes.

Evaluation of Nootropic Potential of Aerva persica Roots against D-galactose-induced Memory Impairment.

The primary phytoconstituents reported to have neuroprotective effects are flavonoids and phenolic compounds. Aerva persica roots are reported to be rich in flavonoids and phenolic compounds. Therefore, this study aimed to explore the nootropic potential of Aerva persica roots. The objective of this study was to evaluate the nootropic potential of Aerva persica roots against D-galactose-induced memory impairment. In this study, the roots of Aerva persica were extracted with 70% ethanol. The obtained extract was evaluated for total phenolic content using the Folin-Ciocalteu method and total flavonoid content using the aluminium chloride colorimetric assay. Afterward, the acute oral toxicity of the extract was determined following the Organisation for Economic Co-operation and Development (OECD) guideline 423. Additionally, two doses of Aerva persica (100 and 200 mg/kg body weight (BW)) were evaluated for their nootropic potential against D-galactose-induced memory impairment. The nootropic potential of the crude extract was assessed through a behavioural study and brain neurochemical analysis. Behavioural studies involved the evaluation of spatial reference- working memory using the radial arm maze test and the Y-maze test. Neurochemical analysis was performed to determine the brain's acetylcholine, acetylcholinesterase, glutathione (GSH), and malondialdehyde (MDA) levels. The total phenolic content and total flavonoid content were found to be 179.14 ± 2.08 μg GAE/mg and 273.72 ± 3.94 μg QE/mg, respectively. The Aerva persica extract was found to be safe up to 2000 mg/kg BW. Following the safety assessment, the experimental mice received various treatments for 14 days. The behavioural analysis using the radial maze test showed that the extract at both doses significantly improved spatial reference-working memory and reduced the number of total errors compared to disease control groups. Similarly, in the Y-maze test, both doses significantly increased the alteration percentage and the percentage of novel arm entry (both indicative of intact spatial memory) compared to disease control. In neurochemical analysis, Aerva persica at 200 mg/kg significantly normalised the acetylcholine level (p<0.0001) and GSH level (p<0.01) compared to disease control. However, the same effect was not observed with Aerva persica at 100 mg/kg. Additionally, Aerva persica at 200mg/kg BW significantly decreased the acetylcholinesterase level (p<0.0001) and decreased the brain's MDA level (p<0.01) compared to the disease control, whereas the effect of Aerva persica at 100 mg/kg BW in reducing acetylcholinesterase was non-significant. Based on the results, it can be concluded that the nootropic potential of Aerva persica was comparable to that of the standard drug, Donepezil, and the effect might be attributed to the higher content of flavonoids and phenolic compounds.

Comparison of Physicochemical Properties of Biscuits Supplemented With Soy, Kinema, and Buckwheat Flour

Consumer demand for bakery products prepared using composite flour, such as biscuits, is growing. The incorporation of buckwheat flour, soy flour, and kinema flour can be considered in composite flour-based biscuits since they have good nutraceutical properties and their gluten-free nature can help avoid celiac disease. The purpose of this research was to study the physicochemical and functional characteristics of buckwheat flour, soy flour, and kinema flour as well as their potential as functional foods. The biscuits were prepared with a mixture of buckwheat flour, kinema flour, and soy flour in 10%, 30%, and 50% concentrations with whole wheat flour to evaluate the quality and acceptability of the biscuit. The composite flour biscuits were examined for their proximate compositions. The DPPH radical scavenging assay was used to assess the antioxidant properties of the biscuit extract, while the Folin-Ciocalteu method and the Aluminum Chloride Colorimetric Assay were both used to assess the total phenolic and flavonoid contents, respectively. The overall acceptability of biscuits by sensory analysis was highest at a 50% level of Black kinema flour (BLKF). Sample M (BLKF 50 %) had 0.66, 21.06, 1.61, 11.88, and 3.23 % moisture, fat, crude fiber, protein, and ash, respectively. The highest total phenolic and total flavonoid contents were found in black kinema flour (748.22 mg GAE/g and 8.92 mg GAE/g, respectively), while the highest level of antioxidant activity was found in brown kinema flour (313.56 μg/ml). BRKF 50:50 had the highest antioxidant activity (440.43 μg/ml), while BLKF 50:50 had the highest total phenolic and flavonoid contents (692.67 mg GAE/g and 8.60 mg GAE/g, respectively). The qualitative evaluation of the phytochemical content and amino acid in various flour varieties as well as in composite biscuits gave positive results. Thus, the composite flour biscuits could serve as a low-cost functional food.

TOTAL FLAVONOID CONTENT AND ANTIOXIDANT ACTIVITYBY DIFFERENT DRYING AND EXTRACTION METHODS OF Clinacanthus nutans LEAVES

This report presents a study on the total flavonoid content and antioxidant activity of Clinacanthus nutans leaves from different drying and extraction methods. The C. nutans leaves were subjected through three different drying methods: oven (40°C, 60°C and 80°C), vacuum oven and air drying. Afterwards, extraction on the dried leaves was performed using three different extraction techniques: soxhlet, maceration and ultrasoundassisted solvent extraction. Finally, total flavonoid and antioxidantactivity was determined spectrophotometrically by aluminium chloride colorimetric assay and 2-Diphenyl-1-picryl hydrazyl (DPPH) method respectively. The results showed that the choice of drying and extraction methods had a significant influence on total flavonoid and antioxidant content of C. nutans extract. The combination of drying C. nutans leaves using laboratory oven at 60°C and soxhlet extraction obtained the highest amount of total flavonoid and antioxidant content at 24.53 ± 0.95 mg RU/g and 89.73 ± 4.39mg TE/ g respectively. The highest antioxidant activity for C. nutansfromair-dried samples and vacuum oven-dried samples were obtained through maceration extraction at 85.46 mg TE/ g and 83.96 mg TE/ g respectively.

UHPLC-MS/MS phytochemical screening, polyphenolic content and antioxidant potential of Diplorhynchus condylocarpon (Müll.Arg.) Pichon (Apocynaceae), a medicinal plant

Diplorhynchus condylocarpon (Müll.Arg.) Pichon (Apocynaceae) leaves, stem bark, and roots are commonly used as popular folk medicine in Zambia and other African countries. This work aimed to find out the broad categories of phytocompounds present in the plant, screen the polyphenols, and assess the total polyphenolic content and the antioxidant activity of methanolic extracts of leaf, stem bark, and roots. Maceration was used to extract secondary metabolites for preliminary phytochemical screening. QuEChERS (quick, easy, cheap, effective, rugged, and safe), based technique was used to recover polyphenols for liquid chromatography with tandem mass spectrometry (LC-MS/MS) screening. Standard chemical tests were used for preliminary phytochemical screening. Total phenolic content (TPC) was evaluated using the Folin-Ciocalteu (FC) method at 765 nm. Total flavonoid content (TFC) was evaluated by the aluminum chloride colorimetric assay at 510 nm. Antioxidant activity was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method at 517 nm. The preliminary phytochemical screening revealed the presence of antioxidants, polyphenols, and other valuable phytocompounds such as alkaloids, saponins, and terpenoids. The LCMS/MS polyphenolic screening revealed the presence of mainly polyphenolic metabolites. TPC values were higher in the leaf (171.099±0.515 mg GAE/g) and root (88.8093±3.156 mg GAE/g) than in the stem bark (71.061±0.515 mg GAE/g) extracts. Further, the leaf extract contained 129.402±0.407, the root contained 64.672±0.813, and the stem bark had 37.643±0.704 mg QE/g total flavonoids. The leaf extract exhibited in a dose-dependent manner higher DPPH radical scavenging activities (34.250, 50.625, 66.562, 81.406, and 97.344%), followed by the root (16.406, 31.406, 50.781, 65.625, and 82.188%), and the stem bark extract only had the least DPP quenching activities (15.469, 27.969, 40.469, 52.500, and 68.590%) for the concentration of 20, 40, 60, 80 and 100 µg/mL respectively. Further, the results from the DPPH assay show that the leaf extract had the best IC50 value of 19.973 µg/mL, followed by the root extract with 60.867 µg/mL, and the stem bark extract with 107.150 µg/mL. These antioxidant activities support the traditionally claimed therapeutic benefits of D. condylocarpon leaves, stem bark, and roots in the treatment of oxidative stress-related disorders such as diabetes mellitus, chronic wounds, and diarrhea. This study may lead to the production of new herbal medicines using D. condylocarpon.

Comparison of the total phenolic and flavonoid contents of Amomum compactum Sol. Ex Maton from districts Linggo Asri and Paninggaran, Pekalongan Regency

Cardamom (Amomum compactum Sol. Ex Maton) is one of the largest contribution in biopharmaceutical production plants in Indonesia that is equal to 12.22% and yield reaching 105.735 kg/year in Pekalongan Regency. The largest cardamom production in Pekalongan Regency is in 2 districts, namely Linggo Asri and Paninggaran. Cardamom is used in traditional medicines, because it has chemical compounds such as flavonoids and phenolics. The aim of this study was to compare total flavonoids and total phenolics content in Amomum compactum Sol. Ex Maton extract originates from Linggo Asri and Paninggaran district, Pekalongan Regency. The samples were extracted by water extraction, and then phytochemical contents were identified with HCl and FeCl3 reagent. The samples were calculated of total flavonoids and total phenolics content by spectrometric using aluminum chloride colorimetric assay and Folin–Ciocalteu method. The result showed a significant difference in the levels of total phenolic content between Amomum compactum Sol. Ex Maton extract origin from Linggo Asri district 41.43±0.54 µg/mg and Paninggaran district 120.55±1.89 µg/mg, but it was not found in total flavonoid content where from Linggo Asri district 289.06±4.1 µg/mg and Paninggaran district 303.81±0.5 µg/mg. The conclusion is Amomum compactum Sol. Ex Maton extract originates from Linggo Asri and Paninggaran district of Pekalongan city showed a significant difference in the levels of total phenolic content, but it was not found in total flavonoid content.

Phytochemical constituents, antioxidant and antibacterial activities of Plectocephalus varians (A. Rich.) C. Jeffrey ex Cufod root extracts

Plants have been used to treat diverse types of diseases in different cultural groups around the globe. In this regard, the root of Plectocephalus varians (P. varians) is claimed to have a beneficiary effect in treating cancer and hemorrhoids in Ethiopia. Hence, this study aimed at the phytochemical investigation, antioxidant, and antibacterial activities of n-hexane, acetone, and methanolic extracts of P. varians root. The different crude extracts of P. varians were obtained through maceration technique. The total phenolic content (TPC) and total flavonoid contents (TFC) of the extracts were estimated using Folin-Ciocalteu Reagent (FCR) and aluminum chloride colorimetric assays, respectively. The antioxidant activities of the extracts were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and ferric reducing antioxidant power (FRAP) assays. The antibacterial activities of the extracts were assessed by using disc diffusion method. The results echoed the presence of alkaloids, saponins, flavonoids, steroids, phenols, tannins, anthraquinones, terpenoids, polyphenols, and glycosides in the root of P. varians. The methanolic root extract (MRE) had the highest TPC (107.18 mg GAE/g) and TFC (120.194 mg QE/g) followed by acetone root extract (ARE) (TPC = 98.68 mg GAE/g; TFC = 64.038 mg QE/g) and n-hexane root extract (HRE) (TPC = 12.39 mg GAE/g; TFC = 9.917 mg QE/g). The DPPH radical scavenging and FRAP assays demonstrated the antioxidant effects of HRE (IC50 = 681.75 ppm; EC50 = 60.65 ppm), ARE (IC50 = 165.73 ppm; EC50 = 51.67 ppm) and MRE (IC50 = 132.06 ppm; EC50 = 30.97 ppm) of P. varians. Furthermore, the root fractions elicited pronounced dose-dependent growth inhibition against Staphylococcus aureus, Streptococci pyogenes, Escherichia coli, and Klebsiella pneumoniae with mean zone of inhibition (MZI) ranging from 11 ± 0.38 to 20 ± 0.04 mm at 800 ppm. Overall, the present study provides ethnopharmacological evidence suggesting the medicinal importance of P. varians. The results also call for further bioassay-guided phytochemical screening and in vitro and/or in vivo bioactivity testing.

In vitro anti-ageing activities of ethanolic extracts from Pink rambutan (Nephelium lappaceum Linn.) for skin applications

Skin ageing is characterized by features such as wrinkles, loss of elasticity, laxity, rough-textured appearance, melasma and freckles. Several researches have focused for preventing, and treating skin ageing by many natural ingredients. This study aimed to assess the anti-ageing activities for anti-skin ageing of the ethanolic extracts of Pink rambutan (PR) (Nephelium lappaceum Linn.) from leaves (L), branches (B), seeds (S), and peels from ripe (R) and young (Y) fruits. The extraction yields of all Pink Rambutan (PR) extracted by the Maceration (M) and the Soxhlet extraction (Sox) using 95% ethanol as a solvent, ranged from 10.62% to 30.63%. Flavonoids were found as the main phytochemicals in almost all the PR extracts. The PR-Y-M and PR-Y-Sox extracts gave the highest total phenolic contents by the Folin-Ciocalteu assay of 67.60 ± 4.38 mgGAE/g, and total flavonoid contents by the modified aluminum chloride colorimetric assay of 678.72 ± 23.59 mgQE/g, respectively. The PR-L-M extracts showed the highest three anti-oxidative activities; the free radical scavenging (SC50 of 0.320 ± 0.070 mg/mL), the lipid peroxidation inhibition (LC50 of 0.274 ± 0.029 mg/mL), and the metal chelation activity (MC50 of 0.203 ± 0.021 mg/mL). All the PR extracts at 0.01 and 0.1 mg/mL showed no cytotoxicity on B16F10 cells, and human skin fibroblasts, respectively. Likewise, the PR-R-Sox extract exhibited the highest anti-melanogenesis on B16F10 cells (52.7 ± 0.9%) and, the mushroom tyrosinase inhibition activity (IC50 of 0.04 ± 0.02 mg/mL), which was significantly comparable to kojic acid (p < 0.05). The PR-Y-Sox extract showed the collagen biosynthesis by the Sirius Red method, and the stimulation of anti-ageing genes (Sirt1 and Foxo1) on human skin fibroblasts by the RT-PCR method, which were similar to standards ʟ-ascorbic acid and resveratrol, respectively. This study suggests that the PR-R-Sox and PR-Y-Sox extracts can be further developed as natural anti-ageing agents for whitening and anti-wrinkle in the cosmetics, cosmeceutical, and pharmaceutical industries.

Analytical determination, antioxidant and anti-inflammatory activities of Bhamrung-Lohit a traditional Thai medicine.

Bhamrung-Lohit (BRL) remedy is a traditional Thai medicine (TTM). There are few reports of biological activity, the activity of its constituent plants, or quantitative analytical methods for the content of phytochemicals. In this study, we investigated antioxidant, anti-inflammatory activity, and total phenolic and flavonoid content and validated a new analytical method for BRL. Antioxidant activity was evaluated by a 2,2-diphenyl-1-picrylhydrazyl (DPPH and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging. The cellular antioxidant activity was evaluated by inhibition of the superoxide anion (O2●-) production from HL-60 cells and anti-inflammatory activity by inhibition of nitric oxide production in RAW264.7 cells. The total phenolic and flavonoid contents were analyzed using the Folin-Ciocalteu method and an aluminum chloride colorimetric assay, respectively. Validated analytical procedures were conducted according to International Conference on Harmonization (ICH) guidelines. An ethanolic extract of BRL exerted potent DPPH radical scavenging activity and moderate antioxidant and anti-inflammatory activity. Caesalpinia sappan exerted the greatest effect and the highest content of total phenolics and flavonoids. The HPLC method validated parameters that complied with ICH requirements. Each peak showed selectivity with a baseline resolution of 2.0 and precision was less than 2.0% CV. The linearity of all compounds was > 0.999 and the recovery % was within 98.0%-102.0%. The validated results demonstrated specificity/selectivity, linearity, precision, and accuracy with appropriate LOD and LOQ. BRL remedy, a TTM demonstrated antioxidant and anti-inflammatory properties. This study is the first report on the biological activity and the validation of an HPLC method for BRL remedy.

GC-MS Profiling and Bioactivities of Essential Oil and Extracts of Cinnamomum tamala (Buch.-Ham.) Nees &amp; Eberm. Leaves from Kathmandu Valley, Nepal

The present work reports the phytoconstituents present in the essential oil and four different solvent extracts of Cinnamomum tamala leaves. Their antibacterial, antifungal, and antioxidant potential were also evaluated. The extraction of essential oil was performed by hydro-distillation using Clevenger apparatus and its chemical composition was identified by gas chromatography coupled with mass spectrometry (GC-MS). The leaf extracts were obtained by the cold percolation method. Linalool and cinnamaldehyde (E) were major compounds in the oil. Phytochemical screening of the extracts revealed the presence of terpenoid, glycoside, tannin, reducing sugar, polyphenol, saponin, flavonoid, and alkaloid. Total phenolic content (TPC) and total flavonoid content (TFC) were quantified using Folin-Ciocalteu and Aluminium chloride colorimetric assay, respectively. The methanol extract (98.36 mg GAE/g) and ethyl acetate extract (90.44 mg GAE/g) showed higher TPC values. Similarly, the TFC value of ethyl acetate extract was higher (478.78 mg QE/ g) than the other extracts. The antioxidant activity of extracts was assessed by using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay in which the ethyl acetate extract showed high antioxidant efficacy. The essential oil and chloroform extract showed antifungal activity against Candida albicans, while only the oil showed activity against Shigella dysenteriae.

A Comparative Study on Ascorbic Acid Concentration, Total Phenol, and Flavonoid Content in Citrus Species Grown in a Different Region of Western Nepal.

Different genetic and environmental factors like altitude, temperature, light radiation, etc. affect the production of phytoconstituents like ascorbic acid, phenol, flavonoid, tannin, etc. So, it is necessary to evaluate phytoconstituents quantitatively and qualitatively because of their different medicinal values. This study is aimed at evaluating and comparing ascorbic acid concentration, total phenol, and flavonoid content in citrus species grown in different places in Western Nepal. For this, fruit collected from Lamjung and Nawalparasi was peeled, and collected juice was extracted with ethanol. Ascorbic acid was estimated using titration with dichlorophenolindophenol dye, and total phenol was estimated using Folin reagent; whereas, flavonoid content was determined by aluminum chloride colorimetric assay. The result showed the maximum ascorbic acid concentration in the fruit juice of Citrus maxima collected from Lamjung, i.e., 2.98 ± 0.161 mg/100 ml, and the least concentration was recorded on extract of Citrus limon collected from Nawalparasi, i.e., 1.005 ± 0.205 mg/100 ml. The total phenol content was comparatively higher in Citrus maxima and Citrus aurantifolia collected from Lamjung, i.e., 12.48 ± 0.40 mg GAE/gm, respectively. Flavonoid content was comparatively higher in Citrus maxima collected from Nawalparasi, i.e., 484.466 ± 3.055 mg QE/gm. This study showed a variation in the concentration of chemical constituents between different places in Western Nepal. This might be due to differences in climatic conditions, environmental factors, altitude, temperature, etc. This study helps to reflect the best-suited altitude for commercial cultivation of the citrus species as these phytochemicals have different medicinal values. Also, this study can help the food industry to use an alternative source of synthetic antioxidant such as ascorbic acid, phenol, and flavonoid. And local people will economically benefit by the commercial cultivation of citrus fruits.

Effect of Extraction Techniques on Yield, Total Phenolic Content, Total Flavonoid Content and its Antioxidant Activity of Leaves, Stems and Roots from Cassia alata Linn

Cassia alata Linn. (Senna alata) or commonly known as “gelenggang” in Malaysia, has been reported to have therapeutic properties and introduced as a resource for traditional remedies for thousand years. Its therapeutic properties were including antibacterial, antifungal, antioxidant, anthelmintic, antilipogenic, antidiabetic, dermatophytic, and anti-hyperlipidemic [1].&#x0D; &#x0D; In this study, the use of soxhlet extraction and ultrasonic assisted extraction for different parts of C. alata were investigated. For each extraction techniques, the amount of sample, volume of solvent extraction and extraction period were different. Hexane was used as the extraction solvent. The total phenolic content (TPC), total flavonoid content (TFC), and its antioxidant activities were evaluated using Folin-Ciocalteu’s method, aluminum chloride colorimetric assay, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, respectively. &#x0D; &#x0D; The experimental results showed the highest yield of extract in leaves (4.86±0.24%), stems (0.75±0.09%) and roots (0.59±0.01%) were achieved by using soxhlet extraction technique. The results of TPC and TFC were varied from 0.049±0.004 mg GAE/g DW to 0.185±0.004 mg GAE/g DW and 0.023±0.001 mg QE/g DW to 0.203±0.007 mg QE/g DW, respectively. The highest were recorded from the soxhlet extraction for both of TPC and TFC results. Meanwhile, the radical scavenging activity was the highest from the roots of C. alata, by soxhlet extraction at 21.663±2.097% (60 µg/ml), with the IC50 value at 9.96 µg/ml (Table 1).&#x0D; These findings suggest that the selection of soxhlet extraction as a technique to extract C. alata using hexane was more effective in comparison to ultrasonic assisted extraction. For the further references on the potential of this technique, it is suggested that this study is further analyzed for the identification of its bio-active compounds.

Assessment on antioxidant activity of the aqueous leaf extracts of Combretum microphyllum and the effect of Co(II)-leaf extract complex on antibacterial activity of leaf extracts of the plant material

BackgroundIn the present study, different aqueous leaf extracts of C.microphyllum prepared by maceration were investigated for their antioxidant potential, antibacterial activity, and polyphenolic and flavonoid contents. MethodLeaf samples of C. microphyllum were collected, milled, air-dried, pulverized, and subjected to maceration extraction with water as a solvent. The total phenolic, flavonoid contents, and antioxidant activity of the aqueous extract were evaluated using the Folin-Ciocalteu reagent (FCR), aluminum chloride colorimetric assay, ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays, respectively. ResultThe total phenolic content and the total flavonoid content of the plant extract were found to be 540.28 and 40.81 mg/100 g dry weight, respectively. The results from DPPH radical scavenging and FRAP assays revealed that the C. microphyllum extract exhibited strong antioxidant activity. The antibacterial property of the aqueous leaf extract was found to be biologically inactive against the selected bacterial strains. However, its complex with Cobalt(II) demonstrated significant antibacterial activity against all the tested strains. ConclusionThis medicinal plant has been found to contain antioxidant components that can be utilized in the treatment and prevention of infectious diseases after proper standardization. In addition, this work showed the possibility to develop antibiotics from biologically inactive naturally occurring substances by coordination with metal ions.