The terrestrial Ludisia discolor, also referred to as the jewel orchid is prized for
the quality of its leaves. L. discolor is known as a medicinal herb and is touted for its heatand
pathogen-resisting qualities. L. discolor is valuable in the production of both flavonoids
and anthocyanins, antioxidants that are exalted in the health industry. Plant cell cultures
have emerged as alternative sources of anthocyanin production. Plant protoplast cultures
are used frequently in transient gene expression studies and in the establishment of callus
and cell suspension cultures. Benefits of plant protoplast system include similarity to cells
found in plant tissues, reproduction under controlled conditions, and prevention of masking
of stress responses to previous handling techniques. A study was conducted to assess the
amenability of the stem and leaves of L. discolor to protoplast isolation. The stem and leaf
segments were weighed, sliced into thin layers, immersed in a digestion medium, washed
and then cultured onto a recovery medium. Results indicated that the production of plant
protoplasts from L. discolor may be viewed as an alternative in the generation of cell
cultures and ultimately in the production of anthocyanins from the cell cultures.
Clitoria ternatea is a herbaceous plant with many health benefits. Extraction is crucial to obtain its bioactive components which contribute to its antioxidant properties. Therefore, this study was conducted to find an optimum extraction condition of C. ternatea flower on total phenolic content (TPC) and antioxidant activity (2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity) as well as to determine its total flavonoid content (TFC) and anthocyanin content based on the optimum extraction condition generated by Response Surface Methodology (RSM)-Design Expert 7.1.5. TPC, TFC and total anthocyanin of C. ternatea were conducted by Folin Ciocalteu (FC), calorimetric assay and pH differential method, respectively. The ranges of selected independent variables were ethanol concentration (30°C-90% v/v), time (60-120 min) and temperature (30°C-70°C). The optimum extraction condition was obtained at 39.62% v/v ethanol concentration, 90 min and 44.24°C. However, these values were slightly adjusted according to the convenience of equipment to operate in which ethanol concentration was adjusted to 37% v/v, time remain at 90 min and temperature at 45°C. The predicted values of TPC and DPPH radical scavenging activity were 41.60 mg GAE/g dry samples and 68.12% inhibition and were experimentally verified to be 41.17 ± 0.5 mg GAE/g dry samples and 63.53 ± 0.95% inhibition of TPC and DPPH radical scavenging activity respectively. This result has showed RSM can optimise TPC and radical scavenging activity of C. ternatea. Upon the optimum condition, the TFC determined was 187.05 ± 3.18 mg quercetin/g dried sample which was higher than TPC and the total anthocyanin content was 28.60 ± 0.04 mg/L. Hence, the extractable phenolic, flavonoid and anthocyanin compounds indicated that C. ternatea is a good source of natural antioxidant.
Anthocyanins not just have various benefits in food industry but also have been used as natural colourants in cosmetic, coating products and as potential natural photosensitizers in solar cell. Thus, the main purpose of this study was to obtain information on the maximum yield of anthocyanin that can be recovered from Melastoma malabathricum fruit. Factors such as extraction temperature, extraction time, and solid to liquid ratio were identified to be significantly affecting anthocyanin extraction efficiency. By using three-level three-factor Box-Behnken design, the optimized conditions for anthocyanin extraction by acidified methanol (R (2) = 0.972) were temperature of 60°C, time of 86.82 min, and 0.5 : 35 (g/mL) solid to liquid ratio while the optimum extraction conditions by acidified ethanol (R (2) = 0.954) were temperature of 60°C, time of 120 min, and 0.5 : 23.06 (g/mL) solid to liquid ratio. The crude anthocyanin extract was further purified by using Amberlite XAD-7 and Sephadex LH-20 column chromatography. Identification of anthocyanins revealed the presence of cyanidin dihexoside, cyanidin hexoside, and delphinidin hexoside as the main anthocyanins in M. malabathricum fruit.
A mimicked biosynthetic pathway of catechin metabolite genes from C. sinensis, consisting of flavanone 3 hydroxylase (F3H), dihydroflavonol reductase (DFR), and leucoanthocyanidin reductase (LCR), was designed and arranged in two sets of constructs: (a) single promoter in front of F3H and ribosome-binding sequences both in front of DFR and LCR; (b) three different promoters with each in the front of the three genes and ribosome-binding sequences at appropriate positions. Recombinant E. coli BL (DE3) harbouring the constructs were cultivated for 65 h at 26 °C in M9 medium consisting of 40 g/L glucose, 1 mM IPTG, and 3 mM eriodictyol. Compounds produced were extracted in ethyl acetate in alkaline conditions after 1 h at room temperature and identified by HPLC. Two of the four major catechins, namely, (-)-epicatechin (0.01) and (-)-epicatechin gallate (0.36 mg/L), and two other types ((+)-catechin hydrate (0.13 mg/L) and (-)-catechin gallate (0.04 mg/L)) were successfully produced.
The natural dyes anthocyanin and chlorophyll were extracted from Musa acuminata bracts and Alternanthera dentata leaves, respectively. The dyes were then applied as sensitizers in TiO2-based dye-sensitized solar cells (DSSCs). The ethanol extracts of the dyes had maximum absorbance. High dye yields were obtained under extraction temperatures of 70 to 80°C, and the optimal extraction temperature was approximately 80°C. Moreover, dye concentration sharply decreased under extraction temperatures that exceeded 80°C. High dye concentrations were obtained using acidic extraction solutions, particularly those with a pH value of 4. The DSSC fabricated with anthocyanin from M. acuminata bracts had a conversion efficiency of 0.31%, short-circuit current (Isc) of 0.9mA/cm2, open-circuit voltage (Voc) of 0.58V, and fill factor (FF) of 62.22%. The DSSC sensitized with chlorophyll from A. dentata leaves had a conversion efficiency of 0.13%, Isc of 0.4mA/cm-2,Voc of 0.54V, and FF of 67.5%. The DSSC sensitized with anthocyanin from M. acuminata bracts had a maximum incident photon-to-current conversion efficiency of 42%, which was higher than that of the DSSC sensitized with chlorophyll from A. dentata leaves (23%). Anthocyanin from M. acuminata bracts exhibited the best photosensitization effects.
A developed colorimetric pH sensor film based on edible materials for real-time monitoring of food freshness is described. The mixed natural dyes from edible plants Clitoria sp and Brassica sp were extracted and incorporated into ι-carrageenan film as a colorimetric pH sensor film for monitoring food spoilage and its freshness. The color changes of the developed colorimetric sensor film were measured with chromametry and UV-vis spectroscopy, respectively. Experimental results show that colorimetric pH sensor film demonstrated statistically significant differences (p < 0.05) between CIE-L*a*b* coordinates color system indicated that the developed colorimetric sensor film was able to give a gradual change in color over a wide pH range. The color of the colorimetric sensor film also changes discretely and linearly with factors that contribute to food spoilage using shrimp and durian samples. Moreover, the developed colorimetric pH sensor film has the potential to be used as a safe, non-destructive testing and also a flexibly visual method for direct assessment of food freshness indicator during storage.
Clitoria ternatea flower is a traditional medicinal herb that has been used as a natural food colourant. As there are limited studies on investigating the bioactivities of the anthocyanin-rich fraction of Clitoria ternatea flower, this study aimed to determine an efficient column chromatography method to obtain the anthocyanin-rich fraction from this flower and characterise its composition, antioxidant, antibacterial, and cytotoxic activities. Amberlite XAD-16 column chromatography was more efficient in enriching the total anthocyanin content (TAC) of the fraction with the highest TAC to total phenolic content (TPC) ratio of 1:6 than that using C18-OPN. A total of 11 ternatin anthocyanins were characterised in the anthocyanin-rich fraction by LC-MS analysis. The antioxidant activity of the anthocyanin-rich fraction was more potent in the chemical-based assay with an IC50 value of 0.86 ± 0.07 mg/mL using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay than cellular antioxidant assay using RAW 264.7 macrophages. In vitro cytotoxicity assay using human embryonic kidney HEK-293 cell line showed the anthocyanin-rich fraction to be more toxic than the crude extracts. The anthocyanin-rich fraction had more potent antibacterial activity than the crude extracts against Bacillus cereus, Bacillus subtilis, and Escherichia coli. The anthocyanin-rich fraction of C. ternatea has the potential to be used and developed as a functional food ingredient or nutraceutical agent.
This study aimed to determine the total phenolics and antioxidant capacity of defatted dabai parts based on liquid extraction and optimized using response surface methodology (RSM). A two-level factorial design was applied to determine the effect of two independent variables (extraction time: X1 and % methanol: X2) on three response variables (total phenolic content: Y1, total flavonoid/anthocyanin content: Y2 and Trolox equivalent antioxidant capacity: Y3). The optimum conditions for extraction time and percent methanol were 36 min or 1 min and 62.25% or 53% for the defatted dabai pulp or peel, respectively. The RSM optimized extraction was compared with sonication-assisted extraction. Optimization results showed that defatted dabai parts had high total phenolic content and antioxidant capacity. Sonication-assisted extraction utilized the optimized extraction conditions had further increased the total phenolic content and antioxidant capacity of defatted dabai peel, but not in the pulp. Therefore, optimization of different extraction methods for the defatted fruit parts is recommended for future studies.
UKMR-1, a local variant of mutant Roselle strain (Hibiscus sabdariffa) is enriched with free radical scavenging polyphenols
such as anthocyanin, vitamin C and hydroxycitric acid. However, pharmacological actions of UKMR-1 are not fully known.
This study was conducted to determine whether supplementation of aqueous UKMR-1 calyx extract was able to protect
against nicotine-induced cardiac injury in rats. In this experimental study, healthy male albino rats were randomly
allotted into three groups (n=7 per group): control, nicotine and UKMR-1+Nicotine groups. Nicotine (0.6 mg/kg, i.p.)
was administered to both nicotine and UKMR-1+Nicotine groups for 28 consecutive days. UKMR-1+Nicotine group also
received 100 mg/kg UKMR-1 extract orally via gavage 30 min prior to nicotine injection, daily. UKMR-1+Nicotine group
had significantly (p<0.05) higher lactate dehydrogenase (LDH) activity, as well as lower malondialdehyde content in
heart tissue homogenate than nicotine group, suggesting its cardio protective activity by inhibition of lipid peroxidation.
UKMR-1 also lowered (p<0.05) the blood pressure in nicotine-administered rats. In addition, UKMR-1 significantly (p<0.05)
restored activities of cytosolic superoxide dismutase, glutathione peroxidase and glutathione-S-transferase as well as
redox balance ratio (GSH:GSSG). In conclusion, UKMR-1 was a
Melastoma malabathricum, belongs to the Melastomaceae family, is an important medicinal plant widely distributed from Madagascar to Australia, that is used in traditional remedies for the treatment of various ailments. Besides its medicinal properties, it has been identified as a potential source of anthocyanin production. The present study was carried out to investigate the effect of sucrose and methyl jasmonate and feeding time on cell biomass yield and anthocyanin production in cell suspension culture of M. malabathricum. Addition of different concentrations of sucrose into the cell culture of M. malabathricum influenced cell biomass and pigment accumulation. The addition of methyl jasmonate was found to have no effect on cell biomass but the presence of higher amount (12.5-50 mg/L) had caused a reduction in anthocyanin production and accumulation. MS medium supplemented with 30 g/L sucrose and 3.5 mg/L of MeJA added on cero day and 3rd day produced high fresh cell mass at the end of nine days of culture but did not support the production of anthocyanins. However, cells cultured in the medium supplemented with 45 g/L sucrose without MeJA showed the highest pigment content (0.69 +/- 0.22 CV/g-FCM). The cells cultured in MS medium supplemented with 30 g/L sucrose with 3.5 mg/L MeJA added on the 3rd and 6th day of culture, showed the lowest pigment content (0.37-0.40 CV/g-FCM). This study indicated that MeJA was not necessary but sucrose was needed for the enhancement of cell growth and anthocyanin production in M. malabathricum cell cultures.
A simple optical pH sensor based on immobilization, Dioscorea alata L. anthocyanin methanol extract, onto a pectin-chitosan polyelectrolyte complex (pectin-chitosan PEC), has been successfully fabricated. The optical pH sensor was manufactured as a membrane made of pectin-chitosan PEC and the extracted anthocyanin. This sensor has the highest sensitivity of anthocyanin content at 0.025 mg/L in phosphate buffer and 0.0375 mg/L in citrate buffer. It also has good reproducibility with a relative standard deviation (%RSD) of 7.7%, and gives a stable response at time values greater than 5 min from exposure in a buffer solution, and the sensor can be utilized within five days from its synthesis. This optical pH sensor has been employed to determine saliva pH of people of different ages and showed no significant difference when compared to a potentiometric method.
Canarium odontophyllum, also known as CO, is a highly nutritious fruit. Defatted parts of CO fruit are potent sources of nutraceutical. This study aimed to determine oxidative stress and lipid peroxidation effects of defatted CO pericarp and peel extracts using in vitro bioassays. Cell cytotoxic effect of the CO pericarp and peel extracts were also evaluated using HUVEC and Chang liver cell lines. The crude extracts of defatted CO peel and pericarp showed cytoprotective effects in t-BHP and 40% methanol-induced cell death. The crude extracts also showed no toxic effect to Chang liver cell line. Using CD36 ELISA, NAD(+) and LDL inhibition assays, inhibition of oxidative stress were found higher in the crude extract of defatted CO peel compared to the pericarp extract. Hemoglobin and LDL oxidation assays revealed both crude extracts had significantly reduced lipid peroxidation as compared to control. TBARS values among defatted CO pericarp, peel, and cyanidin-3-glucoside showed no significant differences for hemoglobin and LDL oxidation assays. The protective effects of defatted CO parts, especially its peel is related to the presence of high anthocyanin that potentially offers as a pharmaceutical ingredient for cardioprotection.
Anoectochilus sp. and Ludisia discolor are known as Jewel orchids. Both species are terrestrial wild orchids that grow in shaded areas of forests. The Jewel orchids are renowned for the beauty of their leaves, which are dark-green laced with silvery or golden veins. The orchids are used as a cure in various parts of Asia. Overharvesting and anthropogenic disturbances threaten the existence of the Jewel orchids in the wild, necessitating human intervention in their survival. An understanding of the structure and adaptations of a plant may assist in its survival when propagated outside of its habitat. In this study, ex vitro leaves of Anoectochilus sp. and L. discolor were subjected to freehand sectioning, and then inspected through brightfield and fluorescence microscopy. The study indicated that all parts of both plants presented typical monocotyledonous characteristics except the leaves. The leaves displayed dorsiventrality with distinct palisade and spongy mesophyll layers. The spongy mesophyll layer contained cells which fluoresced a bright red when exposed to ultraviolet, blue, and green light wavelengths, hinting at the presence of anthocyanins for photoprotection. Cyanidin was detected in the leaves of L. discolor, as enumerated through high performance liquid chromatography (HPLC). The observations indicated that Anoectochilus sp. and L. discolor are well-adapted to live under shaded conditions with minimal exposure to light.
Passiflora quadrangularis L. belongs to the family Passifloraceae which bears larger fruit with edible juicy mesocarp and pulp known as a good source of phytochemicals. Cultivation and plant management practices are known to influence the phytochemical compositions of agricultural produce. This study aimed to examine the influence of the cultivation practices on the antioxidant activities and secondary metabolites of the organically and conventionally grown P. quadrangularis. Findings revealed organically treated P. quadrangularis plants showed enhancement in their antioxidant properties and secondary metabolites profiles. Among the plant parts, leaves of P. quadrangularis grown organically possessed higher antioxidant activities compared to the conventional in all assays evaluated. The antioxidant activities in the edible parts of the P. quadrangularis fruit have also been enhanced through organic cultivation with significantly higher total phenolic content and DPPH in mesocarp, and the pulp showed higher total flavonoid content, DPPH and FRAP. This observation is supported by a higher level of vitamins and secondary metabolites in the samples. The secondary metabolites profile showed mesocarps were phenolic rich, the pulps were flavonoids rich while leaves showed good composition of phenolics, flavonoids and terpenoids with outstanding antioxidant activities. The common secondary metabolites for organically produced P. quadrangularis in different plant parts include 2-isopropyl-3-methoxycinnamic acid (mesocarp and pulp), myricetin isomers (pulp and leaves), and malvidin-3-O-arabinoside isomers (pulp and leaves). This study confirmed that organic cultivated P. quadrangularis possessed higher antioxidant activities contributed by its vitamins and secondary metabolites.
Quinoa (Chenopodium quinoa Willd.) has gained significant popularity among agricultural scientists and farmers throughout the world due to its high nutritive value. It is cultivated under a range of soil and climatic conditions; however, late sowing adversely affects its productivity and yield due to shorter growth period. Inorganic and organic phyto-stimulants are promising for improving growth, development, and yield of field crops under stressful environments. Field experiments were conducted during crop cultivation seasons of 2016-17 and 2017-18, to explore the role of inorganic (hydrogen peroxide and ascorbic acid) and organic [moringa leaf extract (MLE) and sorghum water extract (sorgaab)] phyto-stimulants in improving growth and productivity of quinoa (cultivar UAF-Q7). Hydrogen peroxide at 100 μM, ascorbic acid at 500 μM, MLE at 3% and sorgaab at 3% were exogenously applied at anthesis stage of quinoa cultivated under normal (November 21st and 19th during 2016 and 2017) and late-sown (December 26th and 25th during 2016 and 2017) conditions. Application of inorganic and organic phyto-stimulants significantly improved biochemical, physiological, growth and yield attributes of quinoa under late sown conditions. The highest improvement in these traits was recorded for MLE. Application of MLE resulted in higher chlorophyll a and b contents, stomatal conductance, and sub-stomatal concentration of CO2 under normal and late-sowing. The highest improvement in soluble phenolics, anthocyanins, free amino acids and proline, and mineral elements in roots, shoot and grains were observed for MLE application. Growth attributes, including plant height, plant fresh weight and panicle length were significantly improved with MLE application as compared to the rest of the treatments. The highest 1000-grain weight and grain yield per plant were noted for MLE application under normal and late-sowing. These findings depict that MLE has extensive crop growth promoting potential through improving physiological and biochemical activities. Hence, MLE can be applied to improve growth and productivity of quinoa under normal and late-sown conditions.
The growing demand for high value aromatic herb Polygonum minus-based products have increased in recent years, for its antioxidant, anticancer, antimicrobial, and anti-inflammatory potentials. Although few reports have indicated the chemical profiles and antioxidative effects of Polygonum minus, no study has been conducted to assess the benefits of micro-environmental manipulation (different shading levels) on the growth, leaf gas exchange and secondary metabolites in Polygonum minus. Therefore, two shading levels (50%:T2 and 70%:T3) and one absolute control (0%:T1) were studied under eight weeks and 16 weeks of exposures on Polygonum minus after two weeks. It was found that P. minus under T2 obtained the highest photosynthesis rate (14.892 µmol CO2 m-2 s-1), followed by T3 = T1. The increase in photosynthesis rate was contributed by the enhancement of the leaf pigments content (chlorophyll a and chlorophyll b). This was shown by the positive significant correlations observed between photosynthesis rate with chlorophyll a (r2 = 0.536; p ≤ 0.05) and chlorophyll b (r2 = 0.540; p ≤ 0.05). As the shading levels and time interval increased, the production of total anthocyanin content (TAC) and antioxidant properties of Ferric Reducing Antioxidant Power (FRAP) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) also increased. The total phenolic content (TPC) and total flavonoid content (TFC) were also significantly enhanced under T2 and T3. The current study suggested that P.minus induce the production of more leaf pigments and secondary metabolites as their special adaptation mechanism under low light condition. Although the biomass was affected under low light, the purpose of conducting the study to boost the bioactive properties in Polygonum minus has been fulfilled by 50% shading under 16 weeks' exposure.
The present study represents the first report of the effect of hydrogen peroxide (H(2)O(2)) on the growth, development and quality of the wax apple fruit, a widely cultivated fruit tree in South East Asia. The wax apple trees were spray treated with 0, 5, 20 and 50 mM H(2)O(2) under field conditions. Photosynthetic rates, stomatal conductance, transpiration, chlorophyll and dry matter content of the leaves and total soluble solids and total sugar content of the fruits of wax apple (Syzygium samarangense, var. jambu madu) were significantly increased after treatment with 5 mM H(2)O(2). The application of 20 mM H(2)O(2) significantly reduced bud drop and enhanced fruit growth, resulting in larger fruit size, increased fruit set, fruit number, fruit biomass and yield compared to the control. In addition, the endogenous level of H(2)O(2) in wax apple leaves increased significantly with H(2)O(2) treatments. With regard to fruit quality, 20 mM H(2)O(2) treatment increased the K(+), anthocyanin and carotene contents of the fruits by 65%, 67%, and 41%, respectively. In addition, higher flavonoid, phenol and soluble protein content, sucrose phosphate synthase (SPS), phenylalanine ammonia lyase (PAL) and antioxidant activities were recorded in the treated fruits. There was a positive correlation between peel colour (hue) and TSS, between net photosynthesis and SPS activity and between phenol and flavonoid content with antioxidant activity in H(2)O(2)-treated fruits. It is concluded that spraying with 5 and 20 mM H(2)O(2) once a week produced better fruit growth, maximising the yield and quality of wax apple fruits under field conditions.
Anthocyanins and volatile phenylpropenes (isoeugenol and eugenol) in petunia (Petunia hybrida) flowers have the precursor 4-coumaryl coenzyme A (CoA) in common. These phenolics are produced at different stages during flower development. Anthocyanins are synthesized during early stages of flower development and sequestered in vacuoles during the lifespan of the flowers. The production of isoeugenol and eugenol starts when flowers open and peaks after anthesis. To elucidate additional biochemical steps toward (iso)eugenol production, we cloned and characterized a caffeoyl-coenzyme A O-methyltransferase (PhCCoAOMT1) from the petals of the fragrant petunia 'Mitchell'. Recombinant PhCCoAOMT1 indeed catalyzed the methylation of caffeoyl-CoA to produce feruloyl CoA. Silencing of PhCCoAOMT1 resulted in a reduction of eugenol production but not of isoeugenol. Unexpectedly, the transgenic plants had purple-colored leaves and pink flowers, despite the fact that cv Mitchell lacks the functional R2R3-MYB master regulator ANTHOCYANIN2 and has normally white flowers. Our results indicate that down-regulation of PhCCoAOMT1 activated the anthocyanin pathway through the R2R3-MYBs PURPLE HAZE (PHZ) and DEEP PURPLE, with predominantly petunidin accumulating. Feeding cv Mitchell flowers with caffeic acid induced PHZ expression, suggesting that the metabolic perturbation of the phenylpropanoid pathway underlies the activation of the anthocyanin pathway. Our results demonstrate a role for PhCCoAOMT1 in phenylpropene production and reveal a link between PhCCoAOMT1 and anthocyanin production.
Objectives: Polyphenols, particularly anthocyanins, have received attention in improving health issues during old age, including decline in cognitive function and other health parameters. We aimed to determine the effects of polyphenols-rich tropical fruit TP 3-in-1™ juice towards improving cognitive function, oxidative stress and metabolomics profiles among middle-aged women.Methods: This clinical trial involved 31 subjects with signs of poor cognitive function, as assessed using the Rey Auditory Verbal Learning Test (RAVLT). They were randomized to receive either TP 3-in-1™ juice (n = 16) or placebo (n = 15). Study subjects consumed 500 ml of beverages for three times per day, three days per week, for a period of ten weeks. Juice supplementation provided 9135 mg GAE of total phenolic content and 194.1 mg cyanidin-3-glucoside of total anthocyanin monomer.Results: There was a significant interaction effects on RAVLT immediate recall (p
The ultrasonic extraction (UE) method of anthocyanin from Clitoria ternatea flowers using response surface methodology (RSM) was performed in this study. By using RSM, the objective is to optimise the extraction yield of anthocyanin from C. ternatea which is influenced by various factors, including the extraction temperature, time, ratio of solvent to solid and ultrasonic power. The empirical model was investigated by performing first-level optimisation in a two-level factorial design with Design Expert 7 software. In comparison with the conventional solvent extraction, UE showed a 246.48% better extraction yield and produced an anthocyanin extract with a radical scavenging activity of 68.48% at the optimised factors of 50°C, 150 min, 15 mL/g and 240 W.