Optimized DPPH assay in a detergent-based buffer system for measuring antioxidant activity of proteins

Nicklisch, Sascha; Waite, J. Herbert

doi:10.1016/j.mex.2014.10.004

Cited by 55 publications

(32 citation statements)

References 20 publications

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“…Using the DPPH assay, we demonstrated that antioxidant activity solubilized from plaques and the degree of antioxidant activity indirectly correlated to plaque age (significant ( p , 0.001) decrease in antioxidant activity between days 1-4 and days [17][18][19][20]. Antioxidant activity (normalized % of antioxidant activity [(sample DPPH activity 2 negative control)/full DPPH reduction] Â 100) was maintained at 60-70% over days 1-12, and then stabilized at approximately 30% over days [13][14][15][16][17][18][19][20] (no significant ( p .…”

Section: Antioxidant Activity In Plaque Extracts and In Situmentioning

confidence: 97%

“…Antioxidant activity (normalized % of antioxidant activity [(sample DPPH activity 2 negative control)/full DPPH reduction] Â 100) was maintained at 60-70% over days 1-12, and then stabilized at approximately 30% over days [13][14][15][16][17][18][19][20] (no significant ( p . 0.87) change between the days 13-16 and days 17-20, n ¼ 13-20, figure 2c).…”

Section: Antioxidant Activity In Plaque Extracts and In Situmentioning

confidence: 99%

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The staying power of adhesion-associated antioxidant activity inMytilus californianus

Miller

Spahn

Waite

2015

J. R. Soc. Interface.

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The California mussel, Mytilus californianus, adheres in the highly oxidizing intertidal zone with a fibrous holdfast called the byssus using 3, 4-dihydroxyphenyl-L-alanine (DOPA)-containing adhesive proteins. DOPA is susceptible to oxidation in seawater and, upon oxidation, loses adhesion. Successful mussel adhesion thus depends critically on controlling oxidation and reduction. To explore how mussels regulate redox during their functional adhesive lifetime, we tracked extractable protein concentration, DOPA content and antioxidant activity in byssal plaques over time. In seawater, DOPA content and antioxidant activity in the byssus persisted much longer than expected-50% of extractable DOPA and 30% of extractable antioxidant activity remained after 20 days. Antioxidant activity was located at the plaque-substrate interface, demonstrating that antioxidant activity keeps DOPA reduced for durable and dynamic adhesion. We also correlated antioxidant activity to cysteine and DOPA side chains of mussel foot proteins (mfps), suggesting that mussels use both cysteine and DOPA redox reservoirs for controlling interfacial chemistry. These data are discussed in the context of the biomaterial structure and properties of the marine mussel byssus.

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Section: Antioxidant Activity In Plaque Extracts and In Situmentioning

confidence: 97%

Section: Antioxidant Activity In Plaque Extracts and In Situmentioning

confidence: 99%

The staying power of adhesion-associated antioxidant activity inMytilus californianus

Miller

Spahn

Waite

2015

J. R. Soc. Interface.

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“…Briefly, 150 µL of formulated flavonosomes or PC or QKA or QKA + PC was mixed with 50 µL of 0.1 mM DPPH solution (methanol with 0.3% Triton X-100 v/v). 30 The flavonoids/PC content in carrier, free flavonoids mix, and physical mixture were adjusted to the same concentration as the formulated flavonosomes. Then the solution was incubated at 37°C in the dark for regulated time points at …”

Section: In Vitro Antioxidant Dpph Kineticsmentioning

confidence: 99%

“…Free DPPH radical at a concentration of 0.1 mM dissolved in methanolic solution with 0.3% Triton X-100 v/v was stable for up to 16 hours in the reaction medium. 30 Thus, it allows the compounds to be evaluated in that time range. In this study, we have evaluated the scavenging activity for up to 4 hours at various time points by using individual experimental set for each time point.…”

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confidence: 99%

Optimization, formulation, and characterization of multiflavonoids-loaded flavanosome by bulk or sequential technique

Karthivashan¹,

Kura²,

Abas³

et al. 2016

IJN

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This study involves adaptation of bulk or sequential technique to load multiple flavonoids in a single phytosome, which can be termed as “flavonosome”. Three widely established and therapeutically valuable flavonoids, such as quercetin (Q), kaempferol (K), and apigenin (A), were quantified in the ethyl acetate fraction of Moringa oleifera leaves extract and were commercially obtained and incorporated in a single flavonosome (QKA–phosphatidylcholine) through four different methods of synthesis – bulk (M1) and serialized (M2) co-sonication and bulk (M3) and sequential (M4) co-loading. The study also established an optimal formulation method based on screening the synthesized flavonosomes with respect to their size, charge, polydispersity index, morphology, drug–carrier interaction, antioxidant potential through in vitro 1,1-diphenyl-2-picrylhydrazyl kinetics, and cytotoxicity evaluation against human hepatoma cell line (HepaRG). Furthermore, entrapment and loading efficiency of flavonoids in the optimal flavonosome have been identified. Among the four synthesis methods, sequential loading technique has been optimized as the best method for the synthesis of QKA–phosphatidylcholine flavonosome, which revealed an average diameter of 375.93±33.61 nm, with a zeta potential of −39.07±3.55 mV, and the entrapment efficiency was >98% for all the flavonoids, whereas the drug-loading capacity of Q, K, and A was 31.63%±0.17%, 34.51%±2.07%, and 31.79%±0.01%, respectively. The in vitro 1,1-diphenyl-2-picrylhydrazyl kinetics of the flavonoids indirectly depicts the release kinetic behavior of the flavonoids from the carrier. The QKA-loaded flavonosome had no indication of toxicity toward human hepatoma cell line as shown by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide result, wherein even at the higher concentration of 200 µg/mL, the flavonosomes exert >85% of cell viability. These results suggest that sequential loading technique may be a promising nanodrug delivery system for loading multiflavonoids in a single entity with sustained activity as an antioxidant, hepatoprotective, and hepatosupplement candidate.

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“…The DPPH (1, 1-diphenyl-2-picrylhydrazyl) and BSLT (brine shrimp lethality test) are general bioassays for pharmacological activities of active phytochemical components of medicinal plants. The DPPH test is a well established assay for the In vitro determination of antioxidant activity in medicinal plant extracts 7 . BSLT is a general bioassay that appears capable of detecting a broad spectrum of bioactivity present in crude extracts of medicinal plants 8 .…”

Section: Introductionmentioning

confidence: 99%

Phytochemical Screening, Antioxidant and Cytotoxic Activities in Extracts of Different Rhizome Parts From Curcuma Aeruginosa Roxb.

Nurcholis¹,

Khumaida²,

Syukur³

et al. 2015

Int. J. Res. Ayurveda Pharm.

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Crude aqueous, 70% ethanol and 96% ethanol extracts from different part of rhizome of Curcuma aeruginosa RoxB., namely primary rhizome (PR), secondary rhizome (SR) and tertiary rhizome (TR) were screened for phytochemical compounds using qualitative methods, antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and cytotoxicity using brine shrimp lethality test (BSLT). The crude 70% ethanol extract of PR (IC50, 437.07 µg/ ml) had stronger DPPH scavenging activity compared to the other crude extracts of different part of rhizomes of C. aeruginosa. None of the extracts from different rhizome parts of C. aeruginosa were found to be more active than standard (ascorbic acid, IC50, 32.91 µg/ ml) for DPPH scavenging activity. In the BSLT, we found than the crude 96% ethanol and 70% ethanol extracts of PR exhibited most activity, which had LC50 values of 90.40 and 265.17 µg/ml, respectively. Phytochemical screening revealed the presence of tannins and triterpenoids in all extracts, and saponins in the 70% ethanol and pure aqueous extracts, but alkaloids, flavonoids, and steroids were absent in all the extracts of different part of rhizome of C. aeruginosa.

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Optimized DPPH assay in a detergent-based buffer system for measuring antioxidant activity of proteins

Abstract: Graphical abstract

Cited by 55 publications

References 20 publications

The staying power of adhesion-associated antioxidant activity inMytilus californianus

The staying power of adhesion-associated antioxidant activity inMytilus californianus

Optimization, formulation, and characterization of multiflavonoids-loaded flavanosome by bulk or sequential technique

Phytochemical Screening, Antioxidant and Cytotoxic Activities in Extracts of Different Rhizome Parts From Curcuma Aeruginosa Roxb.

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