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Resveratrol-loaded polymeric nanoparticles: validation of an HPLC-PDA method to determine the drug entrapment and evaluation of its antioxidant activity.

da Rocha Lindner G, Khalil NM, Mainardes RM - ScientificWorldJournal (2013)

Bottom Line: The mean diameter of the nanoparticles varied between 180 and 220 nm, and the encapsulation efficiency of RVT ranged from 60% to 88%.The profile obtained from the PLA nanoparticles containing RVT demonstrated that after 24 h, there was almost no increase in antioxidant activity, which was lower than that of the free RVT and RVT-loaded PLA-PEG nanoparticles.For PLA-PEG nanoparticles, the radical-scavenging activity of RVT was shown to increase with time, and after 48 h, it was similar to that observed with free RVT.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Laboratory of Pharmaceutical Nanotechnology, Universidade Estadual do Centro-Oeste/UNICENTRO, Rua Simeão Camargo Varela de Sá 03, 85040-080 Guarapuava, PR, Brazil.

ABSTRACT
Poly(lactic acid) (PLA) and PLA-poly(ethylene glycol) (PLA-PEG) nanoparticles containing resveratrol (RVT) were developed, and their antioxidant activity was evaluated. An analytical method using high performance liquid chromatography (HPLC)/photodiode array (PDA) detection was also developed and validated for RVT determination in nanoparticles. The mobile phase consisted of methanol : water (51 : 49, v/v) flowed at 0.9 mL/min, and the PDA detector was set at wavelength of 306 nm. The mean diameter of the nanoparticles varied between 180 and 220 nm, and the encapsulation efficiency of RVT ranged from 60% to 88%. The nanoparticles containing RVT were evaluated for their ability to scavenge the radical (2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) (ABTS•⁺). The profile obtained from the PLA nanoparticles containing RVT demonstrated that after 24 h, there was almost no increase in antioxidant activity, which was lower than that of the free RVT and RVT-loaded PLA-PEG nanoparticles. For PLA-PEG nanoparticles, the radical-scavenging activity of RVT was shown to increase with time, and after 48 h, it was similar to that observed with free RVT.

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Representative HPLC chromatograms of RVT standard (40 μg/mL) in methanol : water. Mobile phase, methanol : water (51 : 49, v/v); flow rate, 0.9 mL/min; PDA detection wavelength, 306 nm; column temperature, 25°C; injection volume, 20 μL.
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fig1: Representative HPLC chromatograms of RVT standard (40 μg/mL) in methanol : water. Mobile phase, methanol : water (51 : 49, v/v); flow rate, 0.9 mL/min; PDA detection wavelength, 306 nm; column temperature, 25°C; injection volume, 20 μL.

Mentions: Initial runs were performed using a mobile phase mixture of methanol and acetonitrile based on existing methods for RVT quantification in plasma [24]. Various ratios in the isocratic mode were tested, some of which led to the presence of more than one peak (methanol : acetonitrile 3 : 1 and 1.5 : 1). Consequently, water and acetic acid were introduced as a new mobile phase mixture [25, 26]. A few symmetric peaks were achieved with methanol, acetonitrile, water, and acetic acid; however, neither method demonstrated repeatability or accuracy. When acetonitrile and acetic acid were removed and only methanol and water were used, lower tailing and more symmetrical peaks were observed. The best peak with respect to width and symmetry was observed with a mobile phase of water and methanol in the ratio of 49 : 51 (v/v) and a flow rate of 0.9 mL/min. The peak was detected at 6.4 min (Figure 1).


Resveratrol-loaded polymeric nanoparticles: validation of an HPLC-PDA method to determine the drug entrapment and evaluation of its antioxidant activity.

da Rocha Lindner G, Khalil NM, Mainardes RM - ScientificWorldJournal (2013)

Representative HPLC chromatograms of RVT standard (40 μg/mL) in methanol : water. Mobile phase, methanol : water (51 : 49, v/v); flow rate, 0.9 mL/min; PDA detection wavelength, 306 nm; column temperature, 25°C; injection volume, 20 μL.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824335&req=5

fig1: Representative HPLC chromatograms of RVT standard (40 μg/mL) in methanol : water. Mobile phase, methanol : water (51 : 49, v/v); flow rate, 0.9 mL/min; PDA detection wavelength, 306 nm; column temperature, 25°C; injection volume, 20 μL.
Mentions: Initial runs were performed using a mobile phase mixture of methanol and acetonitrile based on existing methods for RVT quantification in plasma [24]. Various ratios in the isocratic mode were tested, some of which led to the presence of more than one peak (methanol : acetonitrile 3 : 1 and 1.5 : 1). Consequently, water and acetic acid were introduced as a new mobile phase mixture [25, 26]. A few symmetric peaks were achieved with methanol, acetonitrile, water, and acetic acid; however, neither method demonstrated repeatability or accuracy. When acetonitrile and acetic acid were removed and only methanol and water were used, lower tailing and more symmetrical peaks were observed. The best peak with respect to width and symmetry was observed with a mobile phase of water and methanol in the ratio of 49 : 51 (v/v) and a flow rate of 0.9 mL/min. The peak was detected at 6.4 min (Figure 1).

Bottom Line: The mean diameter of the nanoparticles varied between 180 and 220 nm, and the encapsulation efficiency of RVT ranged from 60% to 88%.The profile obtained from the PLA nanoparticles containing RVT demonstrated that after 24 h, there was almost no increase in antioxidant activity, which was lower than that of the free RVT and RVT-loaded PLA-PEG nanoparticles.For PLA-PEG nanoparticles, the radical-scavenging activity of RVT was shown to increase with time, and after 48 h, it was similar to that observed with free RVT.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Laboratory of Pharmaceutical Nanotechnology, Universidade Estadual do Centro-Oeste/UNICENTRO, Rua Simeão Camargo Varela de Sá 03, 85040-080 Guarapuava, PR, Brazil.

ABSTRACT
Poly(lactic acid) (PLA) and PLA-poly(ethylene glycol) (PLA-PEG) nanoparticles containing resveratrol (RVT) were developed, and their antioxidant activity was evaluated. An analytical method using high performance liquid chromatography (HPLC)/photodiode array (PDA) detection was also developed and validated for RVT determination in nanoparticles. The mobile phase consisted of methanol : water (51 : 49, v/v) flowed at 0.9 mL/min, and the PDA detector was set at wavelength of 306 nm. The mean diameter of the nanoparticles varied between 180 and 220 nm, and the encapsulation efficiency of RVT ranged from 60% to 88%. The nanoparticles containing RVT were evaluated for their ability to scavenge the radical (2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) (ABTS•⁺). The profile obtained from the PLA nanoparticles containing RVT demonstrated that after 24 h, there was almost no increase in antioxidant activity, which was lower than that of the free RVT and RVT-loaded PLA-PEG nanoparticles. For PLA-PEG nanoparticles, the radical-scavenging activity of RVT was shown to increase with time, and after 48 h, it was similar to that observed with free RVT.

Show MeSH