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Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma.

Singh G, Pai RS, Pandit V - J Adv Pharm Technol Res (2012)

Bottom Line: This method was linear over the range of 0.010 to 6.4 μg/ml with regression coefficient greater than 0.9998.Resveratrol was found to be stable for a period of 15 days on storage at -20°C.The method was found to be precise, accurate, and specific during the study.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Al-Ameen College of Pharmacy, Bangalore, Karnataka, India.

ABSTRACT
A simple, accurate, precise, sensitive, and reproducible high-performance liquid chromatography method was developed for the determination of Resveratrol (trans-3, 4',5-trihydroxystilbene) in human plasma using liquid-liquid extraction. Caffeine was employed as an internal standard (IS). However, little information is known about its distribution in the organism generally because of the lack of accurate and precise detection methods. The chromatographic separation was achieved on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm) at room temperature in isocratic mode, and the column effluent was monitored by UV detector at 306 nm. The mobile phase used was methanol: phosphate buffer (pH 6.8 adjusted with 0.5% (v/v) orthophosphoric acid solution in Milli-Q water) (63:37%, v/v) at a flow rate of 1.0 ml/min. Nominal retention times of trans-resveratrol and IS were 3.94 and 7.86 minutes, respectively. Limits of detection and Limits of quantification of trans-resveratrol were 0.006 μg/ml and 0.008 μg/ml, respectively. This method was linear over the range of 0.010 to 6.4 μg/ml with regression coefficient greater than 0.9998. The inter- and intra-day precisions in the samples, 0.010, 3.2 and 6.4 μg/ml of trans-resveratrol was in the range 0.63 to 2.12% relative standard deviation (RSD) and 0.46 to 1.02% RSD, respectively. Resveratrol was found to be stable for a period of 15 days on storage at -20°C. The method was found to be precise, accurate, and specific during the study.

No MeSH data available.


Related in: MedlinePlus

HPLC chromatograms of human plasma spiked with trans-resveratrol and caffeine
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Figure 3: HPLC chromatograms of human plasma spiked with trans-resveratrol and caffeine

Mentions: The selectivity was studied by analyzing blank plasma samples. The chromatogram of blank plasma [Figure 2] did not show any interfering compound. A typical chromatogram of a drug-free plasma sample spiked with trans-resveratrol and IS is shown in Figure 3. The retention times of trans-resveratrol and IS were 3.94 and 7.86 minutes, respectively.


Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma.

Singh G, Pai RS, Pandit V - J Adv Pharm Technol Res (2012)

HPLC chromatograms of human plasma spiked with trans-resveratrol and caffeine
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3401675&req=5

Figure 3: HPLC chromatograms of human plasma spiked with trans-resveratrol and caffeine
Mentions: The selectivity was studied by analyzing blank plasma samples. The chromatogram of blank plasma [Figure 2] did not show any interfering compound. A typical chromatogram of a drug-free plasma sample spiked with trans-resveratrol and IS is shown in Figure 3. The retention times of trans-resveratrol and IS were 3.94 and 7.86 minutes, respectively.

Bottom Line: This method was linear over the range of 0.010 to 6.4 μg/ml with regression coefficient greater than 0.9998.Resveratrol was found to be stable for a period of 15 days on storage at -20°C.The method was found to be precise, accurate, and specific during the study.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Al-Ameen College of Pharmacy, Bangalore, Karnataka, India.

ABSTRACT
A simple, accurate, precise, sensitive, and reproducible high-performance liquid chromatography method was developed for the determination of Resveratrol (trans-3, 4',5-trihydroxystilbene) in human plasma using liquid-liquid extraction. Caffeine was employed as an internal standard (IS). However, little information is known about its distribution in the organism generally because of the lack of accurate and precise detection methods. The chromatographic separation was achieved on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm) at room temperature in isocratic mode, and the column effluent was monitored by UV detector at 306 nm. The mobile phase used was methanol: phosphate buffer (pH 6.8 adjusted with 0.5% (v/v) orthophosphoric acid solution in Milli-Q water) (63:37%, v/v) at a flow rate of 1.0 ml/min. Nominal retention times of trans-resveratrol and IS were 3.94 and 7.86 minutes, respectively. Limits of detection and Limits of quantification of trans-resveratrol were 0.006 μg/ml and 0.008 μg/ml, respectively. This method was linear over the range of 0.010 to 6.4 μg/ml with regression coefficient greater than 0.9998. The inter- and intra-day precisions in the samples, 0.010, 3.2 and 6.4 μg/ml of trans-resveratrol was in the range 0.63 to 2.12% relative standard deviation (RSD) and 0.46 to 1.02% RSD, respectively. Resveratrol was found to be stable for a period of 15 days on storage at -20°C. The method was found to be precise, accurate, and specific during the study.

No MeSH data available.


Related in: MedlinePlus