Quantitative Determination of Myricetin in Rat Plasma by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry and its Absolute Bioavailability

Abstract

Myricetin is a widely distributed bioactive flavonoid with scientific interest attributed to its anti-oxidant, antitumor, and anti-inflammatory properties. A specific and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for identification and quantification of myricetin in rat plasma after oral and intravenous administrations. Kaempferol was used as an internal standard. Followed by β-glucuronidase and sulfatase hydrolysis and liquid-liquid extraction with ethyl acetate, the analytes were separated on an Acquity UPLC BEH C18 column (2.1×50 mm, 1.7 μm) and analyzed in the selected ion recording with a negative electrospray ionization mode. The developed method was validated for selectivity, accuracy, precision, linearity, recovery, stability and matrix effect. The assay was validated over a wide concentration range of 2-4,000 ng/mL. Intra- and inter-day precisions were all less than 13.49% and accuracy ranged from 95.75 to 109.80%. The present method was successfully applied to investigate a pharmacokinetic study of myricetin following intravenous and oral administrations to rats. The absolute bioavailability was found to be 9.62% and 9.74% at 2 oral doses (50 mg/kg and 100 mg/kg, respectively), which indicated myricetin was poorly absorbed after oral administration. To our knowledge, this is the first pharmacokinetic evaluation of myricetin as a single active pharmaceutical ingredient in preclinical studies.

Preparation and evaluation of chitosan-based nanogels/gels for oral delivery of myricetin

“…Blood samples were collected from the oculi chorioideae vein into a heparinized tube at predetermined time points of 0 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h, and 48 h. After centrifugation at 13,000 rpm for 10 min, the plasma layer (100 µL) was immediately transferred to clean tubes and stored at -80°C until analysis. Plasma preparation and Myr quantification were performed as described by Dang et al (2014).…”

“…As a natural antioxidant, Myr has a variety of therapeutic applications including as a cardiovascular protective agent, a hepatoprotective agent, and a potential agent for the treatment of colorectal carcinoma (Dajas et al, 2003;Liu et al, 2007;Maheshwari et al, 2011). However, Myr is essentially insoluble in water (16.60 μg·mL -1 ) and has a slow intrinsic dissolution rate of 11.66 ± 0.82 g·min -1 ·cm -2 and a rapid degradation rate of 0.1860 ± 0.0094 h -1 and 0.3343 ± 0.0547 h -1 at pH 4.5 and 6.8, respectively (Yao et al, 2014a), which probably resulted in its poor oral bioavailability in rats (Dang et al, 2014). Fortunately, the solubility and the dissolution rate of Myr were greatly A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT 5 enhanced by forming a Myr/hydroxypropyl-beta-cyclodextrin (HP-β-CD) inclusion complex, and accordingly the in vitro antioxidant activity and in vivo oral bioavailability in rats were effectively improved.…”

Solid-State and Solution Characterization of Myricetin

“…Quercetin, a flavonoid similar in structure to MYR, has been reported to be fairly insoluble at room temperature (5.5 μg/ml) (8). Likewise, MYR has also been reported to have low aqueous solubility at room temperature (<1.5 μg/ml) (9) while in vivo experimentation indicated a low bioavailability of 9.6% in rats (10).…”

Effects of stabilizing agents on the development of myricetin nanosuspension and its characterization: An in vitro and in vivo evaluation

“…For example, the oral bioavailability of quercetin in humans (Hollman et al, 1996) was only 1%, which was ascribed to, at least in part, its low aqueous solubility of 2.84 AE 0.03 mg/mL (Kakran et al, 2012); silybin is an insoluble flavonoid compound (0.4 mg/mL), and its clinical efficacy is discounted by low absorption in the gastrointestinal tract (23-47%) (Woo et al, 2007;Wu et al, 2007); and the uses of baicalein as a pharmaceutical are limited by its low water solubility (approximately 0.1 mg/mL) and poor oral bioavailability (Hsiu et al, 2002). As for myricetin, its low absolute bioavailability (less than 10% in rats) was also attributed to its poor aqueous solubility (2 mg/mL) (Dang et al, 2014;Yao et al, 2014), which restrained its pharmaceutical development and further clinical application. Therefore, an alternative oral formulation of myricetin with an enhanced solubility and improved bioavailability is highly desired in order to fully realize its therapeutic effects in clinic.…”