Pharmaceutics & Pharmacokinetics Division, CSIR-Central Drug Research Institute (CSIR-CDRI), Sitapur Road, Lucknow 226031, India.
Academy of Scientific and Innovative Research, Ghaziabad 201002, India.
Molecules. 2023 Jun 8;28(12):4656. doi: 10.3390/molecules28124656.
is a nutrient-rich plant with a history of use in traditional medicine. It boasts a diverse range of polyphenols, including quercetin, resveratrol, β-sitosterol, myricetin, and other compounds. We developed and validated a sensitive LC-MS/MS method to quantify quercetin and -res biomarkers in rat serum and applied this method to pharmacokinetic and stability studies. The mass spectrometer was set to negative ionization mode for the quantification of quercetin and -res. Phenomenex Luna (C18(2), 100 A, 75 × 4.6 mm, 3 µ) column was utilized to separate the analytes using an isocratic mobile phase consisting of methanol and 0.1% formic acid in water (82:18). Validation of the method was performed using various parameters, including linearity, specificity, accuracy, stability, intra-day, inter-day precision, and the matrix effect. There was no observed significant endogenous interference from the blank serum. The analysis was completed within 5.0 min for each run, and the lower limit of quantification was 5 ng/mL. The calibration curves showed a linear range with a high correlation coefficient (r > 0.99). The precision for intra- and inter-day assays showed relative standard deviations from 3.32% to 8.86% and 4.35% to 9.61%, respectively. The analytes in rat serum were stable during bench-top, freeze-thaw, and autosampler (-4 °C) stability studies. After oral administration, the analytes showed rapid absorption but underwent metabolism in rat liver microsomes despite being stable in simulated gastric and intestinal fluids. Intragastric administration resulted in higher absorption of quercetin and -res, with greater C, shorter half-life, and improved elimination. No prior research has been conducted on the oral pharmacokinetics and stability of anti-diabetic compounds in the Ethanolic extract of EECQ, making this the first report. Our findings can provide the knowledge of EECQ's bioanalysis and pharmacokinetic properties which is useful for future clinical trials.
是一种营养丰富的植物，具有传统医学应用的历史。它拥有丰富多样的多酚，包括槲皮素、白藜芦醇、β-谷甾醇、杨梅素和其他化合物。我们开发并验证了一种灵敏的 LC-MS/MS 方法来定量大鼠血清中的槲皮素和白藜芦醇生物标志物，并将该方法应用于药代动力学和稳定性研究。质谱仪设置为负离子模式，用于定量槲皮素和白藜芦醇。 Phenomenex Luna（C18（2），100A，75×4.6mm，3µ）柱用于分离分析物，使用甲醇和水（82:18）中的 0.1%甲酸作为等度流动相。方法验证使用各种参数进行，包括线性、特异性、准确性、稳定性、日内、日间精密度和基质效应。空白血清中没有观察到明显的内源性干扰。每个运行的分析时间为 5.0 分钟，定量下限为 5ng/mL。校准曲线显示线性范围，相关系数高（r＞0.99）。日内和日间测定的精密度分别为 3.32%至 8.86%和 4.35%至 9.61%。大鼠血清中的分析物在台式、冻融和自动进样器（-4°C）稳定性研究中稳定。口服给药后，分析物表现出快速吸收，但在大鼠肝微粒体中发生代谢，尽管在模拟胃液和肠液中稳定。胃内给药可提高槲皮素和白藜芦醇的吸收，增加 Cmax，缩短半衰期，改善消除。在 Ethanolic extract of EECQ 中，没有关于抗糖尿病化合物口服药代动力学和稳定性的先前研究，这是首次报道。我们的发现可以提供 EECQ 的生物分析和药代动力学特性的知识，这对未来的临床试验很有用。
