Validation of Cell-Based Assay for Quantification of Sesamol Uptake and Its Application for Measuring Target Exposure

• Main Authors: Tarapong Srisongkram, Natthida Weerapreeyakul
• Format: Article
• Language: English
• Published: MDPI AG 2019-09-01
• Series: Molecules
• Subjects: <i>sesamum indicum</i>; pedaliaceae; sesamol; hplc; method validation; melanoma; cell-based assay; intracellular concentration
• Online Access: https://www.mdpi.com/1420-3049/24/19/3522

The intracellular drug concentration is needed for determination of target exposure at the site of action regarding its pharmacological action and adverse effects. Sesamol is an antiproliferative molecule from <i>Sesamum indicum</i> with promising health benefits. We present a method for measuring the intracellular sesamol content using reverse-phase HPLC with a UV diode array in melanoma cells. Sesamol was completely resolved by isocratic elution (4.152 &#177; 0.008 min) with methanol/water (70%, <i>v</i>/<i>v</i>) through a 30 &#176;C, 5-&#181;m C-18 column and detection at 297 nm. The present assay offers high sensitivity, fast elution, and an accurate and linear nominal concentration range of 10&#8722;1000 ng/mL (<i>R</i>² = 0.9972). The % accuracy of the sesamol quality control sample was &#8722;3.36% to 1.50% (bias) with a 0.84% to 5.28% relative standard deviation (RSD), representing high repeatability and high reproducibility. The % recovery was 94.80% to 99.29%, which determined that there was no loss of sesamol content during the sample preparation. The validated method was applied to monitor intracellular sesamol concentration after treatment from 5 min to 24 h. The remaining intracellular sesamol content was correlated with its antiproliferative effect (<i>R</i>² = 0.9483). In conclusion, this assay demonstrated low manipulation, quick elution, and high sensitivity, precision, accuracy, and recovery, and it was successfully applied to the quantification of sesamol in target cells.