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Article

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

1
Graduate School (in the program of Research and Development in Pharmaceuticals), Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand
2
Division of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand
3
Human High Performance and Health Promotion (HHP&HP) Research Institute, Khon Kaen University, Khon Kaen 40002, Thailand
*
Author to whom correspondence should be addressed.
Molecules 2019, 24(19), 3522; https://doi.org/10.3390/molecules24193522
Received: 8 September 2019 / Revised: 26 September 2019 / Accepted: 27 September 2019 / Published: 28 September 2019
(This article belongs to the Special Issue Selected Papers from the Joint Symposia of MESMAP-5 & ISPBS-5)
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 Sesamum indicum 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 ± 0.008 min) with methanol/water (70%, v/v) through a 30 °C, 5-µm 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 10–1000 ng/mL (R2 = 0.9972). The % accuracy of the sesamol quality control sample was −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 (R2 = 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. View Full-Text
Keywords: Sesamum indicum; Pedaliaceae; sesamol; HPLC; method validation; melanoma; cell-based assay; intracellular concentration Sesamum indicum; Pedaliaceae; sesamol; HPLC; method validation; melanoma; cell-based assay; intracellular concentration
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MDPI and ACS Style

Srisongkram, T.; Weerapreeyakul, N. Validation of Cell-Based Assay for Quantification of Sesamol Uptake and Its Application for Measuring Target Exposure. Molecules 2019, 24, 3522. https://doi.org/10.3390/molecules24193522

AMA Style

Srisongkram T, Weerapreeyakul N. Validation of Cell-Based Assay for Quantification of Sesamol Uptake and Its Application for Measuring Target Exposure. Molecules. 2019; 24(19):3522. https://doi.org/10.3390/molecules24193522

Chicago/Turabian Style

Srisongkram, Tarapong, and Natthida Weerapreeyakul. 2019. "Validation of Cell-Based Assay for Quantification of Sesamol Uptake and Its Application for Measuring Target Exposure" Molecules 24, no. 19: 3522. https://doi.org/10.3390/molecules24193522

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