참깨 잎 추출물 중 pedaliin 함량분석을 위한 HPLC 분석법 밸리데이션

HPLC Method Validation for Quantification of Pedaliin in Sesamum indicum Leaves Extract

Background/Objectives: This study aims to establish the analytical method for quantitative analysis of pedalin in the Sesamum indicum leaves extract (SLE) which has been researched as one of various functional food sources. Methods: SLE and pedaliin were dissolved in methanol and analyzed by high performance liquid chromatography (HPLC) using a reverse phase column (C18, 5 μm, 4.6×250 mm) with a gradient elution of water containing 0.01% trifluoroacetic acid (A) and acetonitrile containing 0.01% trifluoroacetic acid (B): 0 min, 10% B; 5 min, 10% B; 12 min, 20% B; 25 min, 30% B; 30 min, 70% B; 35 min, 90% B; 45 min, 90% B. The eluate was detected at 340 nm using a diode array detector. The column temperature was 30 ℃. The method was validated for specificity, repeatability, linearity, limit of detection, limit of quantitation, accuracy, and precision. Results: Calibration curve for pedaliin showed high linearity with correlation coefficients of 0.9993∼0.9996. Limit of detection and limit of quantitation were 1.310 and 3.971 μg/mL, respectively. In recovery test, the accuracy ranged 98.812∼102.076% with relative standard deviation (RSD) values less than 0.551%. For precision, repeatabilities for retention time and peak area of pedaliin were RSD values of 0.017∼0.025% and 0.119∼1.086%, respectively. In addition, the RSD values of intra- and inter-day tests were 0.124∼2.368%. Conclusion/Implications: The parameters of method validation are reliable for quantitating pedaliin in SLE, and pedaliin can be used as an indicator compound for SLE.