[논문]Quantitative Analysis for the Quality Evaluation of Scutellariae Radix by HPLC/UVD

Jeong, Su-Yang; Moon, Dong-Cheul; Son, Kun-Ho; Son, Jong-Keun; Min, Byung-Sun; Woo, Mi-Hee

Quantitative Analysis for the Quality Evaluation of Scutellariae Radix by HPLC/UVD 원문보기

Natural product sciences, v.17 no.4, 2011년, pp.321 - 327

Jeong, Su-Yang (College of Pharmacy, Catholic University of Daegu) , Moon, Dong-Cheul (College of Pharmacy, Chungbuk National University) , Son, Kun-Ho (College of Life Science, Andong National University) , Son, Jong-Keun (College of Pharmacy, Yeungnam University) , Min, Byung-Sun (College of Pharmacy, Catholic University of Daegu) , Woo, Mi-Hee (College of Pharmacy, Catholic University of Daegu)

Abstract ▼ AI-Helper

In this study, quantitative analysis was developed using HPLC/UVD for the quality evaluation of Scutellariae Radix. For quantitative analysis, six major bioactive compounds were assessed. The separation conditions employed for HPLC/UVD were optimized using Phenomenex $C_{18}$ column ($250{\times}4.6$ mm, 5 ${\mu}m$) with a gradient of solvent A (1% acetic acid of $H_2O$) and solvent B (acetonitrile : methanol : acetic acid = 70 : 30 : 1) as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 275 nm. These methods were fully validated with respect to linearity, accuracy, precision and recovery. The HPLC/UVD method was applied successfully to the quantification of six major compounds in the extract of Scutellariae Radix. The results indicate that the established HPLC/UVD method is suitable for the quantitative analysis and quality control of multicomponents in Scutellariae Radix.

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제안 방법

Sample analysis − The developed HPLC/UVD method was then applied to the simultaneous determination of the six compounds, 5,7,2',5'-tetrahydroxy-8,6'-dimethoxyflavon (1), baicalin (2), baicalein (3), wogonin (4), chrysin (5) and oroxylin A (6) in the eight Scutellariae Radix samples corresponding to two samples collected from Korea (S1 and S2), four samples cultured in Korea and purchased from markets in Korea (S3, S4, S5 and S6), and two samples cultured in China and purchased from markets in Korea (S7 and S8).
The LOD and LOQ under our experimental conditions are listed in Table 3. The extraction recovery test was performed by extracting a known amount of the six compounds from the Scutellariae Radix powder samples. The % recovery of standard ranged from 98.
For the preparation of the crude extract, the powders of the dried rhizome of Scutellariae Radix were sieved through a 100- mesh. The recovery, precision and accuracy tests were executed by mixing a powdered sample (50 mg) with three control levels (20%, 50%, 100%) of the reference compounds. The mixture was then sonicated with 50 mL of 70% ethanol at room temperature for 60 min.
The sample assay results after the sonication extraction method were higher than those after reflux. To determine the time needed for complete extraction, samples were extracted for five different lengths of time (30, 40, 50, 60 and 70 minutes). Thus 70% ethanol solvent and the sonication extraction method were employed.

대상 데이터

HPLC/UVD conditions − The HPLC equipment was a Waters HPLC system (Waters, 2796) with a Waters 1525 binary pump, an autosampler, a degasser, a column oven and a Waters 2966 photodiode array detector.
Reagents − All of the standard compounds were provided by Prof. K. H. Son, Andong National University, Andong, Korea.

성능/효과

Optimization of sample preparation conditions. Eight extracting solvents, 100% ethanol, 70% ethanol, 50% ethanol, 20% ethanol, 100% methanol, 70% methanol, 50% methanol and 20% methanol, were compared in sample assays after sonication for 60 minutes. When the sample was extracted with 70% ethanol, the sample assay was higher than with the other solvent samples.
The % recovery of standard ranged from 98.32 to 104.12%, and the RSD was less than 1.61% (Table 4).
IX for the determination of baicalin, baicalein and wogonin in Scutellariae Radix. The assay revealed that the total sum of baicalin, baicalein and wogonin of all samples was 13.54 ~ 22.01%, not less than 10.00% (K.P. provision).
12%. The inter-assay precision was lower than 4.75% of RSD and accuracy between 96.71% and 102.54%. The above data reflects that the developed method is highly reproducible; precision and accuracy data are presented in Table 5.
999, as determined by least square analysis, suggesting good linearity between the peak areas and the compound concentrations over a wide concentration range (Table 3). The limits of detection (LOD) and limits of quantitation (LOQ) were evaluated based on the lowest detectable peak in the chromatogram having a signal-to-noise (S/N) ratio of 3 and 10, respectively. The LOD and LOQ under our experimental conditions are listed in Table 3.
Intra-assay precision and accuracy were determined from the variability of triplicate analyses of quality control samples analyzed within the same analytical run. The remaining quality control samples had intra-assay precision below 4.82% of RSD and accuracy between 98.32% and 104.12%. The inter-assay precision was lower than 4.

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