Seo Chang-Seob, Shin Hyeun-Kyoo
K-herb Research Center, Korea Institute of Oriental Medicine, 1672 Yuseong-daero, Yuseong-gu, Daejeon 34054, Korea.
J Chromatogr Sci. 2017 Oct 1;55(9):926-933. doi: 10.1093/chromsci/bmx053.
Yongdamsagan-tang (YDSGT) has been used clinically for the treatment of acute- and chronic-urethritis, cystitis, orchitis and hypertension in Korea. In this study, a powerful method based on high-performance liquid chromatography (HPLC) with photodiode array (PDA) detection was established and validated for the quantitative analysis of eight components: chlorogenic acid, gentiopicroside, liquiritin apioside, liquiritin, nodakenin, baicalin, wogonoside and glycyrrhizin in YDSGT extract. The compounds were separated with a Gemini C18 analytical column (column temperature: 40°C; mobile phase: 0.1% (v/v) aqueous trifluoroacetic acid (A) and acetonitrile (B); flow rate: 1.0 mL/min; injection volume: 10 μL). The PDA detector scanned the range 190-800 nm and the marker compounds were monitored at 254, 275, 325 and 335 nm. The correlation coefficients of all compounds were 1.000 and the results showed excellent linearity. The lower limits of detection and quantification of the analytes were 0.01-0.09 μg/mL and 0.03-0.28 μg/mL, respectively. The extraction recoveries of the marker compounds were 98.13-103.86%, with relative standard deviation values not exceeding 2.10%. The precision of intra- and inter-day measurements were 0.09-1.78% and 0.12-2.09%, respectively. The content of the eight marker compounds in the freeze-dried YDSGT extract were 1.41-23.71 mg/g.
勇丹沙干汤(YDSGT)在韩国临床上已用于治疗急慢性尿道炎、膀胱炎、睾丸炎和高血压。在本研究中,建立了一种基于高效液相色谱(HPLC)和光电二极管阵列(PDA)检测的强大方法,并对勇丹沙干汤提取物中的8种成分进行了定量分析,这8种成分分别为绿原酸、龙胆苦苷、甘草苷、甘草素、紫花前胡苷、黄芩苷、汉黄芩苷和甘草酸。使用Gemini C18分析柱(柱温:40℃;流动相:0.1%(v/v)三氟乙酸水溶液(A)和乙腈(B);流速:1.0 mL/min;进样量:10 μL)对化合物进行分离。PDA检测器扫描范围为190 - 800 nm,标记化合物在254、275、325和335 nm处进行监测。所有化合物的相关系数均为1.000,结果显示出良好的线性关系。分析物的检测限和定量下限分别为0.01 - 0.09 μg/mL和0.03 - 0.28 μg/mL。标记化合物的提取回收率为98.13 - 103.86%,相对标准偏差值不超过2.10%。日内和日间测量的精密度分别为0.09 - 1.78%和0.12 - 2.09%。冻干勇丹沙干汤提取物中8种标记化合物的含量为1.41 - 23.71 mg/g。