Liu Junxi, Meng Min, Li Chen, Huang Xinyi, Di Duolong
Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, tianshuizhonglu No. 18, Lanzhou 730000, Gansu, China.
J Chromatogr A. 2008 May 9;1190(1-2):80-5. doi: 10.1016/j.chroma.2008.02.083. Epub 2008 Mar 2.
By optimizing extraction, separation and analytical conditions, a reliable and accurate high-performance liquid chromatographic (HPLC) method coupled with photodiode array detector (DAD) at room temperature is developed for simultaneous determination of three diarylheptanoids (juglanin A, juglanin B, rhoiptelol) and an alpha-tetralone derivative (regiolone) in methanol extracts from the green walnut husks (Juglans regia L.) The sample pretreatment process involved the reflux extraction using methanol as the extract with a ratio of liquor to sample of 15 mL/g. The separation was achieved on a SinoChrom ODS-AP C(18) column with gradient elution using acetonitrile and 2% (v/v) acetic acid in water. The intra-day and inter-day precision (RSD%) for the analytes ranged from 1.08 to 1.51 and 0.60 to 1.13, respectively. The average recoveries obtained were from 88.4% to 96.2% for the analytes with RSDs below 3.13%. The correlation coefficients of the calibration curve exceeded 0.999. The detection limits were 0.51, 0.25, 0.32 and 0.35 ng at a signal-to-noise ratio of 3, respectively. Quantitative analyses of the samples from different grown sites and in obtained different months showed that the contents of the analytes varied significantly. The method was then successfully applied for the detection and isolation of a new diarylheptanoid derivative in the green walnut husks (J. regia L.). The structure of the new compound was elucidated by various spectroscopic methods including 2D NMR techniques (COSY, HMQC, HMBC), HR-ESI-MS and X-ray single-crystal diffraction analysis.
通过优化提取、分离和分析条件，建立了一种可靠、准确的高效液相色谱（HPLC）方法，该方法在室温下结合光电二极管阵列检测器（DAD），用于同时测定绿核桃壳（Juglans regia L.）甲醇提取物中的三种二芳基庚烷类化合物（胡桃素A、胡桃素B、rhoiptelol）和一种α-四氢萘酮衍生物（regiolone）。样品预处理过程包括以甲醇为提取剂，按液料比15 mL/g进行回流提取。采用SinoChrom ODS-AP C(18)柱，以乙腈和2%（v/v）醋酸水溶液进行梯度洗脱实现分离。分析物的日内和日间精密度（RSD%）分别为1.08%至1.51%和0.60%至1.13%。分析物的平均回收率为88.4%至96.2%，RSD低于3.13%。校准曲线的相关系数超过0.999。在信噪比为3时，检测限分别为0.51、0.25、0.32和0.35 ng。对来自不同生长地点和不同月份采集的样品进行定量分析表明，分析物的含量有显著差异。该方法随后成功应用于绿核桃壳（J. regia L.）中一种新的二芳基庚烷类衍生物的检测和分离。通过多种光谱方法，包括二维核磁共振技术（COSY、HMQC、HMBC）、高分辨电喷雾电离质谱（HR-ESI-MS）和X射线单晶衍射分析，阐明了新化合物的结构。
