HPLC-UVD法测定肿节风水提物及相关制剂中迷迭香酸及异嗪皮啶的含量

Content determination of rosmarinic acid and isofraxidin in Sarcandra glabra extract and preparations by HPLC-UVD

目的采用HPLC法测定肿节风水提取物及相关制剂中迷迭香酸和异嗪皮啶的含量,为肿节风提取物及制剂的质控方法奠定基础。方法采用C18色谱柱（250 mm×4.6 mm,5μm）,以乙腈-水（分别含体积分数为0.1%甲酸）为流动相梯度洗脱,流速1.00 mL.min^-1,检测波长330 nm,柱温35℃。结果迷迭香酸的质量浓度在4.746-189.8 mg.L^-1内、异嗪皮啶的质量浓度在1.018-40.72 mg.L^-1内线性关系良好;仪器精密度试验、重复性试验、稳定性试验RSD均不大于2.0%。肿节风水提取物中迷迭香酸与异嗪皮啶的回收率分别为99.8%和99.2%,RSD分别为0.70%和0.63%（n=6）;迷迭香酸与异嗪皮啶在相关制剂中回收率分别为100.7%和98.4%,RSD分别为1.6%和0.44%（n=6）。12批肿节风水提取物供试品中迷迭香酸的含量在1.479-6.447 mg.g^-1内、异嗪皮啶的含量在1.912-3.540 mg.g^-1内;在以肿节风水提取物制成的相关制剂中,迷迭香酸的含量在0.847-15.946 mg.g^-1内、异嗪皮啶的含量在2.032-4.093 mg.g^-1内。结论作者建立的肿节风及制剂的定量分析方法简便、快速、专属性强、重现性好,为肿节风水提取物及相关制剂质量标准的提升奠定了基础。

Objective To develop an HPLC-UVD method for content assay of rosmarinic acid and isofraxidin in Sarcandra glabra extract and preparations.Methods The method was developed on an C18 column（250 mm×4.6 mm,5 μm） by gradient elution with acetonitrile-water（both containing 0.1% formic acid）as mobile phase at a flow rate of 1.00 mL·min-1 with the wavelength at 330 nm and column temperature at 35 ℃.Results The linearity was good over the range of 4.746-189.8 mg·L-1 for rosmarinic acid and 1.018-40.72 mg·L-1 for isofraxidin.The RSD was no more than 2.0% in precision test,reproducibility test and stability test,respectively.The average recovery of rosmarinic acid and isofraxidin in Sarcandra glabra extract were 99.8%（RSD=0.70%）and 99.2%（RSD=0.63%）,while in preparations were 100.7%（RSD=1.6%）and 98.4%（RSD=0.44%）,respectively.The twelve batches of Sarcandra glabra extract sample shared an average content over the range of 1.479-6.447 mg·g-1 for rosmarinic acid and 1.912-3.540 mg·g-1 for isofraxidin,while 0.847-15.946 mg·g-1 for rosmarinic acid and 2.032-4.093 mg·g-1 for isofraxidin in preparations.Conclusions The method is simple,specific,precise and accurate and can help to improve the quality standard of Herba Sarcandrae extract and preparation.