光果葶苈中莱菔硫烷的提取及含量测定

Extraction Technology and HPLC Determination of Sulforaphane in Draba Nemorosa L.var. Leiocarpa Lindl.

关键词:  光果葶苈  莱菔硫烷  正交试验设计  高效液相色谱法

Key words:  Draba nemorosa L. var. leiocarpa Lindl.  sulforaphane  orthogonal design  HPLC

摘要:

目的 研究十字花科葶苈属植物光果葶苈(Draba nemorosa L. var. leiocarpa Lindl.)中莱菔硫烷的最佳提取工艺，为进一步研究开发此植物奠定基础。方法 利用正交实验设计确定最佳提取工艺，考察因素包括缓冲溶液pH值，超声提取时间和提取温度。使用HPLC检测提取物中莱菔硫烷含量：采用Eclipse XDB-ⅡC₁₈柱，以乙腈和水进行梯度洗脱；梯度洗脱条件：A泵为乙腈，B泵为水；30 min内乙腈的浓度从20%升至50%；检测波长为254 nm；室温。结果 所考察的因素中，对光果葶苈中莱菔硫烷提取率影响程度为：缓冲溶液pH值＞超声提取时间＞提取温度，tris-HCl缓冲溶液的pH对提取率的影响最大；HPLC测定的线性范围为0.45~5.4 µg(r=0.996 8)。结论 光果葶苈中莱菔硫烷最佳提取工艺为：超声提取30 min，tris-HCl缓冲溶液pH值为5.5，提取温度30 ℃；HPLC测定光果葶苈中莱菔硫烷含量的方法快速、准确。

Abstract:

OBJECTIVE To study the extraction technology of sulforaphane from Draba nemorosa L. var. leiocarpa Lindl. to provide scientific data for further exploitation. METHODS The extraction condition was optimized with orthogonal design considering the following factors: the pH of the buffer solution, the time of extraction and the temperature. The HPLC separation was performed on Eclipse XDB-ⅡC₁₈ column with a room column temperature, a mobile phase consisted of acetontrile and water with UV detection wavelength at 254 nm. RESULTS Sulforaphane extraction was affected by pH of tris-HCl, extraction time and temperature. The calibration curve was linear in the range of 0.45-5.4 µg(r=0.996 8). CONCLUSION The optimum extraction condition was as follows: temperature 30 ℃, extract for 30 min with buffer pH 5.5. The sulforaphane from Draba nemorosa L. var. leiocarpa Lindl. could be determined by HPLC on the optimum condition, and this method was rapid and specific,