HPLC测定决明不同组织中大黄酚和橙黄决明素含量

摘要

为了测定决明不同组织中大黄酚和橙黄决明素的含量.分别收集了决明种子、根、茎、花、叶、果荚、子叶、胚轴、愈伤与不定根等组织,利用回流提取法提取大黄酚与橙黄决明素,高效液相色谱法(HPLC)进行含量测定.采用依利特反相色谱柱,以大黄酚和橙黄决明素为指标,以乙腈-0.1％磷酸水为流动相进行梯度洗脱,流速1 mL/min,检测波长为284 nm,柱温30℃.结果表明:大黄素型蒽醌类化合物在决明中的分布具有明显的组织特异性,其中,决明种子、胚轴、不定根、子叶与愈伤组织中含有大黄酚与橙黄决明素,决明根中仅含有大黄酚,其余组织不含有大黄酚与橙黄决明素,其余总蒽醌含量依次为子叶＞不定根＞愈伤组织＞胚轴＞根＞花.决明种子成熟过程中,大黄酚的含量呈现“波浪式上升”的趋势,峰值出现在11月下旬;而橙黄决明素的含量呈现“先上升后下降”的趋势,峰值出现在11月中旬.由于大黄酚与橙黄决明素的含量在决明不同成熟时期分别出现峰值,建议采收11月下旬的决明种子以获得大黄酚,采收11月中上旬的决明种子以获得较多的橙黄决明素.%In order to determine the content of chrysophanol and aurantio obtusin in different tissues of Cassia tora,the seed,root,stem,flower,leaf,pod,cotyledons,hypocotyl,callus and adventitious root were collected,the chrysophanol and aurantio obtusin were extracted using reflux extraction.The content of chrysophanol and aurantio obtusin was determined using HPLC method with Hypersil ODS2 column (4.6 mm × 200 mm,5 μm).The analysis condition was as follows:flow rate of 1 mL/min,detection wavelength at 284 nm,temperature at 30 ℃,acetonitrile and 0.1％ phosphoric acid in water as mobile phase.The results showed that,the distribution of chrysophanol and aurantio obtusin in C.tora was tissue-specific.The seed,hypocotyl,cotyledon,adventitious roots and callus contained aurantio obtusin and chrysophanol.The root contained chrysophanol merely and the other tissues did not contain either chrysophanol or aurantio obtusin.The total anthraquinone content was cotyledon ＞ adventitious root ＞ callus ＞ hypocotyl ＞ root ＞ flower.During the mature process of seed,the content of chrysophanol showed wave-like increasing tendency with a peak value appeared in the end of November and the content of aurantio obtusin showed bell-shape curve with a peak value appeared in the beginning of November.As the content of chrysophanol and aurantio obtusin in the different maturity of different peak,it is recommended to harvest in late November of the cassia seed to obtain chrysophanol,harvest in early November of the cassia seed to get more aurantio obtusin.The total anthraquinone,chrysophanol and aurantio obtusin were the highest in the mature cassia seed,indicating that the medicinal value of the part was the highest,hence it was scientific to choose the seed.In our country,Cassia has always been the seed medicine,but the collection of seed due to seasonal restrictions,can only be collected once a year,and the number is limited.Plant tissue culture of callus,adventitious root are not subject to seasonal restrictions,and easy to collect large amount,it is recommended to carry out large-scale tissue culture as a new anthraquinone source.The trend of accumulation of chrysophanol and aurantio obtusin in different tissues of Cassia was reported in this study.It provided information for the further study on the expression and regulation of related genes in the synthesis process of chrysophanol and aurantio obtusin.