响铃草药材中总黄酮和3种黄酮苷元的测定方法及含量变化研究

Determination and Content Changes of Total Flavonoids and Three Flavonoid Aglycones in Crotalaria ferruginea Grah

以芦丁和槲皮素、染料木素和山柰素为对照品,对响铃草药材4种黄酮类成分进行含量测定研究。采用紫外分光光度法在500nm波长处测定芦丁的吸光度,以芦丁计得响铃草总黄酮的量,用高效液相色谱法测定响铃草中槲皮素、染料木素和山柰素的含量。生长旺季为3～8月份,4种成分含量均较低;而在当年10月至次年2月（休眠期）,4种成分含量逐渐增高,2月份含量达最高。随着生长年限的增大4种成分的含量升高,1～6年生的植株内3种黄酮苷元含量随着年限的增长而较快增长;6～9年生期间,3种黄酮苷元含量近似直线上升;9年以后增加缓慢;总黄酮含量与年限的增长呈曲线增加,1年生最低,为13.42mg/g;10年生最高,为25.10mg/g,平均每年增加总黄酮1.168mg/g。结果表明,不同生源地的响铃草中,总黄酮、槲皮素、染料木素和山柰素4类成分含量最高的生源地为四川邻水长安乡,云南昆明和云南宾江总黄酮和槲皮素差异不大,但都低于四川邻水长安乡,只是云南宾江的染料木素和山柰素含量略高于云南昆明;4种成分含量最低的为贵州雷山县。

Rutin, quercetin, genistein and kaempferol were used as reference substances. Their contents were determinated in Crotalaria ferruginea Grah. Ultraviolet spectrophotometry was used to determine absorbance of rutin at 500 nm wavelength. Thus total flavonoids were inspected. The contents of quercetin, genistein and kaempferol were determined by HPLC from C.ferruginea. The results were as follows ： In the growing season （ from March to August）, the contents of the 4 components were all low, but in the dormancy（ October to the following February）, the contents of the four components gradually increased. The contents in the following February was the highest. The contents in the samples from different growth years increasd with the growth period. In the plants which had grown from 1 to 6 years, the contents of the three flavonoid aglycones increased with increasing age. In the plants which had grown from 6 to 9 years, the contents had a linear ascending tendency. After the plant grew more than 9 years, the contents increased slower. In the samples from diffenert growth years, the content of flavonoids was also increased with increasing age. But the difference was that in the whole process, the content of flavonoids had a curved change. The content of 5-year old plant was the lowest（ 13.42 mg/g）, and 10-year old plant was the highest（25.10 mg/g）, the average annual increase was 1. 168 mg/g. The results showed that the hightest contents of total flavonoids, quercetin, genistein and kaempferol were in C.ferruginea from Linshui Chang＇an of Sichuan Province. The contents of total flavonoids and quereetin in C. ferruginea from Binjiang and Kunming of Yunnan Province had little difference and they were less than those of Chang＇an. Only the contents of genistein and kaempferol in C. ferruginea from Binjiang were more than in Kunming. The lowest contents of total flavonoids, quercetin, genistein and kaempferol were in C.ferruginea from Leishan County, Guizhou Province.