Study Of The Impact Of Bamboo To The Content Of Aristolochic Acid A Of Caulis Aristolochiae Manshuriensis And Radix Aristolochiae

Objective: This study by means of researching the impact of the Bamboo of different combination proportion on aristolochic acid A in Caulis Aristolochiae Manshuriensis and Radix Aristolochiae and the impact of different Boiling time on aristolochic acid A of the water–cooked-liquid of Caulis Aristolochiae Manshuriensis and Radix Aristolochiae through studying the relationship between quantity and toxicity, the relationship between time and toxicity, to seek the method for controlling even eliminating the quantity of aristolochic acid A in water–cooked-liquid, aimed at reducing aristolochic acid A, to supply dependable proof for clinical application.一. Study of the impact of the Bamboo of different combination proportion on aristolochic acid A in Caulis Aristolochiae Manshuriensis and Radix Aristolochiae.Methods: (1) Qualitative identification: The aristolochic acid A in the water–cooked-liquid of Caulis Aristolochiae Manshuriensis and Radix Aristolochiae is Identificated by TLC (thin-layer chromatography) as follow: TLC conditions: Thin-layer plate: Silica gel G thin-layer plate. Start agent: Toluene:ethyl acetate: formic acid: water =20:10:1:1 skimming,take the upper layer. Detect condition:365nm observed under UV lamp. (2) The aristolochic acid A is analysised by HPLC. Aglient chromatographic column(XDB-C18,4.6×150mm,5μm),The mobile phase:Methanol: water: glacial acetic acid =72:27:1 The detection wavelength:315nm,Temperature:25℃,Flow rate:1ml/min.Results:(1)TLC result:there are same dark spots in Test chemicals for chromatography 1,2,3,4,5 and Reference substance chromatography under 365nm UV lamp,while Pale yellow spot under the light, no spot in No. 6 Negative control group, which shows that the former 5 sample solution contain some aristolochic acid A. (2)HPLC result:when Caulis Aristolochiae Manshuriensis : Bamboo=4:1, the amount of aristolochic acid A is the lowest.二.Study of the impact of boiling time on aristolochic acid A in Caulis Aristolochiae ManshuriensisMethods :( 1 ) The aristolochic acid A in the water–cooked-liquid of Caulis Aristolochiae Manshuriensis is Identificated by TLC as follow: As the toluene: ethyl acetate: formic acid: water (20:10:1:1) Start of. And as the Standard aristolochic acid A Reference to make TLC analysis. (2)The aristolochic acid A is analysised by HPLC as follow: C18 Pole;The mobile phase : Methanol: water: glacial acetic acid (72:27:1);The detection wavelength:315nm;Flow rate:1.0ml/min; Temperature: 25℃. Results:(1)TLC result:The results showed that there is Pale yellow spot on the Plate of The tested Solution at the same Location as that of the Standard Solution under the light. While under the 365nmΜV the spot turn dark. This shows there is some aristolochic acid A in the 2 sample solution in spite of different boiling time(.2)HPLC result:sample 1(boiling time : 45/30min)aristolochic acid A is 0.160 mg/ml,sample 2(boiling time : 30/20min)aristolochic acid A is 0.354 mg/ml.Conclusion:When the Proportion of Bamboo and Caulis Aristolochiae Manshuriensis and Radix Aristolochiae is appropriate, the Bamboo can lower the content of aristolochic acid A of the Plant of Aristolochial. Instead, it can increase the content of aristolochic acid A of the Plant of Aristolochial. The study shows when Bamboo: Caulis Aristolochiae Manshuriensis is 1:4, or Bamboo: Radix Aristolochiae is 1:4; the content of aristolochic acid A is the lowest. So, when Caulis Aristolochiae Manshuriensis and Radix Aristolochiae are used, we can try to add a quarter Bamboo during 45/30min boiling time to lower the content of aristolochic acid A. Under Compatibility Theory of Chinese Medicine, it is possible to reduce the toxicity of Caulis Aristolochiae Manshuriensis and Radix Aristolochiae in proper combination proportion and boiling time.