Comparison Study On Alkaloid And Anti-platelet Aggregation Between Chinese Chuanxiong And Japanese Chuanxiong

Both Chinese Chuanxiong (CCX) and Japanese Chuanxiong (JCX) have the functions of facilitating blood circulation and dispersing blood stasis. In traditional Chinese medicine, ethnomedicines of Korean nation and Japanese-Chinese medical, CCX and JCX have the same name and uses, used as "Chuanxiong" for treating headaches, gynecological diseases and so on. JCX easily confused with CCX on account of cultivating in China. According to other literatures, tetramethylpyrazine (TMP) is the active component of CCX. And the functions of facilitating blood circulation and dispersing blood stasis can been reflected by the anti-platelet aggregation. Therefore, the alkaloids and anti-platelet aggregation of CCX and JCX were studied in order to provide the basis for both the quality assessment and rational drug use.Firstly, comparison of TMP:After the preparation methods and chromatographic conditions have been optimized, the method of quantitative detection of TMP in CCX and JCX samples has been established. The scanning of TMP in 26 CCX samples and 11 JCX samples showed that only 6 CCX samples and 5 JCX samples found TMP (mean of CCX and JCX are 2.75,1.52 μg·g-1). Therefore, the content of TMP in CCX and JCX herbs were low, had no significant difference. TMP is not the main bioactive compound in CCX and JCX, and not suitable for using as the index component of their quality evaluation.Secondly, comparison of total alkaloids:After the various factors been investigated, the optimum conditions are as follows:total alkaloids extracted with hydrochloric acid-ethanol. The acid dye was bromocresol green with the amount of 7.0 mL. The pH of buffer solution was 3.6, which the consumption was 4.0 mL. After extracted for 3 times,1.00 g of anhydrous sodium sulfate was used to removal of water for 30 min. And the detection wavelength was 414 nm. And the acid dye colorimetry method was established to determine total alkaloids in CCX and JCX. As a result, the total alkaloids content of CCX (mean:0.524%, n= 13) was higher than JCX (mean: 0.372%, n= 6), which shows the quality of CCX was better than JCX in terms of total alkaloids. The samples from different sources contain different amount of total alkaloids. The total alkaloids content of the samples collected from production place (mean:0.645%, n= 9) was significantly higher than that the samples bought from the market of herbal medicine (mean:0.253%, n= 4). It’s may be the result of different producing area and storage time of the samples bought from the market of herbal medicine. Furthermore, the mean content of total alkaloids collected in different places show that CCX of Dujiangyan (0.806%, n= 3)> Meishan (0.658%, n= 2)> Pengzhou (0.542%, n= 3). It suggests that there e might be a correlation between the content of total alkaloids and production place.Thirdly, comparison of infrared spectral (IR) fingerprints:IR was applied to compare the chemical compositions of CCX and JCX. The results showed that the peak shape of infrared spectrogram of CCX and JCX was similar. Both of them have the characteristic peak of esters (2928,1743,1647 cm-1). But the correlation coefficients of CCX were greater than JCX. Furthermore, the Sceond-order derivative spectrum at 1746,1639,1079,1015 and 868 cm-1 also can be identified CCX and JCX. It shows that CCX and JCX have the similar chemical compositions, but also have difference. In addition, there is big difference in the correlation coefficients and infrared absorption at 1200-950 cm-1 between the original medicinal and the processed of JCX. It may be the result of variation of chemical during the processing.Fourthly, comparison of anti-platelet aggregation:After the various factors been investigated, the optimum conditions are as follows:blood was collected from rabbit heart and centrifuged to obtain platelet-rich plasma (PRP) and Platelet-poor plasma (PPP). PRP was placed at room temperature and the final platelet count was adjusted to 200 ×109/L by dilution with PPP. The amount of ADP (500 μmol·L-1) was 10 uL and the platelets were utilized within 4 h. And a method which can accurately determined platelet aggregation induced by adenosine diphosphate (ADP) in vitro was established. The results show that there was a dose-effect relationship between the concentration of samples and platelet inhibiting rate. And the platelet inhibiting rate of different extracts shows the 70% ethanol extract> the mixture of alcohol extract and water extract> ethanol extract> water extract> the water extract after alcohol extract. The platelet inhibiting rate of 3 CCX samples and 3 JCX samples show that the effect of JCX on platelet aggregation is as good as CCX, with the inhibition rate of 1 mg crude drug was 29.0%(n= 3) and 29.5% (n= 3), respectively. Therefore, the quality of CCX and JCX had no significant difference by evalution of the inhibition on platelet inhibition aggregation.The chemical compositions of CCX and JCX are similar. And they have no significant difference in the content of TMP and anti-platelet aggregation. But the total alkaloids content of CCX was higher than JCX, and it was the active component of Chuanxiong. Thus, the quality of CCX was superior to JCX.Finally, this paper provides an overview of the growth environment, cultivation and harvesting, identification, chemical composition and pharmacological effects.