Study On The Maturity Of Sogisu And Its Partial Pharmacology

Sanqi is the root of Panax notoginseng (Burk.) F. H. Chen, Ginseng family, with warm-natured, sweet and a little bitter in taste, liver and stomach channels. It has the activities of scattered stasis and hemostasis, swelling analgesia, used to cure diseases such as hemoptysis, hematemesis, epistaxis, hematochezia, metrorrhagia, bleeding wound, chest and abdominal sticking pain, swelling or flutter. Processed Panax notoginseng can restore vital energy, enrich the blood, tone up the system, used as edible drug to cure syndrome of blood deficiency, postpartum weakness and insufficiency of vital energy and blood principally, and also to improve weakness with pale, dizziness, limb weakness, loss of appetite.At present, there has been many studies on raw Panax notoginseng, but just a few on processed notoginseng. As a commonly used Chinese medicine, only raw Panax notoginseng has been noted in Chinese Pharmacopoeia2010edition. The studies on processing method of Panax notoginseng, high pressure steaming is common, but on normal pressure steaming, frying or else are not. This experiment did a lot on quantitative determination, processing, fingerprint, pharmacological study of Panax notoginseng, to elaborates systematically components and pharmacological differences before and after processing, which would lay the foundation for the research of processing mechanism of Panax notoginseng. The research methods and results are as follows:1Contents determination of5saponins and dencichine in Panax notoginseng1.1Contents determination of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rd in Panax notoginseng by HPLC simultaneouslyThe separation was performed on Agilent Zorbax SB-C18column (4.6mm×250mm,5μm), with elution gradient (water:acetonitrile):0～21min80:20,21～30min80:20～75:25,30～41min75:25～59:41,41～-51min59:41～55:45,51～56min55:45～80:20,56～66min80:20; column temperature:20℃, sample size:10μL, flow rate:1mL·min-1and the wavelength of UV detector:203nm. With this method, linear equations of5compounds were notoginsenoside R1Y=304.60X-1.1457, ginsenoside Rg1Y=371.97X-29.646, ginsenoside Re Y=391.18X-6.7571, ginsenoside Rb1Y=270.76X+1.0893, ginsenoside Rd Y=329.50X+1.3643(r=0.9997～0.9998) and recovery ranges were97.6%-101.7%. 1.2Content determination of dencichine in Panax notoginseng by HPLCThe separation was performed on Agilent Zorbax SB-Aq column (4.6mmx250mm,5μm), with elution ratio:35:75(methanol:0.05%glacial acetic acid), column temperature:20℃, wavelength of UV detector:213nm, flow rate:0.8mL·min-1, sample size:10μL. With this method, linear equations was Y=1980.4X-83.431(r=0.9999).Through investigating the contents of dencichine in raw and steamed Panax notoginseng, we found that the percentage was0.59%in raw and0.29%in processed.2Processing studies of Panax notoginseng2.1Processing procedure study of Panax notoginsengInvestigating four factors, moistening temperature and time, drying temperature, steaming time in processing. It was decided that moistening temperature was60℃, moistening time was24-27h, drying temperature was40℃, steaming time was6h.2.2Heat processing technology study of Panax notoginsengThe contents of five saponins in Panax notoginseng before and after steaming, frying, stir-baking with sand were investigated. By HPLC, the contents of notoginsenoside Ri, ginsenoside Rgi.ginsenoside Re, ginsenoside Rb1, ginsenoside Rd were0.98%,3.27%,0.38%,2.72%,0.66%respectively in raw samples; steamed samples0.79%,2.69%,0.33%,2.49%,0.61%; fried samples1.09%,2.56%,0.18%,2.44%,0.57%; stir-baked with sand samples0.30%,1.15%,0.11%,1.00%,0.24%.Through comparing the chromatograms before and after processing, we found that the contents of five saponins decreased and new components were produced after processing. Change extent of both were relative with the heat temperature.3Fingerprint study on saponins of Panax notoginseng3.1Establishment and Method validation of the fingerprint of Panax notoginsengThe influences of extraction method and elution gradient on fingerprint were investigated. Chromatographic conditions were finally determined that Agilent Zorbax SB-C18column (4.6mm×250mm,5μm), with elution gradient (water:acetonitrile):0～25min80:20,25～30min80:20～75:25,30～41min75:25～59:41,41～51min59:41～55:45,51～56min55:45～80:20,56～70min 80:20; column temperature:20℃, sample size:10μL, flow rate:1mL·min-1, and wavelength of UV detector:203nm. With this method, the relative standard deviation (RSD) of instrument precision, repeatability, stability were all less than3%.3.2Study on fingerprints of steamed Panax notoginseng with different steaming timeSteam the samples Oh,2h,4h,6h,8h separately, test the corresponding fingerprints, marking11common peaks. The similarity degrees of Oh,2h,4h>0.99, of which6h,8h>0.76(6h<8h). From the cluster analysis results, Oh,2h,4h,8h were bracketed together and6h was classed to another category. The similarity degrees and the cluster analysis results showed that6h fingerprint had the biggest difference with Oh,2h,4h,8hs’. There is a case for6h appropriate steaming time, which is consistent with the result of processing procedure study.4Comparative study on partial pharmacological effects of raw and steamed Panax Notoginseng4.1Comparison of hemostatic effects between raw and steamed Panax notoginsengWith tail cutting and glass capillary tube method, bleeding time and coagulation time of mice were tested. Medication administration teams all could significantly short blooding time. Raw Panax notoginseng could significantly short coagulation time when the steamed ones had the tendency to reduce the blood coagulation time but not significant. It can be concluded that raw Panax notoginseng has better hemostasis activity.4.2Efficacy comparison of activating blood between raw and steamed Panax notoginseng6%high molecular dextran (iv.) and ice water were used to cause mices microcirculation disturbance. Microcirculation was observed and the blood shear rate was measured to evaluate the activating blood effects of raw and processed Panax notoginseng. Raw and steamed Panax notoginseng had no significant effect on artery width and also on vein width at10min,20min after administration. High and low dose of raw Panax notoginseng both could increase vein width and the number of dilatation of capillary at30min and the number of dilatation of capillary at20min.It shows that raw Panax notoginseng is superior to steamed samples on improving microcirculation effects, of which high dose group has certain effects but effects slow; on promoting blood viscosity, steamed Panax notoginseng is better than the raw one. 4.3Efficacy comparison of enriching blood between raw and steamed Panax notoginsengThe experiment was performed on mices artificially induced acute hemorrhagic anemia. The blood routine indexes were tested to evaluate the promoting effects of raw and steamed Panax notoginseng on blood loss anemia mices. Raw and steamed Panax notoginseng both could significantly increase Red Blood Cell (RBC) number, Hemoglobin (Hb) content and White Blood Cell (WBC) number of blood loss anemia mices. Steamed groups did slightly better, but there were no significant differences between raw and steamed. No significant influence was performed on immune organ weights of blood loss anemia mices. Raw and steamed Panax notoginseng both can significantly tonify blood for blood loss anemia mices and steamed samples do slightly better, but there are no significant differences.4.4Efficacy comparison of tonifying deficiency between raw and steamed Panax notoginsengCyclophosphamide was injected to induce mice Qi deficiency. The blood routine indexes and immune organ (thymus, spleen, adrenal gland) weights were determined to observe the tonifying deficiency effects of raw and steamed Panax notoginseng. Raw and steamed Panax notoginseng both had no significant influence on RBC number of blood deficiency mices induced by cyclophosphamide. Compared with the model group, high dose group of steamed Panax notoginseng could significantly increase the Hb content of blood deficiency mices. All medication administration teams significantly increased the WBC number in blood deficiency mice, especially compound E-Jiao slurry group and high dose of steamed Panax notoginseng group. The immune organ (thymus, spleen, adrenal gland) indexes of high and low dose steamed Panax notoginseng group were quite significantly higher than model group. That of the high and low dose of raw Panax notoginseng increased, but the effects were not significant. It can be seen that steamed Panax notoginseng performs better on improving immunity and BuQi activities; raw Panax notoginseng has better hemostasis activity.