| Objective:The dissertation is conducted to study the principles of compatibility "reinforcing yang from yin" of kidney-qi pill. Through the establishment of stable, reliable conditions of high performance liquid chromatogram(HPLC), the study detects the active ingredients cinnamaldehyde,paeonol and loganin.Then the principles of compatibility "reinforcing yang from yin" is analyzed in accordance with its change. In addition, through the analysis of the difference of spectrum of chemical components in each group in vitro and in vivo, the investigation lays the root for the following research of colour spectral changes for kidney-qi pill in pharmacokinetic.Method:By exploring the chromatographic condition such as mobile phase,wave length of detection etc, this study establishes the technology of active component detection. According to the feature of "reinforcing yang from yin" of kidney-qi pill, the kidney-qi pill is split into groups associated with the "three-reinforce" and "hree-decanta". According to the established chromatographic conditions, the study measures the active component Cinnamaldehyde, paeonol and loganin. Then the peak area is brought into comparisons to analyze the impact of compatibility for the three active component. Then the study continues to detect blood serum samples which gain from the lavaged rat of each drug and group, and compare the changes of color spectrum. After tracing back the migrated components, drug resource and metabolic product in blood serum of rat, the principles of compatibility "reinforcing yang from yin" for kidney-qi pill is initially elaborated in this dissertation.Result:The chromatographic conditions of our study show as fallows:chromatographic column Platisi ODS (5μm,250×4.6mm); mobile phase:gradient elution with acetonitrile-1% glacial acetic acid. flow rate 0.5ml/min, wavelengt 254nm; column temperature 25℃, sample size:5μl.The chromatographic condition of detection with good stability,precision and reproducibility.The results of each split kidney-qi pill group in this chromatographic condition is as follows:The peaks of number 8,22,26,29,32-33,35,37-38,48,50,52-53,55,58,60 are from moutan cortex; the peaks of number 15,19-21,23,25,28,31,40,43 are derived from asiatic cornelian cherry fruit; peaks of number 14,34 come from rehmannia;and number 28,32,38,54 peaks from ramulus cinnamomi.There was no peak from hoelen, alismarhizome,dioscoea root and aconite root. Peaks of number 13,17-18,24,27,30,36,39,42,44-47 come from Moutan cortex and asiatic cornelian cherry fruit in common;peaks of 11,49 are from Moutan cortex and rehmannia in common;Peaks of 41,51 are derived from moutan cortex,rehmannia and ramulus cinnamomi in common.It was found that the peaks areas of cinnamaldehyde in decotion of kidney-qi pill group "reinforcing yang from yin" are higher than invigorating ying-yang group,invigorating yang group,ramulus cinnamomi single group. While the peasks areas of invigorating ying-yang group are higher than invigorating yang group, and promote secretion of cinnamaldehyde, the better of compatibility with hoelen,alisma rhizome,moutan cortex.The peak areas of loganin in the invigorating ying-yang groups are higher than the shenqi pill group"reinforcing yang from yin".But the body's serum of kidney-qi pill group"reinforcing yang from yin"reduced mostly.The peaks areas of paeonol in kidney-qi pill compound group are obviously increased.The experimental set up HPLC of kidney-qi pill serum, then analysis the compounds absorbed into blood and its metabolites of kidney-qi pill. And the results are displaied as follows.In the analysis colour spectrums of rehmannia, the number of 20,21,23 peaks are derived from the originated composition of rehmannia,and the peaks of number 28,37,38,39 are metabolites.In the analysis colour spectrums of dioscorea root, we speculate that the number of 21,29,31 peaks are metabolites,number 21 of which also derived from rehmannia.In the analysis colour spectrums of hoelen, we speculate that the number of 20,21,26,29 peaks are metabolites, the number of 26,29 peaks of which are also derived from the originated composition of kidney-qi pill group, but not in the hoelen single group and we can infer that hoelen may be the effective constituent after absorbed into blood.In the analysis colour spectrums of alisma rhizome, there is nothing could be detected.Conclusion:Ramulus cinnamomi, aconite root compatibility with rehmannia, asiatic cornelian cherry fruit and dioscorea root could increase the dissolution of ramulus cinnamomi, and promote secretion of cinnamaldehyde, the better of compatibility with hoelen,alisma rhizome,moutan cortex. Thus it embodies the mechanism of compatibility of shenqi pill "less fire gas generation". we deduce that it may generate new metabolites after compatibility with rehmannia,asiatic cornelian cherry fruit,and dioscorea root, while the peaks areas of cinnamaldehyde in shenqi pill compound group are rising, which enhance it in blood-vessel penetration after compatibility with hoelen,alisma rhizome,moutan cortex,thus embodies the necessity and scientificity.Compared with invigorating ying-yang group, the peaks areas of loganin in kidney-qi pill compound group are decreased. On one hand, it exist mutual transformation of rehmannia,asiatic cornelian cherry fruit,and dioscorea root, therefore it can promote the dissolution of chemical ingredient. On the other hand, it can affect the dissolution after compatibility with hoelen,alisma rhizome,moutan cortex but lower.The peaks areas of paeonol in kidney-qi pill compound group are obviously increased, thus we deduce rehmannia,asiatic cornelian cherry fruit and dioscorea root which could help to dissolute the moutan cortex and enhance it in blood concentration. Paeonol is likely to be the effective ingredient of kidney-qi pill "reinforcing yang from yin" in blood concentration.In this study, we make primary research on the attribution of the chemicals of kidney-qi Pill in the blood, which may provide evidence to the further research of pharmacokinetic chemical spectrum. |
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