| This dissertation includes five parts:Part one:summary of the related literaturesThis chapter reviews the research ideas,research methods and applications of the compatibility of traditional Chinese medicine compound(TCMC),and forecasts the compatibility study of TCMC.Meanwhile,a total of 633 chemical components(part of the volatile components)from Simiao Yong’an Decoction(SYD)and its component herbs are summarized.188 literatures are cited in this chapter.Part two:serum chemistry study of Simiao Yong’an DecoctionFourteen migrating components absorbed into blood are identified through the LCMS-IT-TOF technology,including nine prototype components and five metabolites.The medicated serum of the total ion current chromatograms of SYD and its component herbs are compared to confirm the origin of every serum components.Component 2,loganin(4),sweroside(5),hyperoside(7)and isochlorogenic acid C(11)are from Lonicera Japonica(principal);Harpagide(1),angoroside C(10)and harpagoside(13)are from Radix Scrophulariae(assistant).Sulfates and glucuronides of liquirtigenin(8,12,14),and liquiritin(6)are from Licorice(mediator).Chlorogenic acid(3)is from both Lonicera Japonica(principal)and Angelicae Sinensis Radix(complement).By comparing the total ion current chromatograms of the sample solution in vitro of SYD and its component herbs to the medicated serum of SYD,harpagide(1),chlorogenic acid(3),loganin(4),sweroside(5),liquiritin(6),hyperoside(7),angoroside C(10),isochlorogenic acid C(11)and harpagoside(13)are confirmed to be prototype components.By comparing the total ion current chromatograms of the medicated serum of SYD to its component herbs,component 2,component 9 and sulfates and glucuronides of liquirtigenin(8,12,14)are confirmed to be metabolites.In the following study,the biological activities of the components absorbed into blood can be further investigated to illuminate the pharmacodynamics material basis in vivo of SYD,and it is suitable for quality control of SYD and its component herbs.Part three:study on the compatibility of Simiao Yong’an Decoction based on prototype components into the blood by decoction modelThe content determination method for the nine components absorbed into blood of water decoction of SYD using UPLC-MSn in negative ion mode technology is established.The quantitative ions are as follows:harpagide(m/z 183.06),chlorogenic acid(m/z 191.04),sweroside(m/z 125.00),loganin(m/z 227.03),liquiritin(m/z 135.02),hyperoside(300.07),isochlorogenic acid C(m/z 353.11),angoroside C(m/z 175.05),harpagoside(m/z 345.13).The corresponding regression equations are Y = 13.6102 X + 1555.7450(r = 0.9995),Y =85.9195 X+ 74658.0895(r = 0.9994),Y= 6.7546 X+401.3286(r= 0.9998),Y= 27.4063 X +89.7270(r= 0.9990),Y= 570.4144 X+ 4628.2593(r= 1.0000),Y= 356.9293 X-825.7706(r = 0.9996),Y = 93.8354 X+ 7178.6005(r = 0.9993),Y = 218.7902 X + 4923.2821(r ＝0.9992),Y= 135.1618 X-130.4727(r = 1.0000),respectively.Each component shows good linear relationship in the individual range of 139.156-10095.587,683.126-24962.377,56.881-4126.646,10.019-726.861,54.441-3949.616,4.190-304.014,115.297-8364.659,37.065-1354.404,10.816-784.711 μg·g-1,and their contents are 0.1977%,0.8555%,0.1317%,0.0355%,0.1496%,0.0035%,0.1655%,0.0524%,0.0440%(n = 6),respectively.The average recoverys are from 84.125%to 113.376%,and the RSD is less equal than 10.265%.The established approach has high accuracy,sensitivity and stability that can be used in the separated prescription experiments.The extraction ratios(ER)for the nine components absorbed into blood of SYD and its different separated prescriptions are detected.The ER of chlorogenic acid in Lonicera Japonica could all be improved(8.53%-12.00%)by two drugs compatibility groups(DCGs)or reduced(8.99%-13.74%)by three drugs compatibility groups.The ER of chlorogenic acid in SYD is obviously lower(8.97%-12.44%)than single or two DCGs,however slightly higher(8.55%-13.30%)than three DCGs.The ER of isochlorogenic acid C in Lonicera Japonica could all be reduced(2.09%-66.48%)by two or else three DCGs while that in SYD is higher than any DCG.The ER of sweroside in Lonicera Japonica improves(1.27%)after compatibility with Angelicae Sinensis Radix,while other DCGs all reduces its ER.Different DCGs all reduce(41.30%-90.78%)the ER of loganin in Lonicera Japonica.The ER of hyperoside in Lonicera Japonica improves(2.44%-13.38%)after compatibility with Angelicae Sinensis Radix,Licorice or both of them.The ER of hyperoside in SYD is distinctly lower(2.62%-18.59%)than any other DCGs.The ER of harpagide in Radix Scrophulariae is lower(9.82%-99.78%)than other different DCGs,thereinto the ER in SYD is the lowest(49.40%).Different DCGs could all reduce(22.76%-42.87%)the ER of angoroside C in Radix Scrophulariae.The reducing effect of three DCGs is apparently higher than that of the two DCGs.Different DCGs could all reduce(7.46%-83.74%)the ER of harpagoside in Radix Scrophulariae.The reducing effect of two DCGs is apparently higher than that of the three DCGs.Different DCGs could all reduce(51.02%-93.03%)the ER of liquiritin in Licorice.After compatibility with Lonicera Japonica Licorice and Radix Scrophulariae,the ER of liquiritin in Licorice comes to the lowest.Moreover,the inhibiting effect of the ER for liquiritin in three DCGs is slightly higher than that in two DCGs.Part four:study on the compatibility of Simiao Yong’an Decoction based on prototype components into the blood by in vivo intestinal absorption modelThe in vivo intestinal absorption content determination method for the nine components absorbed into blood of SYD using UPLC-MSn in negative ion mode technology is established.The quantitative ions are as follows:harpagide(m/z 183.11),chlorogenic acid(m/z 191.05),sweroside(m/z 125.07),loganin(m/z 227.12),liquiritin(m/z 255.13),hyperoside(300.20),isochlorogenic acid C(m/z 353.18),angoroside C(m/z 175.05),harpagoside(m/z 147.03).The corresponding regression equations are Y = 13.4291 X + 1855.7552(r = 0.9993)、Y =91.4800 X+50080.5633(r = 0.9992)、Y=6.9746 X+ 180.1241(r = 0.9999)、Y=30.1934 X-184.5150(r = 0.9995)、Y = 568.2259 X + 5623.1312(r = 1.0000)、Y = 351.2752 X －489.7681(r= 0.9998)、Y=94.0494 X 8443.4213(r = 0.9996)、Y 220.0985 X + 5120.6526(r = 0.9991)、Y = 134.0560 X + 36.0185(r = 1.0000),respectively.Each component shows good linear relationship in the individual range of 138.857-10073.892、681.647-24908.336、56.758-4117.722、9.954-722.159、54.375-3944.814、4.181-303.358、115.047-8346.553、36.985-1351.469、10.793-783.013 μg·g-1.The contents of low dose group are 0.2162%,0.9211%,0.1357%,0.0261%,0.1550%,0.0036%,0.1504%,0.0518%,0.0456%(n=6),respectively.The average recoverys of low dose group are from 83.834%to 112.324%,and the RSD is from 4.850%to 14.141%.The contents of middle dose group are 0.1971%,0.8676%,0.1285%,0.0228%,0.1497%,0.0036%,0.1756%,0.0582%,0.0444%(n = 6),respectively.The average recoverys of middle dose group are from 84.524%to 117.079%,and the RSD is from 1.639%to 13.814%.The contents of high dose group are 0.1968%,0.8895%,0.1318%,0.0223%,0.1485%,0.0036%,0.1883%,0.0596%,0.0441%(n = 6),respectively.The average recoverys of high dose group are from 85.176%to 119.501%,and the RSD is from 2.102%to 7.149%.The established approach has high accuracy,sensitivity and stability,and can be used to determine the contents of the components absorbed into blood of SYD.The rats’ absorption rate constants(Ka)and apparent permeability coefficients(Papp)in duodenum,jejunum,ileum and colon are compared.Duodenum and ileum are determined as the point of penetration of the followed experiments.Duodenum:The intestinal absorption parameters(Ka,Papp)values of chlorogenic acid in SYD are higher than any compatibility combination.Angelicae Sinensis Radix can increase the absorption of chlorogenic acid in the duodenum,nevertheless Licorice shows the opposite effect.The parameter values of sweroside and loganin are higher in SYD or Lonicera Japonica than in other DCGs,but the statistical analysis does not have significant difference.The parameter values of hyperoside in Lonicera Japonica are the highest(P<0.01)after compatibility with Licorice.The parameter values of isochlorogenic acid C in Lonicera Japonica are higher(P<0.05)in SYD or Lonicera Japonica,the parameter values come to the highest(P<0.01)after Lonicera Japonica compatibility with Radix Scrophulariae and Licorice.The parameter values of harpagide in SYD and Angelicae Sinensis Radix lacked group are significantly higher(P<0.01)than other DCGs.In terms of angoroside C,its intestinal absorption parameter values are the lowest(P<0.05)in individual Radix Scrophulariae group.The parameter values of harpagoside are the highest in SYD,but the lowest in Angelicae Sinensis Radix.The parameter values of liquiritin in Licorice are higher(P<0.05)than any other DCGs after Licorice compatibility with Angelicae Sinensis Radix.Ileum:The intestinal absorption parameters(Ka,Papp)values of chlorogenic acid in SYD are the highest.Angelicae Sinensis Radix can increase the absorption of chlorogenic acid in the ileum,nevertheless Licorice shows the opposite effect.The parameters values of sweroside and loganin are lower in SYD and Lonicera Japonica than that in other DCGs.Licorice could inhibit the intestinal absorption of hyperoside in ileum,while the parameters values of hyperoside are in the higher level after Lonicera Japonica compatibility with Radix Scrophulariae and Angelicae Sinensis Radix.The parameters values of isochlorogenic acid C are the highest in SYD,but its statistical analysis have no significant difference.In Radix Scrophulariae,the parameters values of harpagide,angoroside C and harpagoside are all the highest after the herb compatibility with Lonicera Japonica and Angelicae Sinensis Radix,and lower in single Radix Scrophulariae group than in other DCGs.The absorption level of liquiritin in SYD is mediate,however its intestinal absorption parameters(Ka,Papp)values are the highset in Licorice.Part five:study on the compatibility of Simiao Yong’an Decoction based on prototype components into the blood by pharmacokinetics modelThe plasma sample content determination method for the seven components absorbed into blood of SYD using UPLC-MSn in negative ion mode technology is established.Linarin is selected as the internal standard.The quantitative ions are as follows:harpagide(m/z 183.11),chlorogenic acid(m/z 191.05),sweroside(m/z 125.07),loganin(m/z 227.12),liquiritin(m/z 255.13),angoroside C(m/z 175.05)and harpagoside(m/z 147.03).Each component shows good linear relationship in the individual range of 0.667-1558.660、0.807-1886.321、5.813-13580.340、1.690-3948.969、0.161-1003.754、0.121-2157.445、0.029-580.335 μg·mL-1.The matrix effects of low dose are from 80.590%to 111.896%,the RSD is from 2.420%to 9.730%.The matrix effects of middle dose are from 81.085%to 113.569%,the RSD is from 1.155%to 5.733%.The matrix effects of high dose are from 82.892%to 105.884%,the RSD is from 1.355%to 10.577%.The extraction recoveries of low dose are from 84.869%to 97.436%,the RSD is from 1.044%to 13.620%.The extraction recoveries of middle dose are from 82.900%to 99.485%,the RSD is from 3.017%to 6.776%.The extraction recoveries of high dose are from 86.785%to 98.187%,the RSD is from 0.810%to 8.572%.The established method has good method stability,short term stability,long term stability,freeze thaw stability with high precision and accuracy,which accordance with the detecting requirements of the biological sample.The method is used to detect the pharmacokinetic changes of the seven components in separated prescription experiments.Harpagide:The time to peak(Tmax)increases(P<0.01)markedly in SYD,and it is apparently higher(P<0.05)in three DCGs than that in two DCGs.There is no obvious difference of the area under the curve(AUC)among different DCGs,but its maximum value exists in single Radix Scrophulariae prescription.The value of mean residence time(MRT)in SYD is just inferior to that in the compatibility of Raix Scrophulariae and Licorice,and it’s higher(P<0.05)than other DCGs.There is no obvious difference of the t1/2 among different DCGs.Its plasma peak concentration(Cmax)value in SYD or single Radix Scrophulariae is higher(P<0.05)than that in other DCGs except the compatibility combination of Lonicera Japonica,Radix Scrophulariae and Angelicae Sinensis Radix.Moreover,the Cmax values in three DCGs are higher than that in two DCGs.Chlorogenic acid:The t1/2 in SYD is obviously increasing(P<0.01)compared to different DCGs.The peak value(P<0.01)of MRT exists in individual Lonicera Japonica,and three is not any obvious difference among other DCGs.The mean Tmax increases(P<0.05)when the four herbs compatibility with each other,and the value of this pharmacokinetic parameter is superior to any DCG except the compatibility of Lonicera Japonica,Angelicae Sinensis Radix and Licorice.The peak value(P<0.05 of Cmax exists in individual Lonicera Japonica,and the value of this pharmacokinetic parameter is superior to(P<0.05)any DCG except the compatibility of Lonicera Japonica and Radix Scrophulariae.Sweroside:The MRT and Tmax in SYD are both apparently higher than that in otherDCGs,however the statistical analysis does not show any significant difference.The t1/2 varies evidently in different DCGs,and the value of this pharmacokinetic parameter in single Lonicera Japonica and SYD is obviously decreasing(P<0.05)with the minimum value existing in SYD.Loganin:The AUC of loganin in SYD is evidently higher(P<0.05)than other DCGs.The MRT of loganin in SYD is evidently higher(P<0.01)than other DCGs.Its Tmax increases compared to other DCGs but the statistical analysis does not show any significant difference.The t1,2(P<0.01)and Cmax(P<0.05)in single Lonicera Japonica or SYD are obviously decreasing than that in other DCGs.Liquiritin:Its t1/2 prolongs(P<0.01)obviously than that in other DCGs.Among different compatibility combinations,the value of MRT decreases(P<0.05)evidently in single Licorice.The peak value of MRT exists in the combination of four herbs,while there is no significant difference via the comparison with other DCGs.The value of Tmax and Cmax of liquiritin are both lower in SYD than that in other DCGs.Angoroside C:The value of MRT in SYD is the highest,nevertheless there is no significant difference through comparing with other DCGs.TheAUC of angoroside C in SYD is significantly higher(P<0.05)than three DCGs.Among different compatibility combinations,the value of t1,2 decreases(P<0.05)evidently in single Radix Scrophulariae,while the value of t1/2 of angoroside C is lower in SYD than that in other DCGs.There are not any obvious differences of the value of Tmax among different DCGs,but the Cmax in SYD is increasing(P<0.05)significantly than that in other DCGs.Harpagoside:The t1/2 of harpagoside varies obviously in different compatibility combinations,the peak value exists in the compatibility of four herbs.The value of t1/2 is obviously higher(P<0.05)in three DCGs than that in two DCGs.The value of Cmax is obviously higher(P<0.05)in the compatibility of four herbs than that in other DCGs.The AUC of harpagoside in SYD is evidently higher(P<0.01)among different DCGs except for single Radix Scrophulariae.Innovative:The first time of identificating and source determinating of the serum chemical composition in Simiao Yong’an Decoction and its ingredients using LCMS-IT-TOF.Decoction compatibility research of the compositions absorbed into blood in Simiao Yong’an Decoction using UPLC-MSn for the first time.Intestinal absorption compatibility research of the compositions absorbed into blood in Simiao Yong’an Decoction using UPLC-MSn for the first time.Pharmacokinetics compatibility research of the compositions absorbed into blood inSimiao Yong’an Decoction using UPLC-MSn for the first time. |
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