| Study the chemical character of the active components alignment on Flos Trollii Chinensis and its recipe discovery the associativity of the crude drug and its recipe and the new methodology on synthetic evaluating quality.Adopt XRD, IR, LC and LC-MS to set up the finger identified standard of Flos Trollii Chinensis and its recipe, analysis the character on three orders of the active components alignment and set up the database on evaluating the active components alignment assisting PC. First, we investigated and identify the kind of the commodity Flos Trollii Chinensis and based on the main species, Trollii Chinensis Bunge, to develop research. Then, we alone use XRD, IR, HPLC and LC-MS to analysis the chemical character on Flos Trollii Chinensis and its recipe. At last, we can analysis the associatibity between the crude drug and the patent, know the applicability of every identified method and the reliability and the science of quality standard.Results: 1 The origin of the commodity Flos Trollii Chinensis is mainly came from Trollius chinensis Bunge that is the rated kind in Pharmacopeia. 2 XRDF indicated that Flos Trollii Chinensis in Neimeng has five special marks that are 8.17/14, 7.19/15, 5.42/21, 3.57/20 and 2.781/21, Flos Trollii Chinensis in Heilongjiang has seven special marks that are 10.05/15, 5.13/38, 4.94/19, 3.43/24, 3.11/18, 2.644/27 and 2.479/20 and the cultivated Flos Trollii Chinensis has not 3.22. So we can identify Flos Trollii Chinensis from the different producing place and the different ecological environment. Recipe Jinlianhua have three five special peaks (11.51/25,9.27/33,6.64/25,5.55/46, 5.13/38) that have not expressed in medicinal materials, so Px is 22.73 %, P is 77.27 %( this is the total rate of the recipe and the medicinal materials) by calculating. The special XRDF mark peaks of medicinal materials, Flos Trollii Chinensis has three special peaks (9.94/17, 8.17/14,5.42/21), and Px is 17.65 %. And its special interplanar spacing in the prescription is from 1.986 to 4.25. Flos Chrysanthemi Indici has one special peaks(8.38/35),and Px is 3.57 %. Its special interplanar spacing in patent is from 1.971 to 4.63, and the coincided interplanar spacing is 2.907 . Flos Lonicera Japonicae has two special peaks (4.95/15,4.50/16), Px is 9.09 %. And its special interplanar spacing in patent is from 2.055 to 4.95, the first coincided interplanar spacing is 3.93A and the second is 3.05A。The special XRDF mark peaks of Baicalin have nine peaks(10.34/62,8.59/23,6.49/10,6.05/34,5.23/44,3.86/28, 2.713/20, 1.881/10,1.752/10), and Px is 18.37 %. Its special interplanar spacing is from 2.016 to 4.44, and the coincided interplanar spacing is 2.016 .3 FT-IR indicated that it can identify Flos Trollii Chinensis of the different producing place and the second derivate spectrum can identify the different storage time.Recipe Jinlianhua have fifteen IR peaks and can be identified by the second derivate spectrum with 1 416 cm-1.4 HPLC fingerprint indicated that Flos Trollii Chinensis fingerprint owns 26 peaks (n=20) and assure 3 peaks by comparing the reference compounds. After oral the aqua extract of Flos Trollii Chinensis by rats, we can detect 6 compounds. There is no significant difference between the different places of production(RSD<1%). By the prolonged storage time, the contents of 2′′-O-β-L-galactopyranosyl orientin in Flos Trollii Chinensis present the depress trend and the contents of orientin present the heighten trend, but the together contents have no significant difference.The HPLC fingerprint of Recipe Jinlianhua owns twelve peaks by both and has two peaks only bodied in some samples. The former is more than 99% in the total area and the latter is less than 1%. The similarity scale is from 0.996 to 1.000.By analyzing the composition of the prescription, we can get the below conclusions. The character of the active constituents that the corresponding constituents of Flos Trollii Chinensis lies the retention time between 15min and 30min, which have seven peaks and mainly are flavanoid. The character of the active constituents that the corresponding constituents of Flos Chrysanthemi Indici lies the retention time between 5min and 10min and between 25min and 40min, which have seven peaks and mainly are phenolic acid and flavanoid. The character of the active constituents that the corresponding constituents of Flos Lonicerae Japonica lies the retention time between 5min and 10min and between 25min and 40min, which have nine peaks.Analyzing the classification of the compounds the flavanoid lies the retention time from 15min to 50min and the phenolic acid from 5min to 15min. Through analyzing the compound, we find that 2′′-O-β-L-galactopyranosyl orientin (tR=16.333min), orientin(tR=17.746min) and another peak(tR=52.18min) came from Flos Trollii Chinensis. The peak area ratio of 2′′-O-β-L-galactopyranosyl orientin and the peak(tR=52.18min) is 1/3 and they are the special active peaks from Flos Trollii Chinensis in the recipe. Chlorogenic acid that its retention time is 7.901min and caffeic acid (tR=10.384min) came from Flos Chrysanthemi Indici and Flos Lonicera Japonica. The retention time of Baicalin is 42.211 min.5 LC-MS fingerprint indicated that 2′′-O-β-L-galactopyranosyl orientin (tR=11.927min, M+ is 609), orientin(tR=13.949min,[M-H]- is 447) and vitexin (tR=16.589min, [M-H]- is 431) are came from Flos Trollii Chinensis. And we inferred eight peaks by analyzing the mass data that they are Orientin-2′′-O-β-D-pyranxyloside (tR=12.935min, [M-H]- is 579), 2′′-O-β-L-galactopyranosyl vitexin (tR=13.791min, [M-H]- is 593), 2′′-O-(3′′′,4′′′dimethoxylbenzoyl) vitexin (tR=14.723min, [M-H]- is 595), isoswertisin (tR=26.382min, [M-H]- is 447), 2′′-O-(2′′′methylbutyryl)orientin (tR=30.729min, [M-H]- is 531), quercetin (tR=34.332min, [M-H]- is 301), 2′′-O-(2′′′methylbutyryl) -vitexin (tR=35.315min, [M-H]- is 515) and 2′′-O-(2′′′methylbutyryl) isoswertisin (tR=43.583min, [M-H]- is 529)。We assured five compounds in LC-MS of Flos Chrysanthemi Indici that are acacetin(tR=57.54min, [M-H]- is 283), lianarin(tR=31.50min, [M-H]- is 591), luteolin (tR=33.21min, [M-H]- is 285), caffeic acid(tR=10.16 min, [M-H]- is 179) and cholorogenic acid(tR=7.41min, [M-H]- is 393) and inferred three compounds by MS data that are isoquercitrin (tR=18.59min, [M-H]- is 463), cosmosiin (tR=22.67min, [M-H]- is 431) and luteolin-7-O-glu(tR=18.16, [M-H]- is 447). By analyzing the mass data, we assured five compounds of Flos Lonicera Japonica that are hyperin(tR=17.98min, [M-H]- is 463), luteolin(tR=33.08min, [M-H]- is 285), caffeic acid(tR= 9.95min, [M-H]- is 179) and cholorogenic acid (tR=7.39min, [M-H]- is 393) and inferred luteolin-7-O-glu (tR=18.08min, [M-H]- is 447).Last we assured eight compounds that are cholorogenic acid(tR=7.47min), cafeeic acid(tR=10.027min), 2′′-O-β-L-galactopyranosylorientin(tR=12.38min), orientin (tR=13.763min), vitexin(tR=16.431min), hyperin(tR=18.991mi), baicalin(tR=28.83min), acacetin(tR=57.77min).The fingerprint of Flos Trollii Chinensis and its recipe can present the corresponding specificity. XRDF can identify the recipe and its composition that is crude drug, and can be used to identify the different producing place and the different ecological environment of the same species. IR fingerprint can identify Flos Trollii Chinensis came from the different producing place and the second derivate IR spectrum can be used to identify the storage time of Flos Trollii Chinensis and the quality of Jinlianhua Recipe. XRD and IR can evaluate the quality of Flos Trollii Chinensis and Jinlianhua Recipe by analyzing the whole compositions under that we have not full standards. HPLC fingerprint can evaluate the quality of Flos Trollii Chinensis and its recipe, trail the change rule of the active components alignment and can set the quality specification based on the active components alignment. LC-MS fingerprint can connect LC with MS and obtain more chemical character for evaluating the quality on Flos Trollii Chinensis and its recipe.On evaluating the quality, XRD can evaluate in the composition from the crude drug, IR in the crude drug and the compound, HPLC in the crude drug, the classification of chemical compounds and the compound and LC-MS in the compound.The new ideas of the paper as follows:1 We first set up four kinds of fingerprint character to evaluating quality of Flos Trollii Chinensis and its recipe.2 Propose the three orders of the active constituents on Chinese materia medica Chemistry what are the crude drug, the kinds of chemical compounds and the compound and set up the positioning mode of the active components. 3 Propose the associativity of the crude drug and the recipe, the quality criteria and the active constituents that used in evaluating the quality of Chinese materia medica.4 In the studying the character of XRDF identification, we raise that establishing the quality synthetic standard is base of the owned by both special marks in the medicinal materials of the patent and the coincidence peak data. By studying the IR fingerprint, we realize that identify the different producing place by one dimension IR and the different storage time by the second derivative spectrum. On HPLC fingerprint, we proposed that can analysis the chemical character in the crude drug, the kinds of chemical compounds and the compound. We raise that can monitor the separation of the active constituents and the technology of Jinlianhua Recipe and evaluate the quality by calculating the peak area. 5 Obtain 2 invented patents on Jinlianhua Recipe(one of the main inventors) and the patent is open. |
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