Research On Thin Layer Chromatography Of Nelumbinis Stamen,Leonuri Fructus And Leonuri Herba

In the Chinese Pharmacopoeia(2015 edition),the standard for Nelumbinis Stamen was recorded including the source,character,powder identification,property and flavor,channel tropism,function and main treatment,usage and dosage,and storage.The TLC identification was not recorded in the Ch P.Stachydrine hydrochloride was used as a chemical marker in the TLC identification of Leonuri Fructus and Leonuri Herba in the Ch P.Actually,the reaction between stachydrine and derivatization reagerd,dilute potassium bismuth iodide or dilute potassium bismuth iodide-ferric trichloride(10:1,V/V)was negative which makes the alkaloid spot undetected.To establish the TLC identification of Nelumbinis Stamen,the constituents were performed.To improve the reaction sensitivity of TLC derivatization of Leonuri Fructus and Leonuri Herba,the composition of reagerd was optimized.1 Study on the chemical constituent of Nelumbinis StamenTo establish the TLC identification of Nelumbinis Stamen by specific chemical marker,the constituents of Nelumbinis Stamen was carried on.In the literature report,the cycloaltun triterpenoids with anti-inflammatory activity activity were main components of the Nelumbinis Stamen.The ethyl acetate fraction(rich of the cycloaltun triterpenoids)was isolated and purified.A total of 16 compounds were identified from this fraction,including 2 new compounds(Nelmnucifergenin A;Nelmnucifergenin A)and 10 compounds(28-Hydroxyolean-12-en-3-one;Cycloarta-23,25-dien-3-one;25-Hydroxycycloart-23-en-3-one;Cycloart-25-ene-3?,24-diol;Macrostachyoside B;5,8-Epidioxyergosta-6,22-dien-3-ol;6-Hydroxy-stigmasta-4-en-3-one;N-Octacosan-7?-ol;N-Octacosan-7?-ol)isolated from Nelumbo the Genus for the first time.The new component Nelmnuciferoside A was selected as the marker of Nelumbinis Stamen based on which TLC identification of Nelumbinis Stamen.2 Study on alkaloid derivatization reagent of Leonuri Fructus and Leonuri HerbaIn order to solve the derivatization problem of stachydrine hydrochloride in Leonuri Fructus and Leonuri Herba,the alkaloid derivatization reagent was improved in this paper.After formulated the derivatization agent,the content of bismuthsubnitrate,potassium iodide and iodine in derivatization reagent was optimized by using response surface method.The composition of derivatization reagent was 0.82 g bismuth subnitrate,potassium iodide 11.06 g and iodine 0.76 g.Finally,the optimal volume of dilute phosphoric acid solution was 90 m L,and the developed method was validated.The limit of detetion for stachydrine hydrochloride and choline chloride were 2.0 ?g(1.93%)and 1.0 ?g(2.34%).The %RSD of inter-plate,intra-day and intra-day precision was less than 5.0%,and the %RSD of stability 15-90 min was4.79%.The method has high sensitivity and good stability.The derivatization agent was used for the TLC identification of Leonuri Fructus and Leonuri Herba.And in addition to stachydrine hydrochloride,the differential constituents and same constituents in these two herbs were detected.And TLC-EFISI-MS was used to characterize the different constituents and same constituents in Leonuri Fructus and Leonuri Herba.In addition,derivatization agents was expanded to TLC identification of Trigonellae Semen and Kanggongyan capsule.3 Studies on the TLC Identification of Nelumbinis Stamen,Leonuri Fructus and Leonuri HerbaTo establish TLC identification of Nelumbinis Stamen,a systematic methodological investigation was carried out on the thin layer chromatography of Nelumbinis Stamen.The optimum thin layer chromatography conditions were screened out,that is,the extraction solvent was methanol;the extraction method was ultrasonic treatment of 30 minutes;the reference solution volume was 2 ?L,the reference drug solution and test solution volume was 8 ?L,respectively;the thin plate was selected as the Merck plate,and the developing solvent was petroleum ether-ethyl acetate-formic acid(98%)(7:3:0.6,V/V/V).The derivatization agents was selected as the best solution of 10% sulfuric acid ethanol solution.Temperature was chosen to room temperature;The humidity range was between 42% and 72%.The new component Nelmnuciferoside A and reference drug were selected as the marker of Nelumbinis Stamen based on which TLC identification of Nelumbinis Stamen.By optimizing condition of the TLC of Leonuri Fructus and Leonuri Herba,the TLC identification of Leonuri Fructus and Leonuri Herba was modified by improved derivatization agent.