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OBJECTIVE To establisha simultaneous determination method of three main alkaloids in Plumula nelumbinis by high performance liquid chromatography(HPLC), including liensinine, isoliensinine and neferine, and optinize the ultrasonic-assisted extraction process of total alkaloids from Plumula nelumbinis by using response surface methodology based on Box-Behnken design. METHODS The alkaloids were separated on a RP-HPLC C18 monolithic column(4.6 mm×50 mm, Merck, Darmstadt, German) with a mobile phase consisting of methanol-water-triethylamine-acetic acid(70∶30∶0.2∶0.05), and the method was validated for linearity, sensitivity and extraction recovery. Based on the single-factor analysis, a mathematical model was constructed to analyze the effects of each factor of the ultrasonic-assisted extraction and their interactions on alkaloids yields of Plumula nelumbinis. The three independent variables were volume fraction of ethanol(A), solid-liquid ratio(B) and ultrasound extraction time(C), respectively. RESULTS The HPLC method used to separate three alkaloids within 5 min showed an excellent linear correlation (r2>0.999) in the range of 3.4-340 μg·mL-1(liensinine), 3.47-347 μg·mL-1(isoliensinine) and 3.47-347 μg·mL-1 (neferine). The limits of detection(LODs) of three alkaloids were 0.17, 0.69 and 0.69 μg·mL-1, and the limits of quantification were 0.34, 1.73 and 1.73 μg·mL-1, respectively. The intra-day and inter-day variations of RSD were less than 5%, and the extraction recovery rang from 97.06% to 104.12% with RSD ranging from 0.08% to 0.87%(n=6). The optimum extraction conditions were: solid-liquid ratio, 1∶30.3; volume fraction of ethanol, 69.8%; ultrasound extraction time, 29.0 min. The yield under the optimum conditions was found to be(14.82±0.42) mg·g-1, which was agreed closely with the predicted value of 14.70 mg·g-1. CONCLUSION The HPLC method used to determine the three main alkaloids in Plumula nelumbinis shows ideal characteristics of quickness, accuracy, high sensitivity and good repeatability.The optimum ultrasonic extraction technology has expressed excellent extraction ability of alkaloids in Plumula nelumbinis which indicates the RSM result is reasonable and effective.
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OBJECTIVE: To investigate the intestinal absorption behaviors of three main active components, liensinine, isoliensinine and neferine in Plumula Nelumbinis extracts in intestine of rats. METHODS: With phenol red as the indicator, in situ single pass intestinal perfusion (SPIP) model was used and the concentrations of the alkaloids in perfusion solution in different intestinal segments (duodenum, jejunum, ileum, and colon) were determined by HPLC. RESULTS: Liensinine, isoliensinine and neferine were absorbed in whole intestinal segments with saturation phenomena. The absorption rate constant (Ka) and/or effective permeability values (Peff) of the three alkaloids mostly had significant difference (P<0.05) at middle and high concentrations of perfusion solution vs those at low concentrations. Liensinine, isoliensinine and neferine possessed better absorption in ileum and colon of intestine. The absorption in duodenum and jejunum mostly displayed no significant difference (P≤0.05). In middle concentration group (117.3 μg·mL-1), neferine was absorbed best among the three alkaloids in colon; in other intestinal sections, the Kd and Peff of the three alkaloids were sequenced as follows: neferine ≤ liensinine ≤ isoliensinine. CONCLUSION: The transport mechanism of liensinine, isoliensinine and neferine in vivo may be active transport or facilitated diffusion.
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OBJECTIVE: To optimize the extraction conditions of plumula nelumbinis. METHODS: The semi - bionic extraction (SBE) conditions were optimized by homogeneous design while total alkaloids and dry extract were adopted as markers. Factors of extraction time, solute quantity and the pH values were evaluated for their effects on extraction process. RESULTS: Optimal parameters were as follows: plumula nelumbinis was extracted in water 8 times as much as it for 3 times; the pH values of the water for the 1st, 2nd and 3rd decoctions were 5.0,6.0 and 8.0, respectively; the boiling lasted 4 hours. CONCLUSION: The method prouides theoretical reference for the extraction plumula nelumbinis.
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OBJECTIVE:To optimize the extract technology for Plumula Nelumbinis.METHODS:Four methods:water extraction(WE),water extraction by precipitation with alcohol(WAE),semi-bionic extraction(SBE),and semi-bionic ex-traction by precipitation with alcohol(SBAE)were used to extract Plumula Nelumbinis and also compared in their effects by adopting the total content of alkaloid and the yield of dry extract as markers.The comprehensive values Y was calculated.RESULTS:Through the comprehensive evaluation of the two markers,the comprehensive values Y ranked in the order of SBE,WE,SBAE and WAE.CONCLUSION:SBE method is the optimal method to extract Plumula Nelumbinis.
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Objective To investigate the adsorption function of macroporous adsorption resins for the separation and purification of the effective components in Plumula Nelumbinis. Methods AKS-W, AB-8, and H214 resins were systematically studied for their adsorption capability, and the contents of the liensinine, isoliensinine, and neferine were determined by HPLC. Results AKS-W Resin had a preferable adsorptive and separateive effect for the effective components in Plumula Nelumbinis. The products containing 10.2% liensinine, 6.7% isoliensinine, and 11.9% neferine could be obtained. Conclusion Macro-porous-type and non-polar adsorption resin AKS-W is an ideal adsorbent for the extraction of the effective components in Plumula Nelumbinis.
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Objective To study the safety of Plumula nelumbinis microcapsules and its anti-arrhythmic effect.Methods Acute toxicity test in mice,arrhythmia mice models induced by chloroform and arrhythmia rats models induced by barium chloride(BaCl2)were adopted.Results The maximum tolerance dose for mice was 16.0 g/kg body weight.The microcapsule decreased the incidence of chloroform-induced ventricular fibrillation in mice significantly(P 0.05,compared with the blank control group).Conclusion The preliminary results show that Plumula nelumbinis microcapsule is safe,and exerts anti-arrhythmic effect against experimental arrhythmia.