Study On Separation And Analysis Of Alkaloids In Macleaya Cordata

Objective:The first part of this article is conducted to establish a handy,convenient,speedy and effective capillary electrophoresis method for the simultaneous separation and analysis of sanguinarine and dihydrosanguinarine,and apply it into the leaves and roots of Macleaya cordata(Willd.)R.Brown.Additionally,the proposed method provides a method basis for the analysis of alkaloids in Macleaya Cordata.The second part of this paper adopts direct ultraviolet spectrophotometry and acid dye colorimetry to determine the total alkaloids in the leaves and roots of Macleaya cordata,respectively.The method is simple and rapid,and the results are accurate and reliable.Moreover,it can be used to control the quality of the Macleaya Cordata and its preparations.Methods:1.The experiments provided a simple approach for the optimization of CE separations,a P/ACE MDQ CE system was carried out,which equipped with a diode array detector(DAD).The influence of the type and concentration of the buffer solution,the pH,separation voltage and temperature on the separation of Macleaya Cordata were investigated.The optimum conditions of capillary electrophoresis were as follows: a fused stretchy uncoated silica capillary column(40.2 cm × 75 μm id,with an effective length of 30.0 cm)was adopted,the concentration of 5.0 mmol/L sodium dihydrogen phosphate solution(pH 4.20)was employed as the buffer solution,the separation voltage was 20 kV.Samples were injected into capillary column under the pressure of0.5 psi for the duration of 5.0 s.The study picked 270 nm as the detection wavelength,and at a temperature of 25℃.2.A UV-1200 ultraviolet spectrophotometry was employed.And the type,pH and dosage of medium buffer solution and the amount of acid dye bromocresol green solution were studied.Finally,the optimum colorimetric conditions for acid dyes were determined,which included pH 4.80 disodium hydrogen phosphate 5.0 mL,2.5 mL bromocresol green solution,organic reagent was 10 mL chloroform,the determination wavelength at 410 nm,and the temperature was 25℃.Results:1.The first part of the experimental results showed that under the optimal electrophoretic conditions,sanguinarine and dihydrosanguinarine achieved baseline separation within less than 4 min,the linear relationships of the two alkaloids were good,and the correlation coefficient was in the range of 0.9986-0.9993.The values of RSD of intraday precision were in the range of 0.9% to 1.5%,while the results of RSD of interday precision ranged from 0.4% to 2.5%.The average recoveries of the two alkaloids in the leaves and roots of Macleaya Cordata were from 99.6% to 102.8%.2.From the experimental results of the second part,we could find that:(1)When direct ultraviolet spectrophotometry was employed,a good linear relationship between concentration and absorbance was obtained as the concentration of sanguinarine in the range of 1.0 to 6.0 μg/mL,and the linear equation was y = 0.1045x-0.0216(r=0.9999,n=6).The average recoveries of the samples were 103.0%,102.7%,respectively.(2)When acid dye colorimetry was used,under the optimal medium conditions,a good linear relationship in the range of 3.0 to 8.0 μg/mL of sanguinarine was received,and the regression equation was y = 0.1123 x + 0.0104(r=0.9996,n=6).The average recoveries were 104.1%,103.7%,respectively.Conclusions:In this paper,a simple and efficient capillary electrophoresis method was developed to simultaneous separate and determine the two alkaloids in the leaves and roots of Macleaya cordata.This method provides a reference standard for the separation and analysis of active components in Macleaya cordata and its preparations.In this paper,the content determination of total alkaloids in the leaves and roots of Macleaya cordata was established by direct UV spectrophotometry and acid dye colorimetric method.It can be concluded that the above proposed analytical methods are easy to achieve and operate.Besides,both analytical approaches provide a scientific standard for the quality control of the Macleaya Cordata and make it better developed and utilized in the future time.