Determination Of Scopolamine Alkaloids In Biological Samples By Ultra Performance Liquid Chromatography-mass Spectrometry

Mandala,Scopolamine,belladonna and other solanaceae plants are common poisonous plants in China,and they are also common in traditional Chinese medicine and ornamental plants.The main toxic component of these plants is scopolamine alkaloid.At present,due to misuse and illegal medical practice of above plants and other causes,scopolamine alkaloid poisoning cases have occurred.It is necessary to establish a rapid,accurate and effective method for the detection of scopolamine alkaloids in biological samples.In this paper,we studied three kinds of scopolamine alkaloids in Scarletidae: scopolamine,anisodamine and atropine.The experimental process is as follows.1.The chromatographic and mass spectrometric methods for the determination of scopolamine,anisodamine and atropine were established.2.The methods for extraction and purification of three alkaloids in biological samples were established,including precipitation protein method,liquid-liquid extraction method,solid-phase extraction method.3.The effectiveness of established methods was validated by which the scopolamine alkaloids were detected in practical cases.Experimental results.1.The column is Agilent Eclipse Plus ? C18(2.1 x 100 mm,1.8?m).The organic phase of the mobile phase is acetonitrile.The aqueous phase was a 0.1%formic acid aqueous solution.The gradient elution procedure was used.Quantitative ion pairs of scopolamine,atropine and anisodamine were 304.1> 138.0,290.1> 124.0,and 306.1> 140.0respectively.And qualitative ion pairs were 304.1> 138.0,304.1> 156.1(scopolamine);290.1>124.0,290.1> 93.0(atropine);and 306.1> 140.0,306.1> 90.9(anisodamine).Determine the minimum detection limit with S/N?3.Determine the minimum limit with S/N?10.The minimum detection limits for scopolamine,atropine and anisodamine were 0.02 ng / mL,0.02 ng / mL,0.04 ng / mL,respectively.The minimum limits of quantitation were 0.05 ng / mL,0.05 ng / mL,0.1 ng / mL,respectively.The method is linear in the range of 1~1000 ng / mL.R2 is above 0.99.2.The absolute recoveries of scopolamine,atropine and anisodamine were87.79% ~104.23%,85.96% ~106.76% and 86.14% ~96.39% respectively using the precipitated protein method,and the relative standard deviations of the recoveries were less than 7.06%.The absolute recoveries of scopolamine,atropine and anisodamine were 61.61% ~82.96%,68.36%~95.59% and 57.76% ~70.98% respectively,using liquid-liquid extraction method,and the relative standard deviations of the recoveries were below 7.70%.The absolute recoveries of scopolamine,atropine and anisodamine were 85.48% ~118.95%,87.24% ~102.63% and80.09% ~119.31% respectively,using solid phase extraction method,and the relative standard deviations of the recoveries were below 8.77%.The three pretreatment methods were linear in the range of 1~100 ng / mL for scopolamine,atropine and anisodamine,and RSDs of intra-and inter-day were below 8.62%,which would meet the detection requirements of practical cases.