Determination Of Pyrrolizidine Alkaloids And Their Nitrogen Oxides In Traditional Medicinal Materials Included In Chinese Pharmacopoeia

The medicinal materials in the pharmacopoeia were used to analyze the pyrrolizidine alkaloids(PAs)and their nitrogen oxides.This research can not only understand the distribution status of PAs in traditional medicinal materials,but also contribute to the formulation of relevant limit standards.It can also provide quantitative reference for the safe use and risk assessment of Chinese medicinal materials.The objective of this study was to establish a method for the analysis and determination of pyrrolizidine alkaloids in traditional Chinese medicinal materials.The method was applied to the qualitative and quantitative analysis of pyrrolizidine alkaloids and their nitrogen oxides in traditional medicinal materials from different sources.Therefore,we can obtain the preliminary results of the content and species of PAs in the medicinal materials recorded in pharmacopoeia.In addition,the results can provide experimental basis for the further study of these toxic components in Chinese medicinal materials.In this project,we selected the ultra-high performance liquid chromatography-tandem four-stage mass spectrometry(UPLC/MS/MS)and the purification method of solid phase extraction(SPE)to optimize and establish the analysis method of pyrrolizidine alkaloids in Chinese medicinal materials.This method has high sensitivity and can be used for the determination of trace amounts in Chinese herbal medicines.During the experiment,32 different PAs were used as reference to determine the species and content of pyrrolizidine alkaloids and their nitrogen oxides in the traditional Chinese medicinal materials included in the Chinese pharmacopoeia.The above method was applied to the PAs detection of different batches of lithospermum,flos farfarae,circium japonicum and sculellaria barbata samples to further explore the PAs content and distribution differences in different medicinal materials.Our research group and the other two research groups collected and tested 386 pharmacopoeia medicinal materials,271 of which were detected to contain PAs(content more than 0.1 ?g/kg).The approximate daily intakes of medicinal materials were estimated according to the dosage of pharmacopoeia,and there were 18 medicinal materials whose daily maximum intakes exceeded the EU standard of0.4248 ?g/day(adult body weight 60 kg).Our research group collected medicinal materials from different sources,and determined the content of PAs.In the determination,it was found that the average total amount of PAs in the flos farfarae was about 47.25 mg/kg.And the kinds of alkaloids are basically the same,but the content of each component is different.The most important PAs is senkirkine,which has significant hepatotoxicity.The mean total amount of PAs detected in the other three medicinal materials from different sources was significantly different,and the species distribution of the measured components was slightly different.The average amount of PAs in the lithospermum is 33.28 mg/kg,and echimidine-N-oxide is the main PAs component.The other components were unsaturated PAs with ring opening,and the toxicity was relatively weak.The contents of PAs in the circium japonicum and sculellaria barbata were relatively low,with an average total amount of about 0.0956 mg/kg and 0.0321 mg/kg.This is the first time to complete the determination of PAs components in these two herbs.The method used in this study is efficient,rapid and highly sensitive,which is suitable for the analysis and detection of pyrrolizidine alkaloids in Chinese medicinal materials.The test results of mixed samples of medicinal materials from different sources in the pharmacopoeia showed that most of the medicinal materials included in the pharmacopoeia generally contained PAs.Some commonly used medicinal materials were with high content,but without standard for the safe use at present.Therefore,more attention should be paid to this situation by relevant departments to carry out more extensive investigation and research to ensure the safe use of Chinese medicinal materials.It was found that there were some differences in the content and types of PAs in the medicinal materials from different sources after the detection of multiple batches of samples of four selected medicinal materials.Meanwhile,the distribution of different PAs compounds in medicinal materials was also different.Therefore,the investigation of PAs components in medicinal materials from different sources can ensure the accuracy of the results and the rationality of the standard formulation.