Evaluation Of The Quality And Safety Of Menthae Haplocalycis Based On Active Ingredient Content,Fingerprints And Pesticide

Objective:To establish a method for determining the content of menthol and the fingerprint of volatile components in Medicinal Mint by gas chromatography,and to establish a method for determining the content of four main non-volatile components and the fingerprint of non-volatile components in Medicinal Mint by high-performance liquid chromatography,explore the difference between mint and its confusions in terms of non-volatile ingredients,in order to provide a reasonable basis for the overall quality evaluation of the two major active ingredients of mint,and lay a certain theoretical foundation for the more reasonable and effective evaluation and control of mint quality in the future;To investigate and understande the pesticide residue status of nationwide mints,in order to realize the pesticide residue detection and a more comprehensive and safe quality evaluation of mints.Methods:(1)The GC method was used to establish content determination method of the menthol and the GC fingerprint of volatile components: the inl et temperature was set at 200 °C,the detector temperature was set at 300 °C,temperature was programmed,the split ratio was 10: 1,and each common pe ak was confirmed by the reference substance.(2)The DAD-HPLC technology wa s used to establish a method for determining the content of four main compon ents: linarin,caffeic acid,hesperidin,and rosmarinic acid in mint,and the cont ent was compared with spearmint and Hebei peppermint,which are easily mixe d.To establish HPLC fingerprints of non-volatile components,and combine sim ilarity evaluation and chemometric methods for analysis.The mobile phase was acetonitrile-0.2% phosphoric acid,and the gradient elution procedure was adop ted,the column temperature was controlled at 30 °C,the detection wavelength was 245 or 325 nm.(3)Based on the previous liquid chromatography,through o bserving the chromatograms of the three medicinal materials,It is found that t he three peaks that retention times respectively is 18.2,36.0,and 41.4 in the f igure are owned by mint,while Hebei mint and spearmint are almost not.It c ould be a distinctive component of 3 kinds of medicinal materials,so it is inte nded to confirm and determine these components by LC-MS.(4)GC-MS technology was used to establish the detection method of 28 pesticide residues in min t,and test 111 batches of mints.Results:(1)The content determination results of GC technology showed th at menthol content in mint decoction pieces should not be less than 0.11% on a dry basis;among the 192 batches tested,the pass rate was 66.1%,and the f ail rate was 33.9%;statistics on the fraction defective of samples from various origins show that the fraction defective in Jiangsu is the lowest,and the fracti on defective in Hebei and Sichuan is higher.The GC fingerprint results showe d that the limit similarity of the fingerprints should not be lower than 0.80;a mong the 192 batches of samples,the pass rate was 64.6% and the fail rate w as 35.4%,and it is consistent with the results of menthol research.(2)HPLC co ntent determination results showed that the content of caffeic acid,hesperidin,r osemary acid,and linarin in mint respectively is among 0.0080~0.1018 mg/g,0.1498~5.0057 mg/g,0.0103~3.3728 mg/g,0.0493~13.6978 mg/g;compared with the av erage content of the four components of spearmint and Hebei mint samples,it was found that the caffeic acid content of mint was basically the same as tha t of spearmint and Hebei mint,while the contents of hesperidin,rosemary acid,and linarin were higher than Spearmint and Hebei Mint.HPLC fingerprint an alysis showed that there were 14 common peaks in mint,the similarity evaluat ion software,cluster analysis,and partial least squares discriminant analysis we re used to evaluate mint,spearmint,and Hebei mint,the results were consistent.(3)Confirmed by using liquid chromatography-mass spectrometry,the three co mpounds were inferred to be succinic acid(3,4-dihydroxybenzylidene),orthopuri c acid,and locustin,and they all were found in mint,Hebei mint,and spearmi nt,indicating that under the current liquid phase conditions,the distinguishing components of the three cannot be found.(4)Pesticide residue test results sho wed that in 111 batches of mint samples,phorate was detected in 1 batch of s amples,p,p'-didiyi was detected in 1 batch of samples,and metalaxyl was det ected in 3 batches of samples,the remaining pesticides were not detected.Conclusion: The established method for the determination of menthol and 4non-volatile components provides a basis for improving the quality control of activeingredients of mint;The established fingerprint method of volatile and non-volatile components shows that the method can effectively determine the authenticity of mint,comprehensively control its quality,and provide a basis for the overall quality evaluation and control of mint.QuECHERS-gas chromatography-tandem mass spectrometry was used to detect 28 pesticide residues.This method is fast and simple,and can provide a basis for the safety evaluation of mint.It provides research ideas for revising and improving the quality standards of peppermint herbs and decoction pieces in the future,and assists in the research and development of quality and safety standards of traditional Chinese medicine.