RESUMO
A high performance liquid chromatography( HPLC) method was established for the fast,and precise determination of ten nucleosides in Fritillariae Cirrhosae Bulbus and its counterfeits. Then multivariate statistical analyses,such as clustering analysis,principal component analysis( PCA),and Fisher' s linear discriminant analysis( LDA),were conducted to establish a discriminant function model for an integrated analysis. The results indicated that data acquisition time of a single sample was shortened within 16 min by the HPLC method. In the range of 5-1 000 mg·kg~(-1),the mass concentrations of all nucleosides exhibited good linear relationships with the corresponding peak areas( R2> 0. 999). The spiked recoveries were in the range of 93. 83%-108. 9% with RSDs of0. 12%-1. 3%( n = 5). The limit of quantitation( LOQ) was 0. 98-4. 13 mg·kg~(-1). As revealed by the clustering analysis,Fritillariae Cirrhosae Bulbus and the counterfeits could be discriminated into two clusters based on the content of nucleosides. Fisher's LDA could achieve this discrimination,while PCA dimension reduction failed. The accuracy of the discriminant function model established on the screened characteristic indicators reached 97. 5%. The present study proposed a new identification method of Fritillariae Cirrhosae Bulbus with one-dimensional indicators,which is simple,accurate,and reliable. It can provide a scientific basis for further optimizing the identification techniques for Fritillariae Cirrhosae Bulbus and inspiration for quality control strategy development of Chinese medicinal materials.
Assuntos
Medicamentos de Ervas Chinesas , Fritillaria , Cromatografia Líquida de Alta Pressão , Nucleosídeos , Raízes de PlantasRESUMO
The Chinese cordyceps, a complex of the fungus Ophiocordyceps sinensis and its species-specific host insects, is also called "DongChongXiaCao" in Chinese. Habitat degradation in recent decades and excessive harvesting by humans has intensified its scarcity and increased the prices of natural populations. Some counterfeits are traded as natural Chinese cordyceps for profit, causing confusion in the marketplace. To promote the safe use of Chinese cordyceps and related products, a duplex PCR method for specifically identifying raw Chinese cordyceps and its primary products was successfully established. Chinese cordyceps could be precisely identified by detecting an internal transcribed spacer amplicon from O. sinensis and a cytochrome oxidase c subunit 1 amplicon from the host species, at a limit of detection as low as 32 pg. Eleven commercial samples were purchased and successfully tested to further verify that the developed duplex PCR method could be reliably used to identify Chinese cordyceps. It provides a new simple way to discern true commercial Chinese cordyceps from counterfeits in the marketplace. This is an important step toward achieving an authentication method for this Chinese medicine. The methodology and the developmental strategy can be used to authenticate other traditional Chinese medicinal materials.
Assuntos
Cordyceps/genética , Medicamentos Falsificados/análise , Medicamentos de Ervas Chinesas/análise , Fraude/prevenção & controle , Reação em Cadeia da Polimerase , Animais , Cordyceps/química , Medicamentos Falsificados/química , Medicamentos Falsificados/economia , DNA Fúngico/isolamento & purificação , Medicamentos de Ervas Chinesas/economia , Medicamentos de Ervas Chinesas/normas , Complexo IV da Cadeia de Transporte de Elétrons/genética , Fraude/economia , Genes Fúngicos/genética , Genes de Insetos/genética , Proteínas de Insetos/genética , Insetos/genética , Insetos/microbiologiaRESUMO
An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/ MS) method was developed for the determination of citrus red 2 (CR2) in navel orange. The sample was extracted with acetonitrile, cleaned-up by an NH2 solid phase extraction cartridge. Then, the analyte was separated on a Waters Acquity UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 microm) in gradient elution with the mobile phases of water and acetonitrile. The separated compounds were detected with a Waters Xevo TQ MS tandem quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) mode via positive electrospray ionization (ESI+). The linearity of the method was good (r2 = 0.996 5) over the concentration range from 0.1 to 100 microg/L for CR2. The limit of detection (LOD) was 0.05 microg/kg, and the limit of quantification (LOQ) was 0.1 microg/kg. The average recoveries of CR2 at the three levels of 1.0, 10.0 and 100.0 microg/kg were between 84.2% and 94.0% with the relative standard deviations (RSDs) between 2.86% and 10.15%. Finally, a total of 18 navel orange samples randomly collected from different markets were detected by the proposed method, and two samples contained CR2. The sensitivity, accuracy, and precision of this method can meet the requirements of the CR2 analysis.