RESUMO
Objective: To establish a rapid on-site method for identifying Chinese medical material recombinase-mediated amplification (RAA) technology for the use of identifying medicinal Bubali Cornu from yak horn. Method: Based on the differences of mitochondrial genome sequences between Bubali Cornu and adulterants,the specific RAA primer (SNJ-1.F,SNJ-1.R) and fluorescence probe SNJ-1.probe were designed by variation sites. Alkaline lysis method was used to extract DNA from milled samples,and optimize RAA reaction system. The incubation was made at 37℃ for 15-20 min, the reaction results were monitored through gel electrophoresis and a mobile fluorescence amplification instrument. The RAA identification result was compared with COI DNA sequencing. Result: After incubation at 37℃ for 20 min,about 140 bp of bright and simple bands was amplified from DNA templates of Bubalus bubalis,whereas Bos mutus were negative. By the Real-time fluorescent RAA identify method,all reactions in DNAs from Bubali Cornu samples were amplified from 6.21 to 8.37 min,whereas DNAs from yak samples were amplified after 10.08 min,COI sequencing results conformed to the Real-time fluorescent RAA identification. Conclusion: Specific RAA could rapidly identify Bubali Cornu in 20 minutes, and thus be applied in medical material and its products. This method is simple,rapid and sophisticated instrument-free,with promises in on-site identification of traditional Chinese medicine.
RESUMO
A fingerprint method for quality assessment of Fritillaria thunbergii was developed by rapid resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (RRLC-Q-TOF-MS). The separation was performed using Agilent Eclipse Plus C18 column (2.1 mm x 100 mm, 1.8 microm) by gradient elution with acetonitrile and 0.1% formic acid aqueous solution (containing 10 mmol x L(-1) ammonium formate) as the mobile phase. Q-TOF-MS was used to obtain the accurate mass for precursor and product ions. Under this chromatographic and MS condition, 12 batches of F. thunbergii and its adulterants (F. hupehensis and F. pallidiflora) were analyzed by RRLC-Q-TOF-MS. Fifteen steroidal alkaloids were identified from F. thunbergii, F. hupehensis and F. pallidiflora and nine were assigned as the common characteristic peaks for F. thunbergii. The RRLC-Q-TOF-MS fingerprint of F. thunbergii was different significantly with those of F. hupehensis and F. pallidiflora. The quality of 12 batches of F. thunbergii samples were finally evaluated by hierarchical clustering analysis (HCA) and principle component analysis (PCA). This convenient and high specific method could be used to identify and evaluate the quality of the F. thunbergii.
Assuntos
Alcaloides , Química , China , Cromatografia Líquida de Alta Pressão , Métodos , Medicamentos de Ervas Chinesas , Química , Fritillaria , Química , Classificação , Controle de Qualidade , Espectrometria de Massas por Ionização por Electrospray , Métodos , Espectrometria de Massas em Tandem , MétodosRESUMO
<p><b>OBJECTIVE</b>To identify triacylglycerols in coix oil.</p><p><b>METHOD</b>High performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry was used for identification. The experiment was operated under the conditions: spray voltage at 3 000 V, capillary temperature at 250 degrees C, APCI vaporizer temperature at 400 degrees C, and corona current of 4 microA. Sheath gas pressure (high purity liquid nitrogen) was 35 kPa. Mass spectra were obtained over the m/e range of 300 to 900 amu, scan duration of 1s and Q1 peak width at 0.7. The stationary phase was Zorbax Extend C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase: dichloromethane-acetonitrile (35:65), flow rate: 1 mL x min(-1); column temperature: 25 degrees C.</p><p><b>RESULT</b>12 triacylglycerols were identified by HPLC-MS method.</p><p><b>CONCLUSION</b>The result can be used to identify the components in a fingerprint chromatogram of coix oil and its related injection product.</p>