Study On The Fingerprint Of Mylabris

Object: Establishment of the GC-MS and the HPLC fingerprints of Mylabris for expliciting the differences on chemical composition of different habitats and species, then providing the evidence for its quality control.Method: Firstly, The conditions of solid phase micro extraction(SPME) and chromatographic of the GC-MS analysis of Mylabris were optimized through the resolution and the number of peaks. And then the methodologies of the GC-MS fingerprints were studied, which including: precision test, stability test and repeatability test. Finally, the 22 batches of Mylabris were analyzed, and then the fingerprint was created. The fingerprint similarity was calculated. At the same time, the quality of Mylabris was evaluated by fingerprinting and the measured results of the relative content of cantharidin.2、Firstly, The conditions of extraction and chromatographic of the HPLC analysis of Mylabris were optimized through the resolution and the number of peaks. And then the methodologies of the HPLC fingerprints were studied, which including: precision test, stability test and repeatability test. The fingerprint similarity was calculated. At the same time, the quality of Mylabris was evaluated by fingerprinting and the measured results of the relative content of cantharidin.3、The differences of chemical composition and quality between different habitats and species of Mylabris were analyzed by multivariate statistical analysis methods. In our research, the GC-MS and HPLC fingerprints and determination of cantharidin were Combined based on the similarity and the content of cantharidin of the two finger printings. These four parts were given the weight of 25%, and then classified the Mylabris to evaluate its quality according to the comprehensive score.Results: 1, 100 μm PDMS extraction head, 0.4 g sample, adsorption, equilibrium temperature of 80 ℃, equilibrium with 30 min, the adsorption with 15 min, resolution with 2 min were used for GC-MS SPME extraction. Agilent HP-5MS column(30 m×0.25mm×0.25 μm) was used for analysis. The flow rate was 0.5 m L·min-1, splitless with high purity He. The inlet temperature was 250 ℃, and the temperature program was that: the initial temperature was 50 ℃, then the temperature was raised to 250 ℃ with 3 ℃·min-1. The MS conditions were that: ionization mode was EI, EM voltage was 70 e V, quadrupole temperature was 150 ℃, ion source temperature was 230 ℃, mass scan range was 30 ~ 500 amu. The results of the precision, stability and reproducibility show that the relative standard deviation(RSD) of the relative retention time and relative peak area of total of 11 major peaks were less than 5%. Then the chromatographic fingerprint similarity evaluation system A version was used for evaluating it, the similarity of the spectrum between the sample and the control of the 22 batches Mylabris sample were greater than 0.906, five major peaks were calibrated. The results showed that the volatile components had no significant difference between Mylabris phalerata Pallas and Mylabris cichorii Linnaeus. However, the relative content of cantharidin is in large different from 38.86% to 64.12%. 2, Phenimenex Synergl 4u Polar-RP80 A column(250 mm×4.6 mm×5 μm) was used for analysis. Mobile phase was A- methanol, B-(1% acetic acid- water), gradient elution, injection volume was 20 μL, the flow rate was 1.0ml·min-1, column temperature was 30 ℃, detection wavelength was 254 nm, detection time was 30 min. T The results of the precision, stability and reproducibility show that the RSD of the relative retention time and relative peak area of total of 32 major peaks were less than 5%. Then the chromatographic fingerprint similarity evaluation system A version was used for evaluating it, the similarity of the spectrum between the sample and the control of the 22 batches Mylabris sample were greater than 0.781, only four batches of Mylabris were less than 0.900, the rest were greater than 0.900, while bound cantharidin contents in the extract of Mylabris were determined. However, the relative content of cantharidin is in large different from 0.12%-1.47%3, The comprehensive quality rating system of Mylabris preliminarily was established. By these four parts, the acquisition of Mylabris quality were divided into four grades, including one batch for first, 7 batches for second, 12 batches for third, and 2 batches for fourth.Consults: The GC-MS and HPLC fingerprints of Mylabris method were established in our experiment, and the methods have good reproducibility and precision, which will help us for better enhance the quality control of Mylabris. Finally, provide the evidence for its safe and rational clinical use of Mylabris.