ABSTRACT
OBJECTIVE:To establish HPLC fingerprint of stir-baked Manis pentadactyla.METHODS:HPLC method was conducted.The determination was performed on Capcell Pak Mg Ⅱ S5 C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution (gradient elution) at the flow rate of 0.8 mL/min.The detection wavelength was set at 275 nm,and column temperature was 30 ℃.The sample size was 10 μL.Using oxyphenylaminopropionic acid as reference,HPLC chromatograms of 11 batches of medicinal materials were determined.TCM Chromatogram Fingerprint Similarity Evaluation System (2004 A edition) was used for common peak identification and similarity evaluation.RESULTS:There were 23 common peaks in 11 batches of stir-baked M.pentadactyla,with similarity＞0.90.After validation,HPLC chromatograms of 11 batches of medicinal materials were in good agreement with control fingerprints.CONCLUSIONS:Established HPLC fingerprint can provide reference for the identification and quality evaluation of stir-baked M.pentadactyla.
ABSTRACT
OBJECTIVE:To establish HPLC fingerprint of stir-baked Manis pentadactyla.METHODS:HPLC method was conducted.The determination was performed on Capcell Pak Mg Ⅱ S5 C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution (gradient elution) at the flow rate of 0.8 mL/min.The detection wavelength was set at 275 nm,and column temperature was 30 ℃.The sample size was 10 μL.Using oxyphenylaminopropionic acid as reference,HPLC chromatograms of 11 batches of medicinal materials were determined.TCM Chromatogram Fingerprint Similarity Evaluation System (2004 A edition) was used for common peak identification and similarity evaluation.RESULTS:There were 23 common peaks in 11 batches of stir-baked M.pentadactyla,with similarity＞0.90.After validation,HPLC chromatograms of 11 batches of medicinal materials were in good agreement with control fingerprints.CONCLUSIONS:Established HPLC fingerprint can provide reference for the identification and quality evaluation of stir-baked M.pentadactyla.
ABSTRACT
The study was aimed to establish a stable, accurate site specific PCR identification system to identify Manis pentadactyla and its adulterants using DNA molecular identification. The genomic DNA was extracted from experimental samples using the DNA extraction kit. The Cytb and CO Ⅰ genes were amplified using PCR and sequenced bi-directionally. Obtained sequences were assembled using the BioEdit software. The neighbor-joining tree was constructed by MEGA 6.0. Specific identification primers were designed according to the specific allelets, and PCR reaction system was optimized. The results indicated that the Cytb and CO Ⅰ sequence both were able to be used to identify M. pentadactyla and its adulterants. With the specific primers CO Ⅰ-S10/A5, the M. pentadactyla could be amplified a 400 bp DNA band when the annealing temperature ranged from 55 to 60 ℃ and the amount of DNA template ranged from 3 to 100 ng within 35 PCR cycles. However, other adulterants displayed no relevant bands. So that primers CO Ⅰ- S10 / A5 can be used to identify the M. pentadactyla with the adulterants.