The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.

OBJECTIVETo establish a quantitative method with precolumn derivatization for determining the contents of six common amino acids in Banlangen Keli by UPLC.

METHODUsing 6-acetamido-4-hydroxy-2-methyl quinoline as the derivating agent, we determined the contents of arginine, threonine , alanine, gamma aminobutyric acid, proline, and valine. The UPLC analysis was performed on a Waters AccQ Tag TM Ultra C18 column (2.1 mm x 100 mm, 5 microm) with mobile phase AccQ Tag Ultra Eluent A and AccQ Tag Ultra Eluent B gradient elution at a flow rate of 0.7 mL x min(-1). The column temperature was 55 degrees C and detection wavelength was 260 nm.

RESULTThe linear ranges of arginine, thremine, alanine, gamma aminobutyric acid, proline, and valine were 4. 15549.86 microg (r = 0.999 9), 0.595-5.95 microg (r = 0.999 8), 0.445-4.45 microg (r = 0. 999 9), 0.515-5. 15 pg (r = 0.999 9), 8.858-106.3 microg (r = 0.999 9) , 0.585-5. 85 microg (r = 0.999 8). Their average recoveries were 100.6%, 98.35%, 100.2%, 98.44%, 98.34%, 98.18% with RSD 1.8%,1.9%, 2.0%, 2.4%, 1.5% and 2.0%, respectively (n = 6). The contents of amino acids were different in samples from five productive enterprises.

CONCLUSIONThe method is efficient, good reproducible, sensitive, and accurate.

Amino Acids ; analysis ; Chromatography, Liquid ; Drugs, Chinese Herbal ; chemistry ; Reproducibility of Results