The Technology Study On The Determination Of Chemical Components Illegally Added In Traditional Chinese Medicine

Hypertension and Diabetes are common disease on common clinical research presently, and the incidence has increased year by year. Some manufacturers illegally added chemical components in Traditional Chinese Medicine formulations, which intends to get more profits. This behavior brings large harm to people’s health. Methods of thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) were developed to determine the chemical components illegally added in TCM formulations for Hypertension and Anti-diabetes. The method developed was easy and effective, it can be supportive to the detection of these two kinds of medicines.The TLC and HPLC methods were developed to determine the Glipizide, gliclazide and glibenclamide possibly added in TCM formulations. The TLC method was used preliminary, silica gel GF254 was used as solid phase, and the elution system was Ethyl acetate-Cyclohexane-Ethanol-glacial acetic acid (8:15:2:1). TLC layers were observed under ultraviolet lamp with the wavelength of 254 nm. The method developed was fast and easily operated. The HPLC method was developed, chromatography separation was performed on Diamonsil C18 column (4.6 mm×200 mm,5μm)by using a mixture of methanol-0.5% glacial acetic acid(65:35) delivered at a flow rate of 1 mL-min-1 and detected at 230 nm. The column temperature was set at 30℃. The linear range were 4.08-81.6μg-mL-1(r= 0.9997)、4.04-80.8μg·mL-1(r= 0.9997)、4.12-82.4μg-mL-1(r= 0.9993), and the reproduction(n=9) were 97.7%,99.4% and 102.5% respectively. The LOD were 10.8 ng,9.6 ng,12.8 ng. Using this method, we detected 9 kinds of TCM formulations, one was detected with glibenclamide compound. We determined it under HPLC-MS, comparing with the Reference Materials, we proved that this TCM durg was illegally added with glibenclamide.The TLC and HPLC methods were developed to determine the Amlodipine Besylate, Nicardipine hydrochloride, Nifedipine, Nimodipine, Nisoldipine and Felodipine possibly added in TCM formulations. The eluent system, Ethyl acetate-Cyclohexane-Methanol-glacial acetic acid(8:8:1:0.1) was used to Amlodipine Besylate, Nicardipine hydrochloride and Nisoldipine, Ethyl acetate-Cyclohexane (4:5) was used to Nifedipine, Nimodipine, Felodipine. TLC layers were observed under ultraviolet lamp with the wavelength of 254 nm. The HPLC method was developed to determine 6 chemical drugs simultaneously. The column was Diamonsil C18 column, with the mobile phase of acetonitrile-0.03 mol/L Potassium dihydrogen phosphate (pH=3.0) with gradient elution, in 10 minutes acetonitrile from 65% to 55%,10 to 40 minutes, acetonitrile change to 25%. The column temperature was set at 35℃, the detection wavelength was 235 nm, the flow rate was 1 mL-min-1. The result shows that 6 chemical drugs shows good linear relationship (r> 0.9996) and low LOD. The method was Accurate, reliable and specificity, meet the requirements of the detection of chemical components added illegally in TCM medicines.