Development of HPLC-DAD Database for Rapid in Situ Screening of Ingredients Illegally Adulterated in Anti-Diabetic Traditional Chinese Medicines

OBJECTIVE　To develop the HPLC-DAD database of ten kinds of anti-diabetic chemicals at the condition of composition-constant mobile-phase for the rapid and in situ screening of ingredients illegally adulterated in anti-diabetic traditional Chinese medicines. METHODS　HPLC was performed on Waters XTerra MSC₁₈(4.6 mm×150 mm, 5 μm) column with two kinds of composition-constant mobile-phases: mobile phase I, acetonitrile-0.02% phosphoric acid(55︰45); and mobile phase II, acetonitrile-0.01% trifluactic acid (25︰75). The detection was performed at 229 nm with the reference wavelength of 550 nm, and DAD scanning range was 190-600 nm. At first, collect peak retention time and DAD spectrum of each reference for establishing a data library. Then, compare the peak retention time of a sample with that of the references in the database for screening suspicious substances in the sample. Finally, compare the obtained DAD spectrum of the substance with that of references in the DAD spectrum library for the further screening. Confirmation of the detected chemicals was performed by LC-MS. RESULTS　Ten kinds of anti-diabetic medicines were well separated at the above two composition constant mobile phases. The screening results of 12 batches of different anti-diabetic traditional Chinese medicine indicated that there were 5 batches containing illegally adulterated components. And four kinds of illegally added chemicals were detected in 1 batch of samples. The assay results were confirmed by HPLC-MS. CONCLUSION The assay developed here is accurate and rapid in the separation of ten kinds of anti-diabetic chemicals, and can give a strong support for the in situ screening of chemicals illegally added in anti-diabetic traditional Chinese medicine.