Multi-dimensional fingerprint profiling analysis for screening and quantification of illegal adulterated antidiabetics in hypoglycemic health products by aqueous two-phase extraction and multi-wavelength detection.

Abstract A novel method for screening and quantification of illegal adulterated antidiabetics in hypoglycemic health products was developed by multi-dimensional fingerprint profiling analysis (MDFPA). By means of aqueous two-phase extraction (ATPE), using aqueous two-phase system (ATPS) of butanol-water as the extractant, 11 common antidiabetics could be effectively extracted to the upper and lower phases, respectively. HPLC separation conditions for the extracts from two phases were investigated by multi-wavelength detection before and after p-nitrobenzoyl chloride (p-NBC) and 2,4-dinitrofluorobenzene (DNBF) derivatizations to establish multi-dimensional fingerprints. For high accuracy and reliability, a hierarchical screening approach to screening illegal adulterated antidiabetics in samples was established by MDFPA and spectral purity examination. Meanwhile, detection limits of identification for illegal adulterants were defined by detection limits of spectra (SLOD). The proposed method exhibited good identification and quantification performances. SLODs, LODs and LOQs of 11 antidiabetics were 1.22−8.37 μg/g, 0.225−4.23 μg/g and 0.755−14.10 μg/g, respectively. They had good linearity ranged from 2.0 μg/g to 300.0 μg/g (R2 ≥ 0.9978). The recoveries and RSDs were 76.83–109.6% and 0.50–6.5%, respectively. The method was successfully applied to screening of 15 batches of samples in different forms. Among them, four samples were detected to contain 5.47 μg/g of metformin, 6.50 μg/g of phenformin, 3.69 μg/g of glibenclamide and 9.11 μg/g of glimepiride, respectively. The results proved that it was an efficient and feasible alternative to screening and detection of illegal adulterated antidiabetics in hypoglycemic health products.