School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China.
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China; Thermo Fisher Scientific, Shanghai 201206, China.
J Chromatogr A. 2020 Jul 5;1622:461149. doi: 10.1016/j.chroma.2020.461149. Epub 2020 Apr 29.
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 (R ≥ 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.
建立了多维指纹图谱分析（MDFPA）用于筛选和定量测定降糖保健品中非法掺假的新型抗糖尿病药物的方法。通过双水相萃取（ATPE），使用丁醇-水双水相体系（ATPS）作为萃取剂，可将 11 种常用抗糖尿病药物分别有效地提取到上下相中。在进行 p-硝基苯甲酰氯（p-NBC）和 2,4-二硝基氟苯（DNBF）衍生化前后，通过多波长检测研究了从两相提取物的 HPLC 分离条件，以建立多维指纹图谱。为了提高准确性和可靠性，通过 MDFPA 和光谱纯度检查建立了一种层次筛选方法，用于筛查样品中非法掺假的抗糖尿病药物。同时，通过光谱检测限（SLOD）定义了非法掺杂物的检测限。该方法表现出良好的识别和定量性能。11 种抗糖尿病药物的 SLOD、LOD 和 LOQ 分别为 1.22-8.37 μg/g、0.225-4.23 μg/g 和 0.755-14.10 μg/g，线性范围为 2.0μg/g-300.0μg/g（R≥0.9978）。回收率和 RSD 分别为 76.83-109.6%和 0.50-6.5%。该方法成功应用于不同形式的 15 批样品的筛选。其中，4 批样品分别检测到含有 5.47μg/g 二甲双胍、6.50μg/g 苯乙双胍、3.69μg/g 格列本脲和 9.11μg/g 格列美脲。结果证明，该方法是筛选和检测降糖保健品中非法掺假的抗糖尿病药物的一种高效、可行的替代方法。
