ABSTRACT
We have developed an HPLC-UV method for the determination of pyrroloquinoline quinone (PQQ), which utilizes a redox-based colorimetric reaction. In the proposed colorimetric reaction, the redox reaction between PQQ and dithiothreitol generates superoxide anion radicals that can convert 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (INT) to formazan dye. After PQQ separation on an octadecyl silica column, it was mixed online with dithiothreitol and INT, and the formed formazan dye was monitored by absorbance at 490 nm. The detection limit (S/N = 3) of the proposed method was 7.6 nM (152 fmol/injection). The proposed method could selectively detect PQQ in food products without any clean-up procedures.
Subject(s)
Colorimetry , Food Analysis , Fruit and Vegetable Juices/analysis , PQQ Cofactor/analysis , Chromatography, High Pressure Liquid , Molecular Structure , Oxidation-Reduction , Ultraviolet RaysABSTRACT
INTRODUCTION: Tanshinones are a major class of bioactive ingredients in the traditional herbal medicines, Danshen (Salvia miltiorrhiza). A sensitive and reliable determination method for tanshinones is useful to ensure the quality of Danshen. OBJECTIVE: To develop a sensitive and selective analytical method for tanshinones by high-performance liquid chromatography (HPLC) with fluorescence detection after pre-column derivatisation. METHODOLOGY: The proposed method depends on derivatisation reaction of tanshinones with 4-carbomethoxybenzaldehyde and ammonium acetate forming intensely fluorescent imidazole derivative. RESULTS: The proposed method provided excellent sensitivity with the detection limits of 3.3 nM (66 fmol/injection), 3.2 nM (64 fmol/injection) and 2.0 nM (40 fmol/injection) for cryptotanshinone, tanshinone I and tanshinone IIA, respectively, without the necessity of complicated instrumentations. The developed method is successfully applied to quantify the contents of tanshinones in Danshen. CONCLUSION: The developed method is the first analytical method for tanshinones by fluorescence detection. Since the derivatisation reaction is selective for the o-quinone structure of tanshinone, the developed method will become a suitable mean for the discovering of tanshinone type diterpenoids from herbal samples. Copyright © 2017 John Wiley & Sons, Ltd.