Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) method for the determination of eight anthraquinone glycosides in Rhei Radix et Rhizoma when any one of them was chosen as the internal reference substance. Methods: HPLC method was developed on a Symmetry C18 (250 mm×4.6 mm, 5 μm) column with methanol (A)-0.1% phosphoric acid (B) as mobile phase for a gradient elution. The flow rate was set at 1.0 mL·min-1, the temperature maintained at 30 ℃, and the detection wavelength was set at 280 nm. Aloe emodin-8-O-glucoside, rhein-8-O-glucoside, emodin-1-O-glucoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucoside, aloe emodin-3-CH2-O-glucoside, emodin-8-O-glucoside, and physcion-8-O-glucoside was chosen as the internal reference substance, respectively, the relative correction factors (RCFs) of other seven components to be measured to the internal reference substances were established by multi-point correction method, and the relative retention time (RRTs) of the chromatographic peaks of components to be measured to the ones of the internal reference substances were calculated, respectively. The contents of other seven components to be measured in 28 batches of Rhei Radix et Rhizoma were calculated by RCFs when one of the eight components was used as the internal reference substance respectively, and the percentage difference (PD) was used as a parameter to compare the results with that of external standard method (ESM) to verify the accuracy of QAMS. Results: When aloe emodin-8-O-glucoside, rhein-8-O-glucoside, emodin-1-O-glucoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucoside, aloe emodin-3-CH2-O-glucoside, emodin-8-O-glucoside, and physcion-8-O-glucoside was used as internal reference substance respectively, the RCFs of the other seven components to be measured to the internal reference substances were (1.435, 0.624, 1.206, 0.849, 1.718, 0.563, 0.549), (0.697, 0.435, 0.841, 0.592, 1.197, 0.392, 0.383), (1.603, 2.3, 1.932, 1.36, 2.753, 0.897, 0.88), (0.83, 1.191, 0.518, 0.704, 1.425, 0.464, 0.455), (1.178, 1.691, 0.736, 1.421, 2.024, 0.663, 0.647), (0.582, 0.835, 0.363, 0.702, 0.494, 0.328, 0.32), (1.788, 2.566, 1.117, 2.157, 1.518, 3.072, 0.982) and (1.822, 2.616, 1.137, 2.197, 1.547, 3.13, 1.025), and the RRTs were (1.207, 1.47, 1.68, 1.719, 1.763, 1.768, 2.015), (0.829, 1.218, 1.393, 1.425, 1.461, 1.466, 1.671), (0.681, 0.821, 1.143, 1.169, 1.199, 1.203, 1.371), (0.596, 0.719, 0.876, 1.023, 1.049, 1.053, 1.2), (0.583, 0.703, 0.856, 0.977, 1.026, 1.029, 1.173), (0.568, 0.685, 0.835, 0.953, 0.975, 1.003, 1.143), (0.566, 0.683, 0.832, 0.95, 0.972, 0.997, 1.139) and (0.497, 0.599, 0.73, 0.834, 0.853, 0.875, 0.878), respectively. The PD between the mean values of the contents of eight components calculated by QAMS and the results determined by ESM were less than 3%. Conclusion: The established QAMS method is accurate and feasible, and can simultaneously determine eight anthraquinone glycosides in Rhei Radix et Rhizoma with only one reference substance in practical work.
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