Objective:To establish an HPLC method for the simultaneous determination of six components(uridine,adenine,adenosine(R,S)-goitrine,guanosine,clemastanin B)in Radix Isatidis,and to investigate the linear calibration with two reference substances(LCTRS)method for the qualitative analysis of multiple components in Radix Isatidis.Methods:HPLC method was used,with methanol as mobile phase A and water as mobile phase B.Gradient elution(0-3 min,3％A;3-18 min,3％A→14％A;18-25 min,14％A→26％A;25-34 min,26％A;34-40 min,26％A→46％A;40-60 min,46％A→90％A)was performed at a flow rate of 0.8 mL·min-1.The column temperature was 30 ℃,the detection wavelengths were 254 nm(0-32 min)and 230 nm(32-60 min).The injection volume was 10 μL.The actual retention time of 6 components in Radix Isatidis was determined on 13 C18 chromatographic columns of different brands and models.Guanosine and clemas-tanin B were used as double reference compounds,and LCTRS method was used to locate the chromatographic peak of each component.Three unknown chromatographic columns were used for method validation.Using guanosine as a reference substance,the relative retention time method was used to predict the retention time of the other 5 components.The predictive accuracy and column coincidence of these two methods were compared.Results:The LCTRS method could effectively predict and qualitatively analyze the retention time of six indicator components.Compared with the relative retention time method,the LCTRS method had higher accuracy in predic-ting results and better column universality.Conclusion:The LCTRS method for simultaneous determination of multiple components in Radix Isatidis is feasible and accurate,with simple operation and good durability,and has promotional value.
关键词
板蓝根/双标线性校正法/鸟苷/直铁线莲宁B/尿苷/腺嘌呤/(R,S)-告依春/腺苷
Key words
Radix Isatidis/linear calibration with two reference substances/guanosine/clemastanin B/uridine/adenine/(R,S)-goitrine/adenosine
维普
万方数据