Simultaneous quantitative analyses of five constituents in crude and salt-processed Cuscutae Semen using a validated high-performance thin-layer chromatography method

Predominantly, the innate medicinal value of Cuscutae Semen has accounted for its extensive usage as a herbal remedy in China for centuries. Specifically, the broad applications of the medicine include aging inhibitors, anti-inflammatory agents, pain relievers, and aphrodisiacs. According to the records of crude Cuscutae Semen (CCS) and salt-processed Cuscutae Semen (PCS) in the Pharmacopoeia of the People’s Republic of China, the difference in the chemical composition between CCS and PCS is limited. In addition, an unprecedented high-performance thin-layer chromatography (HPTLC) method intended for the simultaneous quantitative determination of rutin, chlorogenic acid, hyperin, isoquercitrin, and astragalin in CCS and PCS was successfully developed. In order to achieve optimum separation, a mobile phase of ethyl acetate–formic acid–water–toluene (9:1:1:0.5, V/V ) was cautiously integrated. Moreover, the densitometric determination method was executed at specified wavelengths for various standard compounds in refection/absorption mode. In particular, the calibration curves appeared to be linear in the range of 32.64–1139.19 ng per spot for rutin, 12.94–1048.36 ng per spot for chlorogenic acid, 13.71–1110.17 ng per spot for hyperin, 1.16–297.83 ng per spot for isoquercitrin, and 2.19–710.55 ng per spot for astragalin. The proposed method is simple, precise, specific, and accurate. The statistical analysis of the data obtained proves that the method is reproducible and selective, and can be used for the routine analysis of the reported compounds in CCS and PCS samples. This study could be benefit to investigate the processing mechanism of PCS as well as support their clinical applications.