Studies On Preparative Separation Of Reference Substance Of HSYA And Compositions Of Essential Oils Of Carthamus Tinctorius L.

Objective:1) To establish a method for the seal-up separation of reference substance, hydroxysafflower yellow A (HSYA) from Carthamus tinctorius L., the several separation methods were selected.2) The compositions of Essential oils and the three extracts by different solvents were investigated.Method:After comparing of several separation methods, HSCCC combined with Sephadex LH-20 chromatograph was comfirmed and used for the preparation of HSYA based on the sample size, yeild and purity. According to the require of national standard of China, the uniformity and stability of HSYA was tested.2) The compositions of essential oils extracted by steam distillation and three solvent extracts (n-hexane, light petroleum, dichloromethane) were investigated by GC-MS.Result:1) the HSYA was scale-up separated by the combined chromatographic methods including HSCCC and Sephadex LH-20 chromatograph. The HSCCC was performance with the two-phase solvent system composingof butanol-0.1mol/L hydrochloric acid (1:1, v/v). The flow rate was 1.2mL/min, and the rotational speed was 850r/min. The mobile phase of Sephadex LH-20 chromatograph (φ=1:70) was water with the flow rate of 1.5mL /min. the 184 mg HSYA with over 99% purity was separated from the 1g crude sample (equal to 22.86g raw material) by a single run. And yeild rate was 0.805%. Unifoemity repeatability and accuracy were good. RSD<1%, accord with the national standard.2)21 compounds from essential oils and 106 compounds from three solvent extracts were comfired by GC-MS. Among them, only 5 compounds existed in both of essential oils and extracts with various amount. They were 9,12,15-Octadecatrien-1-ol (14.27%,11.95%,11.23%和2.27%),Hexadecanoic acid (13.39%,9.93%,6.66%和12.46%) 2-Pentadecanone,6,10,14-trimethyl-(1.18%,1.04%,0.56%和4.45%) 8,11-Octadecadienoic acid, methyl ester (0.28%,0.36%,0.19%和2.96%) and hexadecanoic acid, methyl ester (0.33%.0.38%.0.11% and 1.17%), respectively. There were 21 compounds in the extracts of n-hexane and light petroleum, which were very similar.There were 9 unique compounds in the essential oils. Their content was 51.17%. the results showed that the compositions were remarkable different in the essential oils and solvent extracts.Conclusion:The method with high stability, credibility and simple of separation HSYA reference substance was established yield. the compounds of essential oils and extracts were Comprehensively analyse, which provide the reference for the future study and utilization of safflower.