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Objective: 1) by hydroxy safflower yellow A (of HSYA), purification process carried screening, the establishment of a large scale preparation of. Safflower HSYA reference method, to provide the material basis for the safflower preparations and pharmacology studies. 2) to study the chemical composition of safflower essential oil. Methods: 1) gel chromatography, recrystallization, high-speed countercurrent chromatography (HSCCC) purification process to filter, respectively, from the comparison of sample throughput, yield, purity and other parameters to determine HSCCC combination of gel chromatography prepared HSYA reference substance, in accordance with the requirements of the national standard of the People's Republic of China for HSYA reference uniformity, stability testing. 2) by steam distillation and n-hexane, petroleum ether, methylene chloride three solvent extraction of safflower essential oil by gas chromatography - mass spectrometry technology (GC-MS) analysis of the chemical composition of the volatile oil. RESULTS: 1) prepared by of HSCCC and composition of the gel permeation chromatography Chromatography SYSTEM purified HSYA Reference Sample the HSCCC a solvent system n-butanol -0.1 mol / L hydrochloric acid (1:1, v / v), flow rate 1.2mL/min. The rotational speed of 850r/min, gel permeation chromatography (phi = 1:70) as mobile phase water, flow rate 1.5mL/min. From 1 g of safflower crude extract (equivalent to saffron herbs 22.86g) was isolated HSYA of Reference Sample 184 mg, purity of 99% or more in a yield of 0.805%, good uniformity, repeatability, precision, RSD less than 1%, in line with the requirements of the national standard of the People's Republic of China; 2) safflower essential oil is extracted from the steam distillation and three solvents by GC-MS analysis identified 116 kinds, the same ingredients only five kinds of content also differences were 9, 12, 15 - 18 the carbonic acid leukotriene -1 - alcohol (content of 14.27%, 11.95%, 11.23% and 2.27%, respectively), palmitic acid (13.39%, 9.93%, 6.66% and 12.46%), phyton (1.18%, 1.04%, 0.56% and 4.45%) 8,11 - 18 diene acid methyl ester (0.28%, 0.36%, 0.19% and 2.96%) and palmitic acid methyl ester (0.33 %, 0.38%, 0.11% and 1.17%). Relatively close to the chemical composition of n-hexane and petroleum ether extracts jointly to obtain the volatile oil components 21, obtained by steam distillation of the essential oil component 9 is unique, and the total content of up to 51.17%. Steam extraction and solvent extraction volatile oil is very different. Conclusion: The combination of high-speed countercurrent chromatography the gel chromatography the prepared HSYA reference, the method is stable and reliable, reproducible, high yield, simple process, applied to a large number of preparing HSYA reference and comprehensive analysis of the chemical composition of the volatile oil, red Take a volatile oil in-depth research and development to provide a reference.
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