Extraction And Separation Of Two Safflower Yellow Pigment And The Combination Lyophilized Injectable Powder Preliminary Study

Flos Carthami is the driing flower of Carthamus tinctorius L.Which is 1-year-span herb of family compositae. The Flos Carthami have the effects of promoting blood circulation and removing blood stasis analgesic,It is a traditional Chinese medicine for activating blood circulation to dissipate blood stasis. Flos Carthami is rich in Safflower yellow class components.HSYA and SYA are the key component elements. This article chooses the Flos Carthami produced in Hocheng Xinjiang Province as the research object. We study the extraction, separation, purification of the HSYA and SYA. Their combination lyophilized powder was studied in order to provide the theorical evidence to develop the source of Flos Carthami.This article first put commercial safflower injection as control,analysis the components change of Honghua injection and Flos Carthami befor and after heating by HPLC which have the high capacity of separation of complex samples. Mark out the composition of the SY family, Suggesting the need purification and low-temperature freeze-drying processof two components HYSA and SYA in Flos Carthami.This article study the method of extraction and purification of two kinds safflower yellow. Using water to extracte, macroporous resin to separate, C18 reversed-phase silica gel column and sephadex LH-20 gel column to purificate. Polyamide film and HPLC fingerprints to trace detecte. And analyzed there structures combined with spectroscopy.In line with the extraction of two SY,this article established an RP-HPLC method for simultaneous determination of Hydroxysafflor yellow-A and Safflor yellow-A in Flos Carthami. The analytical column was Akasil C18 column(4.6mm×150 mm,5μm). The mobile phase was composed of acetonitrile(A) and 0.4%phosphoric acid(B) with gradient elution (0-30min,10%A;30-50min,15%A;50-55 min,25%A;55-60min,10%A) at a flow rate of 0.7ml·min-1.The wavelength of UV detector was 403nm and column temperature was 30℃. The linear ranges of hydroxysafflor yellow-A and safflor yellow-A were 0.451-1.05μg (r=0.9993) and 0.278-0.650μg(r=0.9996) (n=5),respectively; Their average recoveries(n=6) were 100.6% and 103.0%, respectively. This method is sensitive,repeatable and suitable to determine the contents of hydroxysafflor yellow-A and safflor yellow-A in Flos Carthami.In formulation research,this article further study the lyophilized powder of HSYA and SYA. Determined the lyophilized powder Recipe by orthogonal experiments.The optimal factor for lyophilized powder Recipe is HSYA 10g, SYA 6g, mannitol 20g, EDTA-2Na 6.4mg (0.04%), pH6.0,use 0.05% carbon to decolourize, injection water to 2000mL(2mL per bottle). Do the preliminary studies of its stability and determine the analysis method.This article study the preliminary investigation of HSYA and SYA's anti-clotting and platelet aggregation.Use PH=7.4 of normal saline as a negative control, heparin as a positive control. Measured both anticoagulant and antiplatelet aggregation by in vitro experiments. The result is that HSYA and SYA both have the effect on APTT, TT, PT and anti-platelet aggregation.At the same time SYA is better than HSYA at inhibition of platelet aggregation.