Study On Quality Evaluation Of Carthamus Tinctorius In Yunnan

Objective:The quality of the safflower in Yunnan is evaluated by using the safflower as the research object,and the HPLC and GC fingerprint of the safflower in Yunnan are established.The volatile components of the safflower and its seeds are analyzed by GC-MS.The content of the polysaccharide in the safflower is determined by the DNS method.Methods:1.Referring to the 2015 edition of Chinese Pharmacopoeia,the chemical qualitative identification of flavonoids in safflower from Yunnan were studied;water,total ash,acid insoluble ash,extract and absorbance of safflower from different areas of Yunnan were examined.2.Referring to the Chinese Pharmacopoeia of 2015 edition,the content of monosanesin in safflower from Yunnan province was determined by HPLC.Agilent ZORBAX SB-C₁₈（150 mm×4.6,5μm）column for Japanese Shimadzu High performance liquid Chromatography（LC-20A）was used,The wavelength switching was used,the wavelength of 0～15 min was 254 nm,15～45min and 360 nm was used;The mobile phase was methanol-0.2%phosphoric acid solution,the column temperature was 30℃,the sample volume was 5μL,the flow rate was 1.0 m L·min^-1,To establish a HPLC method for the simultaneous determination of Gallic acid,hydroxy safflower yellow pigment A（HSYA）,protocatechuic acid,rutin and quercetin in safflower from different areas of Yunnan.Optimization of extraction Technology of Total Sugar from Carthamus from Yunnan Province by single Factor investigation,taking the extraction rate of safflower total sugar as reference index,the effects of the ratio of material to liquid,extraction time,extraction temperature and the amount of hydrochloric acid on the extraction rate of safflower total sugar were investigated.The content of total sugar and monosaccharide in safflower was determined by3,5-dinitrosalicylic acid method,and the content of polysaccharide was calculated.3.To investigate the extraction solvent and extraction method of HPLC fingerprint of safflower from Yunnan province,optimize the chromatographic column,wavelength,column temperature,mobile phase and its gradient elution procedure,and evaluate it by traditional Chinese medicine chromatographic fingerprint evaluation system.The HPLC fingerprint of safflower was established.The extraction solvent,extraction method,extraction time,column and heating procedure of volatile components of safflower from Yunnan were studied by single factor experiment,A quartz capillary column DB-1（0.25μm×0.320mm×30 m）;FID was used,carrier gas:high purity nitrogen,inlet temperature:250℃,detector temperature:250℃;heating program:the initial temperature is 100℃,the rate of 2 min,is raised to 160℃at 10℃·min^-1,the rate of 5min,is raised to 230℃at 5℃·min^-1,and the rate of 3℃·min^-1is maintained for 10 min,and then it is raised to 250℃at the rate of 3℃·min^-1.Maintain 5 min,at the rate of 2℃·min^-1and then rise to 300℃for 3 min;The GC fingerprint of safflower was established by injection of 2μL.4.The volatile chemical constituents of safflower were analyzed by GC/MS,and the unknown compounds were identified and matched by NIST qualitative spectrum library of American National Standard Bureau and references.Results:1.The micro-identification of the safflower showed that the safflower powder contains pollen,calcium oxalate and secretory cells,and the chemical and qualitative identification results show that the safflower contains flavonoids;The content of water in the 25-batch safflower samples is 8.05～13.08%,the total ash content is 5.84～6.64%,the acid-insoluble ash is0.15～0.73%,the water-soluble extract is 37.15～56.52%,and the absorbance range is0.299～0.521.2.The content range of kaempferin in 25 batches of safflower from many producing areas in Yunnan was 0.06～4.22%.Twenty-five batches of safflower from different producing areas in Yunnan,including Gallic acid,Protocatechuic acid,Hydroxyl safflower yellow pigment a,The contents of rutin and quercetin were 1.24～3.37 mg·g^-1,7.74～34.19 mg·g^-1,61.47～97.92 mg·g^-1,11.24～83.66 mg·g^-1,1.36～19.51 mg·g^-1,respectively.The order of content was:hydroxy safflower yellow A>rutin>protocatechuic acid>quercetin>Gallic acid.3.The chromatographic conditions of HPLC fingerprint were determined as follows:column:waters symmetry?-C₁₈（150 mm×4.6,5μm）,mobile phase:methanol-0.4%phosphoric acid aqueous solution,gradient elution,detection wavelength:254 nm,flow rate:1.0 ml·min^-1,mobile phase:methanol-0.4%phosphoric acid aqueous solution,gradient elution,detection wavelength:254 nm,flow rate:1.0 ml·min^-1,Injection quantity:5μL.The GC fingerprint of essential oil of safflower from Yunnan was established.4.38 and 42 compounds were identified from the essential oil extracts of safflower and its seeds by GC/MS.Conclusion:1.The results of qualitative identification of safflower showed that there were flavonoids in safflower of Yunnan province,which was consistent with the contents of flavonoids in safflower reported in the literature;The contents of water,total ash,acid insoluble ash,water soluble extract and absorbance of 25 batches of safflower medicinal materials were determined.The results were in accordance with the Chinese Pharmacopoeia Standard 2015 Chinese Pharmacopoeia.2.The content of kaempferin in 25 batches of Yunnan safflower is within the limit of Chinese Pharmacopoeia of 2015 edition.A HPLC method was developed for simultaneous determination of Gallic acid,protocatechuic acid,hydroxy safflower yellow A,rutin and quercetin in safflower from Yunnan province.The method is simple,stable and sensitive.The established dns method for the determination of polysaccharide in safflower is simple,stable and reproducible.It can be used for quality control of safflower.3.The result of this study is one of the important means to evaluate the quality of safflower in Yunnan.The established GC fingerprints of safflower were accurate and reliable,which provided some reference for the quality control of volatile components of safflower.4.The chemical composition of volatile oil from safflower is complex.GC/MS can provide theoretical basis for the development and utilization of safflower medicinal materials and their products.