Identification Of Stained And Weighted Safflower With Different Cultivation Areas

Safflower（the dried flower of Carthamus tinctorius L.）is a commonly used traditional Chinese medicine.It has the effects of invigorating the circulation of blood and removing obstruction in the collaterals,dissipating stasis and relieving pain.The characteristics of light weight,the red color and seeing bright are the identification index of high quality safflower,which sometimes become the counterfeit standard in the Chinese herbal medicine professional market by dyeing and weighting.As a consequence of reducing the quality and curativing effect of safflower.At the same time,safflower’s quality influenced by the place of production which is Xinjiang,Yunnan,Sichuan,Henan and other places.So,this paper applied near-infrared spectroscopy,mid-infrared spectroscopy,Raman spectroscopy and HPLC techniques to establish a method for detecting dyeing and weight-increasing safflower,and identified safflower from different producing areas to provide quality evaluation for safflower.The research content of this thesis includes the following three parts:First,Identification of stained safflower:A total of 127 safflower samples was collected from different cultivating areas and herbal markets in China to develop a rapid method to identify the dyed safflower.Near-infrared spectroscopy（NIRS）combined with characteristic identification,high performance liquid chromatography（HPLC）,principal component analysis（PCA）and partial least squares regression analysis（PLS）were employed to differentiate safflower from dyed safflower samples,and further quantify the levels of the 6 dyes,tartrazine,carmine,sunset yellow,azorubine,acid red 73 and orange II in the dyed safflower.The results indicated that the 50 safflower samples and 77 dyed safflower samples were located at different regions in PCA cluster diagram by NIR spectra.Tartrazine,carmineand and sunset yellow were found in the 77 dyed safflower samples with the amounts of0.60～3.66 mg.g^-1,0.11～1.37 mg.g^-1 and 0.10～0.71 mg.g^-1,respectively.It indicated that the three dyes were the common and main dyes in the dyed safflower.However,azorubine,acid red 73 and orange II were not detected in all herb samples.A total of62 dyed safflower samples were chosen as calibration samples to develop the model for estimating the amount of dyes in dyed safflower.The estimating accuracy was verified by another 15 dyed safflower samples.The values of tartrazine,carmine and sunset yellow in dyed safflower samples were compared between the NIRS and HPLC methods.Each value of mean absolute difference（MAD）was less than 5%.The correlation coefficients of tartrazine,carmineand and sunset yellow were 0.970,0.975 and 0.971,respectively.It indicated the data quantified by NIRS and HPLC were consistence.It is concluded that NIRS can not only differentiate safflower from dyed safflower,but also quantify the amount of the dyes.NIRS is suitable for rapidly identify the quality of safflower.Second,identification of weighted Safflower:20 safflower and 10 weighted safflower samples were collected from different origins and the Chinese herbal medicine professional market.First,the traits were identified.It was tubular flower,which was reddish yellow or red,touching soft and smelling slightly.The weighted safflower was incomplete,sticky,and touched in moist feeling.Second,total ash and acid-insoluble ash were determined.The total ash content of safflower is 9.44%（n=20）,while the total ash of weighted safflower is 29.89%（n=10）;the acid-insoluble ash of safflower is 2.43%（n=20）,weighted safflower the acid-insoluble ash was12.73%（n=10）.Further determination of the mid-infrared spectrum,the one-dimensional infrared spectrum of 20 safflowers is basically the same,the main peaks are 3367,2919～2925,2852,1731～1739,1631～1642,1515,1415～1440,1238～1245,1061～1075,833,778,628 and 535 cm^-1;and weighted safflower in calcium carbonate,with characteristic peaks of calcium carbonate(1423,876,712cm^-1);weighted safflower in potassium sulphate,Characteristic peak with potassium sulfate(1112,617 cm^-1).The Raman spectroscopy showed that the characteristic peak of calcium carbonate(1085,710,279,153 cm^-1)appeared in the weighted safflower in calcium carbonate;the characteristic peak of potassium sulfate appeared in the weighted safflower in potassium sulfate.(1143,980,610,450 cm^-1);and the safflower without weight gain has no absorption peak in the range of 100～2500 cm^-1.The contents of hydroxysafflor yellow A and kaempferol in safflower and weighted safflower were also determined by HPLC.The content of hydroxysafflor yellow A in the weight-bearing safflower is only 50%in line with the Pharmacopoeia standard,and the content of kaempferol is not up to standard;and both of the 20 safflowers meet the Pharmacopoeia standard.It is indicated that safflower and weighted safflower can be identified by these methods.Third,differentiation of safflower from different producing areas:60 safflower samples were collected from Xinjiang（20 parts）,Yunnan（20 parts）,Sichuan（10parts）,and Henan（10 parts）,and the mid-infrared spectra were determined and found to be different.The one-dimensional infrared spectrum of the safflower is different.Xinjiang safflower has obvious characteristic peaks at 877 cm^-1 and587～590 cm^-1.Sichuan safflower has characteristic peaks at 818 cm^-1,and Henan safflower has characteristic peaks at 617～619 cm^-1,while Yunnan has traits.The safflower did not find any peaks in these wave numbers.Principal component analysis of one-dimensional infrared spectroscopy showed that safflowers from different regions were distributed in four different regions.The second derivative spectra of safflower in these four producing areas are different,each with its characteristic peaks,Xinjiang safflower is at 761,743,713 cm^-1,Yunnan safflower at1442,1200,962,780 cm^-1,Henan red The flower has a characteristic peak at 1453cm^-1;the Sichuan safflower has no characteristic peak at 1680,921 cm^-1.It is indicated that the safflower of the four origins can be identified according to the peak position,peak shape,peak intensity and principal component analysis method of the one-dimensional infrared spectrum and the second derivative spectrum of the mid-infrared spectrum.