Research On Identification Methods Of Traditional Chinese Medicine Based On Infrared Spectroscopy And Chemometrics

The quality of Tranditional Chinnese Medicine(TCM)has been a major problem in our country pharmaceutical industry,and the quality will directly affect its clinical efficacy.The authenticity of Chinese medicine will directly affect its clinical efficacy.The origin of Chinese medicines,the cultivation methods,the storage time and the location of medications will all affect the quality of Chinese herbal medicines.Therefore,it is urgent to identify the source and adulteration of Chinese medicinal materials more objectively and quickly and accurately.Fourier The Fourier transform infrared spectroscopy has no pollution,simple operation,small sample dosage,and can reflect the whole information of traditional Chinese medicine,not just the information of the index component.Chemometrics can be used to achieve fast and high throughput analysis of complex samples.Research Objectives:This experiment is to precious Bulbus Fritillariae Cirrhosae and the commonly used Chinese materia medica Angelica as the research object,using infrared spectroscopy combined with chemometrics methods,overall system to discuss the application of infrared spectrometry in Chinese traditional medicine quality evaluation,provide data support for establishing quality standardization of Chinese traditional medicine.Research methods:1.Using Fourier transform near infrared spectroscopy(FT-NIR)and reflective optical fiber probe,for different kinds of testing and automatic identification,fritillary bulb directly through the principal component analysis and cluster analysis to establish fritillary bulb SIMCA supervision pattern recognition model of the algorithm.2.By Fourier transform infrared spectrum of Bulbus Fritillariae Cirrhosae powder(CIR)and alcohol extract of one dimensional infrared spectroscopy,the second derivative infrared spectra and two-dimensional infrared spectrum,progressive hierarchy,fritillaria medicinal materials of complex system.Identification of the Bulbus Fritillariae Cirrhosae powder will Bulbus Fritillariae Thunbergii(THU),Bulbus Fritillariae Ussuriensis(USS),Bulbus fritillariae pallidiflorae(PAL)with 5%,10%,20%,30%,40%,50%proportion of doped in Bulbus Fritillariae Cirrhosae powder,based on the algorithm of SIMCA pattern recognition to establish the supervision of adulteration fritillary bulb model,the validation samples to evaluate the prediction accuracy of the model.Based on Partial Least Square Regression,a multi-variate calibration model was established,and the model parameters were optimized according to the measurement coefficient and standard error of prediction.3.The microscopic spectroscopic characteristics analysis of the fritillaria powder were carried out by analysis of main components and the calculation of normalised relative peak intensity method to find the microscopic infrared spectrum difference characteristics of sichuan fritillaria and other fritillaria powders.4.By using Fourier transform infrared spectrum to angelica slices of infrared spectrum analysis and the analysis of the second derivative,on the basis of the infrared spectrum correlation coefficient of angelica detection,similarity of quantitative evaluation.Research results:1.The near-infrared spectrum of the bulb of the fritillaria is similar to the starch spectrum,and the C-H extension and bending absorption peak of CIR is below 4320cm-1,but the absorption peak of PAL,THU and USS in this area is above 4320cm-1.The absorption peak of O-H stretching and C-O expansion in CIR appears near 4760cm-1,but the bands in PAL,THU and USS are close to 4750cm-1.The O-H extension absorption peak(682006790 cm-1)in CIR also appears in higher wave numbers than other kinds of fritillaria,which indicates that CIR and other kinds of fritillaria have different C-H,O-H and C-O groups.In the near infrared spectrum of the second derivative,PAL,THU and USS in the vicinity of 4860cm-1 usually peak above 4750cm-1,and the CIR only absorbs the peak at 4750cm-1.The absorption peaks near 4860cm-1 can be attributed to the combination of N-H stretching and protein amide ?,while the peaks near 4750cm-1 can be attributed to the O-H stretching and C-O expansion bands of carbohydrates.This means that PAL,THU and USS contain more protein than CIR.The principal component analysis was performed using the second derivative near infrared spectrum in the range of 6000-4000cm-1 and the first principal component could distinguish CIR and other kinds of fritillaria.The second principal component separates the PAL from the THU and the USS,while the third principal component distinguishes the PAL and USS.Then different species of fritillaria SIMCA clustering analysis,use the SNV adjusted 7400?4400cm-1 within the scope of the near infrared spectrum,SIMCA model reliability was 99%,and extension factor scores of 1.5,residual extension factor of 1.3.2.(1)infrared spectrum analysis of fritillaria powder:in the classical infrared spectrum,the absorption peak intensity at 1460cm-1,1372cm-1,1162cm-1 and 1082cm-1 can be used to distinguish wild and cultivated products.In the second derivative spectra,the absorption peak of the fritillaria in 1338cm-1 was higher than 1301cm-1,while the height of the two peaks was consistent.In the two-dimension spectra,the strongest automatic peaks of the lubei(LB),qingbei(QB)and songbei(SB)were all near 950cm-1,and the mother taibaibeimu(TB)was near 891-908cm-1,while the wabubeimu(WB)were near 990cm-1.In the CIR powder alcohol extract,the absorption peak of wild products in the range of 1800-1200cm-1 was lower than that in the range of 1200?800cm-1,while the absorption peak of the cultivated product TB and WB in the range of 1800?1200cm-1 was significantly higher than that in the range of 1200-800cm-1.In the second derivative infrared spectra,the range of 2000?200cm-1 furnace shell,green bay and songbei alcohol extract,within the range of 1570-1510cm-1 quality of wild and cultivated the peak shape of a significant difference between the two,cultivating the strongest peak is 1514cm-1,wild and strongest peak is 1561cm-1.In two-dimensional spectrum of alcohol extract of cultivating TB and WB strongest peaks automatically appear in the near and 990cm-1 and 1050cm-1,TB in 1069cm-1 and 967cm-1 automatic peak corresponds to the cross peak is positive,and they formed between apparent:"well" glyph peak cluster.(2)Fritillaria powder adulteration infrared spectrum analysis:In the classical infrared spectrum,the CIR was lower than 1464cm-1 peak,while the other kinds of fritillaria were equal to or higher than 1464cm-1 peaks in 1650cm-1.In addition,1515cm-1 was observed in the spectra of other females.Absorption peak at 1650cm-1 is O-H curve absorption band,aromatic skeleton band and amide ? band overlap,absorption peaks can be thought of as 1515cm-1 with aromatic skeleton and amide ? overlap,absorption peak in the 1464,1420 and 1372 cm-1 mainly C-H curve;The spectrum differences indicated that PAL,USS and THU contain more protein and aromatic compounds than CIR.With the increase of dopant,the absorption peak of 1650cm-1 doped CIR powder became stronger.At the same time,1515cm-1 absorption peak becomes more and more obvious.The above results show that,according to the classical infrared spectrum characteristics,the true and false,forged and adulterated CIR samples can be distinguished in the range of 1800-1200cm-1.In this research,the integrative two-dimensional correlation spectra(i2D)are defined for the first time as the multiplication of coupled synchronous and asynchronous 2D correlation spectra,which can provide correlation spectra with equal resolution but clearer features than asynchronous correlation spectra for fritillaria powder adulteration.In the region of 1200-1000 cm-1,pure CIR shows a couple of cross bands but adulterated CIR shows two couple of cross bands.Besides,as the increase of the adulterant,the cross peaks near 1400 cm-1 and 1650 cm-1 become more stronger.Therefore,the i2D correlation spectra can identify the adulterated CIR samples more intuitively than the synchronous or asynchronous correlation spectra.Finally,the FT-IR spectra in the range of 1800?1200 cm-1 are used after the SNV normalization.The reliability of the SIMCA model is 99%,while the score expansion factor is 1 and the residual expansion factor is 0.5.The total distance threshold for the recognition of true CIR is 2.024.All samples of CIR,PAL,USS?and THU can be correctly recognized as CIR or not CIR.Meanwhile,all adulterated CIR samples except for one can be correctly recognized as not CIR.The misjudged sample is the mixture of 95%CIR-Q and 5%USS.The coefficient of determination(R2)of the final PLS model is 0.987,while the validation SEP is 0.035.(3)In the microscopic spectrum of Fritillaria powder,microscopic infrared spectra of different types of Fritillaria powder,the first three principal components scored to generate different color color images,and pixels with similar spectral characteristics scored on different principal components.Closely,there are at least three types of pixels with unique spectral characteristics in different types of Fritillaria powder.The spectral characteristics of the first type of pixels are consistent with the spectral characteristics of starch and correspond to starch grains in Fritillaria powder.The second class of pixels has a higher score on the second principal component.The spectral features of this class of pixels are mainly the absorption bands of the amide I band and the amide II band around 1640cm-1 and 153 0cm-1,corresponding to the Fritillaria powder.The aleurone grains and other protein-rich microstructures.The third type of pixel scored higher on the third principal component,including C=O stretching vibration absorption near 1735cm-1,C=C stretching vibration at 1200-900cm-1,C=O stretching vibration,and sugar ring skeleton vibration absorption.Peaks,which may be derived from cellulose,hemicellulose,and lignin components that make up the cell wall,strong absorption peaks in the 1700?1500cm-1 region in Iberobia,Fritillaria and Fritillaria,suggesting that proteins and cell walls The content is relatively higher than Chuanbei.In the Fritillaria Fritillaria powder samples,the Rm value was stable at about 0.70,the standard deviation was 0.01,the Rm values of Iberoena,Pincipa and Zhebei were stable at 0.49,and the standard deviation was 0.07,which clearly distinguished Fritillaria cirrhosa.3.Different origin,storage time and medicinal parts infrared analysis:Different origin,storage time and medicinal parts infrared analysis:(1)the different regions of angelica:in the 1068cm-1 and 1053 cm-1 O-H bend stretching vibration absorption peak absorption peak height is different,the minxian(MX)and weiyuanxian(WYX)absorption peak height here have obvious difference,above 1053 cm-1 and 1068 cm-1 absorption peak;However,there is little difference in the absorption peak of zhangxian(ZX).In the second derivative spectrum,you have an aromatic ring stretching vibration absorption peak near 1514cm-1,where the peak intensity is absorbed:ZX>WYX>MX.In the vicinity of 1476cm-1 and 1460cm-1,there is C-H bending and stretching vibration peak corresponding to ester molecules,and the intensity of absorption peak is at the second place:ZX>WYX>MX.(2)different store time of angelica:storage of angelica 1 year in 1746 cm-1,1068 cm-1,990 cm-1,1053 cm-1,867 cm-1,920 cm-1 of absorption peak,stored two years of angelica on overall absorption peak intensity big,validation of sugars and glycosidic material gradually reduce along with the increase of storage time.Second derivative spectra,the storage of angelica 1 year in 1476 cm-1 C-H bending vibration absorption peak intensity slightly higher than the stock 2 years of angelica,as part of the oil characteristic absorption peak,that with the increase of storage time and the content of volatile oil in the body.(3)different medicinal parts angelica:in 1500?1200 cm-1 range,absorption peak angelica tail number is more.According to the absorbance size,the content of the chemical composition of the infrared absorption in the different medicinal parts of the angelica was:angelica tail>angelica head and whole angelica.In the second derivative range of 1800-1200cm-1,1460cm-1 and 1476cm-1 absorption peak intensity:angelica tail>angelica head>whole angelica,whole angelica absorption peak at 1408 cm-1 below the absorption peak at 1385cm-1,but angelica head and angelica tail absorption peak at 1408 cm-1 is higher than 1385cm-1.