Application Study On Rapid Analysis Of Main Active Components In Gegen By Near-infrared Spectroscopy

BackgroundThe quality of traditional Chinese medicine?TCM?was closely related to its clinical efficacy and safety.However,the content of active ingredients in TCM was very different due to the influence of the species,origin,planting and processing methods.In order to ensure its safety and effectiveness,it was necessary to carry out quality control of its active constituents.Traditional analysis methods,such as HPLC,GC and TLC,need complex sample pretreatment and tedious and lengthy analysis processes,so it was not suitable for quality analysis and evaluation of TCM.As near-infrared spectroscopy?NIRS?had the characteristics of nondestructive and rapid analysis,and simultaneous determination of multiple active components,as it had gradually become a new technology and attracted more and more attention for the quality evaluation of TCM.ObjectiveA rapid and non-destructive NIRS method was developed for simultaneous analysis of4 main active components of Gegen from different varieties and different habitats.The NIR methods for analysis puerarin,total flavonoids,total amino acid and total sugar was established for the quality evaluation and quality control of Gegen.This study provides theoretical basis and application support and develops the method system for the quality evaluation of TCM.MethodA total of 74 Gegen samples from 7 main geographical regions?Henan,Hebei,Guangxi,Hubei,Jiangsu,Shanxi,Sichuan,Yunnan and Anhui?and 2 varieties?Puerariae Thomsonii Radix and Puerariae Lobatae Radix?were collected and determined.The NIRS method were developed and compared based on partial least-squares?PLS?.Different model parameters like 11 kinds of spectral pre-treated method,the spectral region and the latent variables?LVs?were applied to optimize the calibrations,and the prediction set and statistical analysis was performed to test and assess the established NIRS methodResultsThe established NIRS method can be used to rapid and simultaneous analysis of 4 main active components?puerarin,total flavonoids,total amino acid and total sugar?.The NIR method for puerarin was first derivative+vector normalization(1^st D+VN)of the spectral pretreatment method,7500-4246 cm^-1 of the spectral region and 14 of the latent variables?LVs?,and the optimal model parameters:the correlation coefficients of the calibration set(R_cal)were 0.9946,root mean square errors of cross-validation?RMSECV?was 0.157,residual predictive deviation?RPD?was 13.90,root mean square error of validation,RMSEP was 0.195,correlation coefficients of the prediction set?Rpre?was 0.9922.The NIRS method for total flavonoids was min max normalization?MMN?,6100-4246 cm^-1,and 20LVs,model parameters R_cal,RMSECV,RPD,RMSEP and R_pre were 0.9959,0.335,15.60,0.507and 0.9920,respectively.The NIRS method for total amino acids was 1^st D+VN,7500-4246 cm^-1,and 17 LVs,model parameters R_cal,RMSECV,RPD,RMSEP and R_pre were0.9834,0.136,7.76,0.169 and 0.9736,respectively.The NIRS method for total sugar was1^st D+multiplicative scatter correction?MSC?,7500-4598 cm^-1,and 18 LVs,model parameters R_cal,RMSECV,RPD,RMSEP and R_pre were 0.9775,1.280,6.66,1.280 and0.9516,respectively.ConclusionThe established NIRS method was rapid,simple and accurate for the analysis of the main active components of Gegen.NIRS may be a promising method for quality analysis and control of TCM.This study provides theoretical basis and application support for the quality evaluation of TCM.