HPLC Fingerprint Establishment ,Content Determination and Multivariate Statistical Analysis of Qinlian Runfei Decoction
- VernacularTitle:芩连润肺汤的HPLC指纹图谱建立、含量测定及多元统计分析
- Author:
Kaili YAN
1
;
Chengcheng YIN
2
;
Mengyao LIU
1
;
Changsheng CUI
1
;
Ying WANG
1
;
Xiaozhou SUN
3
;
Liping SUN
3
;
Bin QI
1
;
Li LIU
1
Author Information
1. School of Pharmacy,Changchun University of TCM,Changchun 130117,China
2. Jilin Ginseng Academy,Changchun University of TCM,Changchun 130117,China
3. Children’s Diagnosis and Treatment Center,the Affiliated H ospital of Changchun University of TCM,Changchun 130117,China
- Publication Type:Journal Article
- Keywords:
Qinlian runfei decoction;
HPLC;
Fingerprint;
Content determination;
Cluster analysis;
Orthogonal partial least
- From:
China Pharmacy
2021;32(16):1956-1963
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish the f ingerprint of Qinlian runfei decoction,determine the contents of 11 components, and conduct cluster analysis and orthogonal partial least squares discriminant analysis (OPLS-DA). METHODS :HPLC method was used. The determination was performed on ZORBAX Eclipse Plus C 18 column with mobile phase consisted of acetonitrile- 0.05% phosphoric acid solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelength was set at 260 nm, and column temperature was 30 ℃. The sample size was 10 μL. Using wogonoside as reference,HPLC fingerprints of 10 batches of Qinlian runfei decoction were drawn and the similarity evaluation was conducted with Similarity Evaluation System of TCM Chromatographic Fingerprint (2012 edition),the common peaks were also confirmed ;the contents of 11 components in Qinlian runfei decoction were determined by the same method. SPSS 21.0 software was used for clus ter analysis ,and SIMCA 14.0 software was used for OPLS-DA to screen marker components affecting quality. RESULTS :There were 21 common peaks in 10 batches of Qinlian runfei decoction ,and the similarity with control fingerprint was greater than 0.98. A total of 11 common peaks were identified , which were rutin , forsythiaside A , forsythiaside B , iris, irigenin, baicalin, forsythiaside, wogonoside, baicalein, irisflorentin and wogonin. The line ar ranges of 11 components were 9.960 0-49.800 0,1.974 0-9.870 0,0.672 0-3.360 0,0.960 0-4.800 0,0.549 0- 2.745 0,5.040 0-25.200 0,1.374 0-6.870 0,0.615 0-3.075 0,0.759 9-3.795 0,0.162 0-0.810 0,0.042 0-0.210 0 μg(all r> 0.999); RSDs of precision , stability (48 h) and repeatability tests were less than 2% ; the average recoveries were 95.81%-100.29% with RSDs of 0.43%-1.73%(n=6);the contents were 8.924 4-12.820 8,0.352 2-0.868 7,0.435 6-0.711 2, 0.389 8-1.309 0,0.335 8-0.530 1,1.680 5-4.542 3,0.701 8-1.584 2,2.240 2-5.442 5,2.351 0-5.558 9,0.106 0-0.182 2,0.076 8- 0.128 9 mg/g,respectively. The results of cluster analysis showed that when class spacing was 10,it could be divided into two groups,S1-S3 and S 4-S10;when the class spacing was 5,the second class could be divided into two categories ,S6,S7,S9 were clustered into one category ,and S 4,S5,S8,S10 were clustered into one category. The results of OPLS-DA analysis showed that S6,S7 and S 9 were at the top of the figure ,S4,S5,S8 and S 10 were at the lower left side of the figure ,and S 1-S3 were at the lower right side of the figure ,which was consistent with the cluster analysis results ;VIP values of baicalin ,iris,forsythiaside A , baicalein and wogonoside were all greater than 1. CONCLUSIONS :Established fingerprint and content determination methods have high precision and good stability. Combined with multivariate statistical analysis ,it can be used for the quality control of Qinlian runfei decoction. Five components as baicalin are the marker components affecting the quality of Qinlian runfei decoction.