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ObjectiveTo analyze the quantity-quality transfer of standard decoction of Ginseng Radix et Rhizoma(GRR) decoction pieces produced by fresh and traditional cutting, and to provide reference for quality control and application development of the decoction pieces produced by fresh cutting. MethodTen batches of representative GRR decoction pieces produced by fresh and traditional cutting and their standard decoctions were prepared by standard process, and high performance liquid chromatography(HPLC) fingerprint of the standard decoction was established and performed on an Agilent EC-C18 column(4.6 mm×150 mm, 2.7 μm) with acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) as the mobile phase for gradient elution(0-23 min, 18%-21%A; 23-35 min, 21%-28%A; 35-80 min, 28%-32%A), and the detection wavelength was 203 nm. Then similarity evaluation, principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) of fingerprint of the standard decoction were performed to screen the differential components with variable importance in the projection(VIP) value>1. Quantitative analysis was carried out on the screened known differential components, and combined with the indicators of the dry extract rate and the transfer rate, to explore the differences in the quantity-quality transfer between the standard decoction of GRR decoction pieces produced by fresh and traditional cutting. ResultThe fingerprint similarity of the standard decoction of GRR decoction pieces produced by fresh and traditional cutting was more than 0.950, and 18 common peaks were identified, including 9 identified common peaks. The results of PCA and PLS-DA showed that there were some differences in the contents of index components between the two standard decoctions. The contents of ginsenoside Rg1, Re and Ro in GRR decoction pieces produced by fresh cutting were higher than those in traditional decoction pieces, while the contents of ginsenoside Rb1, Rc , Rb2 and Rd were lower than those in traditional decoction pieces. The contents of ginsenoside Rg1, Re, Rb1 and Ro in the standard decoction of GRR decoction pieces produced by fresh cutting were higher than those in the standard decoction of traditional decoction pieces, while the contents of ginsenoside Rc , Rb2 and Rd were comparable between the two standard decoctions. Compared with the standard decoction of the traditional decoction pieces, the average transfer rates of ginsenoside Rg1, Rb1, Rc, Rb2 and dry extract rate of the standard decoction of GRR decoction pieces produced by fresh cutting were significantly increased(P<0.05), and the average transfer rate of ginsenoside Re and Rd also increased, but the difference was not statistically significant. ConclusionThe dry extract rate, content and transfer rate of index components of standard decoction of GRR decoction pieces produced by fresh cutting are better than those of the standard decoction of traditional decoction pieces, which can provides data support for the subsequent clinical application of fresh cutting products.
ABSTRACT
ObjectiveTo analyze the quantity-quality transfer of standard decoction of Ginseng Radix et Rhizoma(GRR) decoction pieces produced by fresh and traditional cutting, and to provide reference for quality control and application development of the decoction pieces produced by fresh cutting. MethodTen batches of representative GRR decoction pieces produced by fresh and traditional cutting and their standard decoctions were prepared by standard process, and high performance liquid chromatography(HPLC) fingerprint of the standard decoction was established and performed on an Agilent EC-C18 column(4.6 mm×150 mm, 2.7 μm) with acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) as the mobile phase for gradient elution(0-23 min, 18%-21%A; 23-35 min, 21%-28%A; 35-80 min, 28%-32%A), and the detection wavelength was 203 nm. Then similarity evaluation, principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) of fingerprint of the standard decoction were performed to screen the differential components with variable importance in the projection(VIP) value>1. Quantitative analysis was carried out on the screened known differential components, and combined with the indicators of the dry extract rate and the transfer rate, to explore the differences in the quantity-quality transfer between the standard decoction of GRR decoction pieces produced by fresh and traditional cutting. ResultThe fingerprint similarity of the standard decoction of GRR decoction pieces produced by fresh and traditional cutting was more than 0.950, and 18 common peaks were identified, including 9 identified common peaks. The results of PCA and PLS-DA showed that there were some differences in the contents of index components between the two standard decoctions. The contents of ginsenoside Rg1, Re and Ro in GRR decoction pieces produced by fresh cutting were higher than those in traditional decoction pieces, while the contents of ginsenoside Rb1, Rc , Rb2 and Rd were lower than those in traditional decoction pieces. The contents of ginsenoside Rg1, Re, Rb1 and Ro in the standard decoction of GRR decoction pieces produced by fresh cutting were higher than those in the standard decoction of traditional decoction pieces, while the contents of ginsenoside Rc , Rb2 and Rd were comparable between the two standard decoctions. Compared with the standard decoction of the traditional decoction pieces, the average transfer rates of ginsenoside Rg1, Rb1, Rc, Rb2 and dry extract rate of the standard decoction of GRR decoction pieces produced by fresh cutting were significantly increased(P<0.05), and the average transfer rate of ginsenoside Re and Rd also increased, but the difference was not statistically significant. ConclusionThe dry extract rate, content and transfer rate of index components of standard decoction of GRR decoction pieces produced by fresh cutting are better than those of the standard decoction of traditional decoction pieces, which can provides data support for the subsequent clinical application of fresh cutting products.
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ObjectiveTo analyze and determine the differential components of freeze-dried and sun-dried Panacis Quinquefolii Radix(PQR), and to compare the differences in their pro-angiogenic activities. MethodFingerprints of freeze-dried and sun-dried PQR were established based on ultra performance liquid chromatography(UPLC), and chemometrics methods such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were combined to determine the differential saponin composition of the two decoction pieces, and six representative saponins were selected and their contents in freeze-dried and sun-dried PQR were determined by UPLC. Transgenic zebrafish line Tg(fli1a∶EGFP) embryos fertilized for 24 h were selected, and different doses of 70% methanol extracts of freeze-dried and sun-dried PQR(10, 30 mg·L-1) were used to intervene in normal zebrafish and in a zebrafish model of intersegmental vascular(ISV) injury induced by vascular endothelial growth factor(VEGF) receptor tyrosine kinase inhibitor Ⅱ(PTK787), then the development of subintestinal vein(SIV) and ISV of zebrafish was observed, SIV diameter, mean number of crossings and mean number of germinations were determined, and the ISV vascular index was calculated, in order to compare the pro-angiogenic activities of the two decoction pieces. ResultThe similarity of the fingerprints of freeze-dried and sun-dried PQR decoction pieces was>0.950, and 17 common peaks were identified, of which 6 common peaks were designated as peak 6(ginsenoside Rg1), peak 7(ginsenoside Re), peak 8(ginsenoside Rb1), peak 11(ginsenoside Rc), peak 13(ginsenoside Rb2), and peak 16(ginsenoside Rd), respectively. A total of 11 differential saponin components were screened by PCA and OPLS-DA, indicating that there were some differences in the contents of the components in the two decoction pieces. The results of determination showed that the contents of ginsenosides Rg1, Re, Rb1 and Rb2 in freeze-dried PQR were higher than those in sun-dried PQR, while the contents of ginsenosides Rc and Rd were lower than those in sun-dried PQR(P<0.05, P<0.01). In the study of the pro-angiogenic effect on normal zebrafish embryos, compared with the blank group, and the SIV vessel diameter, mean germination rate and mean crossover rate were significantly higher in the high-dose groups of freeze-dried and sun-dried PQR(P<0.01), and the vessel diameter, mean numbers of crossings and germinations in the freeze-dried PQR group were higher than those of the sun-dried PQR group(P<0.05). In the study of the pro-angiogenic effect on zebrafish embryos with ISV injury, the development of ISV in the model group was significantly inhibited when compared with the blank group, compared with the model group, different dose groups of freeze-dried and sun-dried PQR could promote the growth and sprouting of ISV, and the number of normal blood vessels in the freeze-dried PQR group was significantly higher than that in the sun-dried PQR group at the same dosage(P<0.05). ConclusionFreeze-drying can effectively avoid the loss and secondary transformation of ginsenosides in PQR, and its angiogenic activity is better than that of sun-dried PQR, which can provide a reference for the production and development of high-quality PQR decoction pieces.
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OBJECTIVE To establish the grade s tandard for Panax quinquefoli um and to evaluate the quality of different grades of medicinal materials. METHODS Totally 24 batches of P. quinquefolium were used as test samples. Pearson correlation analysis method was used to analyze the correlation between qualitative analysis indicators (taproot length ,taproot diameter and weight of single root )and internal component indicators (ethanol-soluble extract ,and the contents of ginsenoside Rg 1,ginsenoside Re , ginsenoside Rb 1,ginsenoside Rc ,ginsenoside Rb 2,ginsenoside Rd ,pseudo-ginsenoside F 11). Combined with chemometrics methods,the reference indexes for the classification of P. quinquefolium were selected ,and the classification standards were formulated. HPLC-ELSD fingerprints of 24 batches of P. quinquefolium were established and their similarity evaluation was also performed. The chromatographic peaks were identified by comparison with the reference substance ,and then the quality of different grades of P. quinquefolium was evaluated by cluster analysis. RESULTS After screening ,taproot diameter ,the weight of single root and the content of ginsenoside Rd were taken as the reference indexes for the classification of P. quinquefolium . According to above 3 indexes,P. quinquefolium were divided into 3 grades:special grade ,first grade and second grade. According to the center value of K-means clustering ,the total score of special-grade medicinal materials was more than 135.40,that of first-grade medicinal materials was 61.82-135.40,and that of second-grade medicinal materials was less than 61.82. In the HPLC-ELSD fingerprints of 24 batches of P. quinquefolium ,25 common peaks were confirmed ,and 7 characteristic peaks were identified. The similarity of the chromatograms of P. quinquefolium of special grade ,first grade and second grade with fingerprints ranged 0.980-0.989,0.962-0.968,0.940-0.949,respectively. The results of cluster analysis showed that different grades of P. quinquefolium could be identified significantly. CONCLUSIONS The grade standard and HPLC-ELSD fingerprints of P. quinquefolium are established,which can be applied for exclusive identification of P. quinquefolium ,and provide reference for its quality control and grade classification.
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OBJECTIVE:To establish a method for the contents determination of phospholipid and γ-linolenic acid in Com-pound linolenic acid soft capsule. METHODS:HPLC was conducted to determine the content of soybean lecithin;the column was Agilent TC-C18 with mobile phase of methanol-water(84:16,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 219 nm,the column temperature was 30 ℃,and the injection volume was 10 μl.Gas chromatography was conducted to determine the content of γ-linolenic acid in the preparation;the column was DB WAX,injector temperature was 210 ℃ by temperature pro-grammed,carrier gas was helium at a flow rate of 1 ml/min by split injection(split ratio of 1:30),and the injection volume was 0.02μl. RESULTS:The linear range was 4.64-16.24μg/ml for phospholipid(r=0.999 6)and 0.093 44-0.327 04μg/ml forγ-linole-nic acid methy ester(r=0.999 6);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 97.44%-99.36%(RSD=0.93%,n=6) and 97.22%-99.07%(RSD=1.01%,n=6),respectively. CONCLUSIONS:The method is simple,stable with good separation,and can be use for the contents determination of phospholipid and γ-linolenic acid in Com-pound linolenic acid soft capsule.
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OBJECTIVE:To establish the HPLC fingerprint for Anemone raddeana. METHODS:HPLC was performed on the column of Phemomenex Gemini C18 with mobile phase of 0.1%phosphoric acid-acetonitrile(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 206 nm,the column temperature was 30℃,and the injection volume was 20μl. With the refer-ence of raddeanin A,13 batches of A. raddeana were analyzed,chromatographic fingerprint similarity evaluation system software was conducted for similarity analysis,and SPSS 13.0 was conducted for cluster analysis. RESULTS:There were 11 common peaks in the 13 batches of A. raddeana with similarity of higher than 0.90. According to the verification,the fingerprint and control fin-gerprint shows good consistency. The drugs in Huadian,Jiaohe, Tiangang,Shulan,Tonghua and Fusong of Jilin and Shangzhi of Heilongjiang were regarded as category 1,and in Harbin,Yabuli town and Yimianpo of Heilongjiang,Qingyuan of Liaoning,Ji-nan of Shandong were category 2. CONCLUSIONS:The established fingerprint can provide reference for the identification and quality evaluation of A. raddeana.