ABSTRACT
Andrographis Herba, the aerial part of Andrographis paniculata (Burm. f.) Wall. ex Nees (Acanthaceae), has a wide geographic distribution and has been used for the treatment of fever, cold, inflammation, and other infectious diseases. In markets, sellers and buyers commonly inadvertently confuse with related species. In addition, most Chinese medicinal herbs are subjected to traditional processing procedures, such as steaming and boiling, before they are sold at dispensaries; therefore, it is very difficult to identify Andrographis Herba when it is processed into Chinese medicines. The identification of species and processed medicinal materials is a growing issue in the marketplace. However, conventional methods of identification have limitations, while DNA barcoding has received considerable attention as a new potential means to identify species and processed medicinal materials. In this study, 17 standard reference materials of A. paniculata, 2 standard decoctions, 27 commercial products and two adulterants were collected. Based on the ITS2 sequence, it could successfully identify A. paniculata and adulterants. Moreover, a nucleotide signature consisting of 71 bp was designed, this sequence is highly conserved and specific within A. paniculata while divergent among other species. Then, we used these new primers to amplify the nucleotide signature region from processed materials. In conclusion, the DNA barcoding method developed in the present study for authenticating A. paniculata is rapid and cost-effective. It can be used in the future to guarantee the quality of Andrographis Herba of each regulatory link for clinical use.
Subject(s)
Andrographis , Andrographis paniculata , DNA Primers , Drugs, Chinese HerbalABSTRACT
Andrographis Herba is a commonly used plant medicine, and has been recorded in pharmacopeias of different countries. However, there are some differences in the quality standards. Based on this, this paper compare the quality standards of Andrographis Herba between Chinese Pharmacopoeia, Hong Kong Chinese Materia Medica Standards, United States Pharmacopoeia, European Pharmacopoeia and Indian Pharmacopoeia, including origin, botanical characteristics, identification(microscopic identification and chromatographic identification), content determination, specific test(such as impurities, loss on drying, extractives, pesticides, heavy metals, mycotoxins, and other items) and storage requirements, so as to provide a reference for studying international quality standards of Andrographis.
Subject(s)
Andrographis , Drugs, Chinese Herbal , Materia Medica , Reference StandardsABSTRACT
Objective: To combine macroscopical characteristic indices and chemical indices of Andrographis Herba to evaluate its quality grade. Methods: Both macroscopical characteristic indices and chemical indices (the content of four active diterpenoids and the content of ethanol-soluble extractives) of different batches of Andrographis Herba were determined. The macroscopical characteristic indices were encoded using the method of numerical taxonomy, and the content of four active diterpenoids were determined by HPLC. To screen out the appropriate indices for classification, the correlational analyses were conducted between encoded macroscopical characteristic indices and chemical indices. The quality grade was made by principal component clustering analysis according these evaluation indices, and then was analyzed through partial least squares discriminant analysis (PLS-DA). Furthermore, a partial least squares (PLS) regression was constructed for the quality grade prediction of Andrographis Herba. Results: It showed that the samples could be divided into three grades according to the principal component clustering analysis, and was reasonable evaluating by PLS-DA. The PLS regression model for quality grade of Andrographis Herba was constructed as follows: grade Y=3.761-0.020×the leaf content-0.388×the content of andrographolide-1.117×the content of neoandrographolide-0.274×the content of deoxyandrographolide-0.287×the content of 14-deoxy-11,12-didehydro-andrographolide-0.302×the content of four active diterpenoids-0.104×the content of ethanol-soluble extractives-0.015×the color of stem-0.008 4×the color of leaf-0.003×the diameter of base part of stem+0.020×the number of branch+0.137×the diameter of the upper stem+0.011×plant height, if Y=0.7-1.3, the predicted quality was grade A, if Y=1.7-2.3, then B grade, and if Y=2.7-3.3, C grade or qualified product. Conclusion: The model of grade evaluation we constructed using principal component clustering analysis combing with PLS regression analysis performed well, which was applicable in evaluating the quality grade of Andrographis Herba and other traditional Chinese medicines. It also provided a new strategy for study on grade standards of traditional Chinese medicines.
ABSTRACT
Objective: Taking electronic-eye (visual analyzer) technique,based on the powder color of Andrographis Herba,to investigate the applicability of electronic-eye technique and evaluate the quality of Andrographis Herba with different commercial specifications. Method: HPLC was employed to determine contents of andrographolide,dehydroandrographolide,14-deoxyandrographolide,neoandrographolide in 50 batches of Andrographis Herba with different commercial specifications(stems,leaves and aerial parts).Color of these samples were measured by electronic-eye technique.The data were analyzed by principal component analysis(PCA) and Pearson correlation analysis.The ability of electronic-eye to distinguish the different commercial specifications of Andrographis Herba was investigated and the correlation of chroma space system parameters (L*,a*,b*) with active components was investigated. Result: There was remarkable difference in contents of 4 diterpenoids in Andrographis Herba from different parts,their contents in leaves was the highest,followed by the aerial parts(mixture of stems and leaves),and their contents in stems was the lowest.The results of PCA was divided into two classes,namely the stem part,leaf and aerial parts,indicating that electronic-eye could be used to distinguish the quality of Andrographis Herba.The correlation results showed that there were significant negative correlation(PL*(lightness value) and the contents of andrographolide,dehydroandrographolide,14-deoxyandrographolide,neoandrographolide and the total content of these 4 components.In addition,L* of samples that did not conform to the lower limit of determination in the 2015 edition of Chinese Pharmacopeia was ≥ 69.5,and the L* of more than 90% of the samples in accordance with the requirements was Conclusion: Electronic-eye technique provides a new method and idea for the quality evaluation of Andrographis Herba.
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To solve the problems of the poor resolution of chromatographic separation,the weak durability of the relative correction factors,and the low accuracy of content determination results in the quantitative analysis of multi-components by single-marker( QAMS) method with andrographolide as the internal reference substance in the existing research of Andrographis Herba,a new QAMS method using dehydroandrographolide as the internal reference substance was established for the first time in this study. This new method can be used to simultaneously determine four diterpene lactones,including andrographolide( A),neoandrographolide( B),14-deoxyandrographolide( C),and dehydroandrographolide( S) through the optimization of chromatographic conditions and systematic investigation of methodology. At the present HPLC chromatographic conditions,four components could be well separated( R > 1. 5),and the methodology validations could satisfy the requirement of quantitative analysis. The relative correction factors( RCFs) of fA/S,fB/S,fC/S were determined as 0. 65,0. 54,0. 78,respectively. The relative standard deviations( RSDs) of their RCFs ranged between 1. 3%-5. 1%,0. 25%-0. 33%,0. 070%-0. 15%,0. 070%-0. 22%,respectively with three brands of HPLC instruments,five brands of C18 column,different flow rates( 0. 9,1. 0,1. 1 m L·min~(-1)),and different column temperatures( 25,30,35 ℃),indicating good durability of the RCFs. The relative retention value( RRV) method was used to locate the chromatographic peak of the components to be determined.The RRVs of rA/S,rB/S,and rC/Swere 0. 44,0. 86,0. 97,respectively. The RSDs of the RRVs ranged between 0. 030%-1. 6% with different HPLC instruments and columns,showing accurate peak location. The present QAMS method and the external standard method( ESM)were both used to determine the contents of four diterpene lactones from Andrographis Herba( 6 batches of medicinal materials and 18 batches of cut crude drugs). The relative errors of the determined content results between two methods were less than 2. 0%. It demonstrated that there was no significant difference in content results between these two methods,indicating good accuracy of the present QAMS method. Therefore,in this study,an accurate and highly durable QAMS method using dehydroandrographolide as the internal reference substance was established for simultaneous determination of four diterpene lactones. This method could be used to effectively control the quality of Andrographis Herba and provide technical basis for the formulation of traditional Chinese medicine industry standard and improvement of the Chinese Pharmacopoeia standard of Andrographis Herba.
Subject(s)
Andrographis , Chromatography, High Pressure Liquid , Diterpenes , Drugs, Chinese Herbal , Quality ControlABSTRACT
Objective To investigate the effects and the law of anti bitterness efficiency of hydroxypropyl-β-cyclodextrin (HP-β-CD) and other different types bitterness masking at different concentrations on the bitterness decoction of Chinese materia medica (BDCMM) by using the traditional human taste panel method (THTPM) and Electronic tongue (E-tongue). Methods Based on THTPM, the bitterness reduction value (ΔI), the correction rate of bitterness suppression (CRBS), and the correction rate of bitterness suppression potency index (PI50) were used as indicators to evaluate the bitterness suppression effect of four bitterness masking at different concentrations on BDCMM such as Sophorae Flavescentis Radix (SFR), and the relationship model between the concentrations of ΔI and bitterness masking was established to explore the bitterness suppression rule of bitterness masking on different BDCMM. Based on the E-tongue method, the E-tongue taste response information values of HP-β-CD and Acesulfame K to BDCMM such as SFR were measured at different concentrations, and the electronic tongue bitterness reduction value (ΔIe) was used as an index to explore the change rule of ΔIe with bitterness masking concentration. THTPM and E-tongue was combined to establish a prediction model of the effect of bitterness suppression. Results The ΔI, CRBS, and PI50 of four kinds of BDCMM with different concentrations of bitterness masking were measured, which can be used to compare the bitterness suppression effect of bitterness masking; The relation between ΔI and bitterness masking concentration accorded with Weibull curve model; Based on E-tongue, ΔIe was obtained, and the model of the relationship between ΔIe and bitterness masking concentration was established. The relationship between ΔIe and ΔI was established. The model determination coefficients of HP-β-CD for the two types of ΔIe of the two BDCMM were 0.986 1, 0.977 9, 0.989 0, and 0.982 0 respectively (P < 0.01, n = 6). Acesulfame had no response to the sensor and did not establish a model of bitterness suppression law. Conclusion Based on THTPM and E-tongue, a method for evaluating the bitterness suppression effect of bitterness masking on BDCMM was established. The bitterness suppression effect of bitterness masking on BDCMM such as SFR was studied by molecular inclusion, high-efficiency sweetening, and other bitterness suppression mechanisms were inverstigated.
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Process design grants the quality connotation to products. This paper was to investigate the correlation between changes of chemical fingerprints of Andrographis Herba preparation and its pharmacological activity, and set up the bridge between key process and quality attributes. By referring to the preparation process of Andrographis Herba. preparation (extracting-concentrating-drying-granulation), HPLC fingerprints were employed to determine the difference of the effective materials of the intermediate micro components. Cluster analysis results indicated that the extraction link had great influence on quality connotation variation of Andrographis Herba preparation. The pharmacological activity of various intermediates was continuously decreased in the models of DPPH antioxidant activity and LPS-induced anti-inflammatory activity in mice peritoneal macrophages. Traditional high temperature treatment process was detrimental to its clinical effect from the curve equation between the key process parameters and pharmacodynamic activity. Partial least square (PLS) was used to construct spectrum-efficiency model equation, and it was verified that this equation could accurately predict the relationship between fingerprints and pharmacological activity, which would facilitate the subsequent evaluation of quality attributes and provide scientific basis for further quality control of the whole process.
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Objective: To evaluate the feasibility of taste-masking by electronic (E)-tongue and provide a reference for the drug taste-masking study. Methods: Andrographis Herba was used as the carrier of bitter Chinese materia medica (CMM), sodium acetate (NaAc), sodium cyclamate (SC), adenosine-5′-monophosphatedisodiumalt (AMP), and 2, 4-dihydroxyhenzoic acid (DA) as the taste-masking agents. The data obtained by the E-tongue were analyzed through principal component analysis (PCA), and Euclidean distance in the data space of different dimensions was calculated. Regarding the distance of bitterness (D) as the evaluation index, then whether the data analyzed by the E-tongue and volunteers were the same or were not investigated. Results: The evaluation results of the taste-masking agents by E-tongue were similar to the taste by volunteers except NaAc, suggesting that some conditions should be required to evaluate the effect of CMM taste-masking. Conclusion: This study shows that E-tongue could be used for evaluating the effect of CMM taste-masking under certain conditions. To evaluate the gustation information by E-tongue, we should use the professional knowledge of pharmacy, chemical and other related disciplines to analyze and improve this evaluation method and model gradually.
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Objective To improve the quality standard of Andrographis Herba through determination of effective components, moisture, total ashes, acid insoluble ashes, extracts and heavy metals. Methods TLC and HPLC were used for qualitative and quantitative identification of andrographolide and dehydroandrograpolide in Andrographis Herba. Routine examinations were based on the procedures recorded in the Appendix Ⅸ A, Ⅸ H, Ⅸ K, ⅩA and Ⅸ E of Chinese Pharmacopoeia (2010), for foreign matter, moisture, ashes, extracts determination and heavy metal test respectively. Results Total content of andrographolide and dehydroandrographolide, extractives (70% ethanol) all complied with Chinese pharmacopoeia. Conclusion The established method was simple, accurate and can be used as the quality standard for the quality control of Andrographis Herba.