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Objective To establish a rapid determination model based on near-infrared spectroscopy (NIRS) for glycyrrhizic acid and liquiritin in Glycyrrhizae Radix et Rhizoma Yinpian. Methods The contents of glycyrrhizic acid and liquiritin in Glycyrrhizae Radix et Rhizoma Yinpian from different places of origin were determined by high performance liquid chromatography (HPLC) as reference values. At the same time 2 200-2 049, 1 750-1 450, 1 151-1 001 nm and 1 795-1 475, 1 395-1 293, 1 125-1 030 nm wavelength ranges of near-infrared spectra were selected to establish the rapid determination model by combining partial least squares (PLS) regression analysis with cross validation method. Results The correlation coefficient and root-mean-squares error of cross validation of the established content calibration model were 0.980 and 0.184 for glycyrrhizic acid, and 0.919 and 0.144 for liquiritin, respectively. Conclusion The NIRS-PLS method is convenient, rapid and nondestructive for the content determination of glycyrrhizic acid and liquiritin for large number of Glycyrrhizae Radix et Rhizoma Yinpian, which provides a new and feasible method for the rapid quality evaluation of Glycyrrhizae Radix et Rhizoma Yinpian.
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Objective: Based on response surface methodology, HPLC was applied to quantitatively determine the optimal processing technology of Glycyrrhizae Radix et Rhizoma Praeparata cum Melle (GRRPM) from the perspective of multi-index and comprehensive evaluation. Methods: HPLC was used for quantitative analysis, and the content of liquiritin, liquiritigenin, licochalcone A and glycyrrhetinic acid was used as inspection indexes. Response surface methodology was used to investigate the effects of the adding amount of honey, steaming and soaking time, frying temperature and frying time on the processing technology of GRRPM, and to optimize the optimal processing technology of GRRPM. Results: The chromatographic column was Diamonsil C18 (2) (4.6 mm × 200 mm, 5 μm); mobile phase was acetonitrile-0.1% phosphate aqueous solution, gradient eluting: 0-20 min, 12%-32% acetonitrile; 20-45 min, 32%-70% acetonitrile; 45-75 min, 70%-97% acetonitrile, with detection wavelength of 260 nm, column temperature of 20 ℃, and flow rate of 1 mL/min; Using liquiritin as internal standard, the relative correction factors of glycyrrhizin, licochalcone A, glycyrrhizinic acid and their relative correction factors were determined and calculated to be 0.56, 0.64 and 1.42, respectively. The optimum processing process of GRRPM was as follows: the amount of honey was 1/4, the soaking time was 15 min, frying pan bottom temperature was 160 ℃, and frying time was 13 min. Conclusion: The results of systematic adaptability investigation of the experimental content determination method meet the requirements. The best processing scheme of GRRPM optimized by response surface methodology is feasible and provides scientific basis for formulating quality standards and modern research of GRRPM.
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Objective: To make a preliminary prediction of the Q-marker of Glycyrrhizae Radix et Rhizoma from the perspective of the effectiveness and measurability of chemical components based on the concept of Q-marker of Chinese materia medica. Methods: Based on literature integration and data analysis, the source range of Glycyrrhizae Radix et Rhizoma Q-marker was screened, and the effectiveness of the ingredients was analyzed through network pharmacology. Qualitative and quantitative analysis of 15 batches of Glycyrrhizae Radix et Rhizoma from four places of origin was performed by HPLC. The pattern recognition method was used to screen out the main marker components that caused the differences between groups, which were combined with network pharmacological results to further determine the Q-marker of Glycyrrhizae Radix et Rhizoma. Results: Literature studies had determined that flavonoids and triterpenoids were the main source of Glycyrrhizae Radix et Rhizoma Q-marker; Network pharmacology results showed that liquiritin, glycyrrhizic acid and other components had high connectivity in the "component-target-pathway" network and were the main active components; The fingerprints of 15 batches of Glycyrrhizae Radix et Rhizoma samples were established, and five components, including liquiritin and liquiritin apioside, were identified as the main marker components by PLS-DA analysis; The content determination results of liquiritin, liquiritin apioside, glycyrrhizic acid and glycyrrhetinic acid showed that there were significant differences in the content of ingredients among different production areas. The qualitative and quantitative research on pharmacology combined with network pharmacology revealed that liquiritin, liquiritin apioside, glycyrrhizic acid and glycyrrhetinic acid can be used as Glycyrrhizae Radix et Rhizoma Q-marker. Conclusion: Taking flavonoids and triterpenoids as the source of Q-marker for Glycyrrhizae Radix et Rhizoma, the qualitative and quantitative (measurability) study of Glycyrrhizae Radix et Rhizoma herbs from multiple producing areas combined with network pharmacology (effectiveness) revealed liquiritin, liquiritin apioside, glycyrrhizic acid and glycyrrhetinic acid as the potential Q-marker of Glycyrrhizae Radix et Rhizoma are scientific and reasonable, which provide reference for quality control of Glycyrrhizae Radix et Rhizoma.
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Objective: To establish the HPLC fingerprint and the determination method of three index components of the classical herbal formula substance benchmarks of Xiebai Powder. Methods: Fingerprint chromatographic conditions were as following: detection wavelength 254 nm/325 nm, column temperature 35 ℃; flow rate 0.8 mL/min; injection volume 25 μL; mobile phase consisting of 0.1% aqueous formic acid (A) and acetonitrile (B); binary gradient elution: 0-20 min, 5%-10% B; 20-33 min, 10%-15% B; 33-50 min, 15%-20% B; 50-95 min, 20%-58% B. Ten batches of substance benchmarks of Xiebai Powder fingerprints were collected and evaluated by the Chinese Pharmacopoeia Committee "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System 2012 Edition" software. Chromatographic conditions of content determination: detection wavelength 237 nm, column temperature 30 ℃; flow rate 1.0 mL/min; injection volume 5 μL; mobile phase consisting of 0.1% aqueous phosphoric acid (A) and acetonitrile (B) for binary gradient elution: 0-10 min, 5%-20% B; 10-18 min, 20%-60% B; 18-26 min, 60%-100% B; 26-38 min, 100% B; 38-41 min, 100%-5% B; 41-45 min, 5% B. Results: Based on the matching results, 55 common peaks were determined at a wavelength of 254 nm, and 57 common peaks were determined at a wavelength of 325 nm. Three substances, mulberroside A (S), liquiritin and ammonium glycyrrhizinate, were identified in the common peaks. After methodological research, its precision, stability and reproducibility were good. Ten batches of substance benchmarks of Xiebai Powder fingerprints were evaluated with reference fingerprints, and their similarities were greater than 0.9. The average recovery rates of mulberroside A (S), liquiritin and ammonium glycyrrhizinate were 97.82%, 97.40% and 105.81%, respectively. The RSD (n = 6) was 4.41%, 2.51% and 1.19%, respectively, which met the require of 2015 edition of the Chinese Pharmacopoeia. The three components had good linearity in the range of 25.25-2525 ng, 25-2 500 ng and 8.5-850 ng, respectively. The method had good precision, stability and repeatability. The contents of 10 batches of substance benchmarks of Xiebai was determined. The content of mulberry A was 11.6-35.5 mg/g, the content of liquiritin was 0.1-1.6 mg/g, and the content of glycyrrhizic acid was 0.3-2.5 mg/g. The range of the contents of these ingredients was large, which indicated that the quality of mulberry husks and licorice herbs from different places was quite different. Conclusion: The establishment of the HPLC fingerprint and the determination method of three index components of the classical herbal formula substance benchmarks of Xiebai Powder provided some bases for the study of the quality standard of substance benchmarks of Xiebai Powder.
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Objective::To develop high performance liquid chromatography-diode array detector (HPLC-DAD) wavelength switching for simultaneously determining the contents of inosine, loganic acid, chlorogenic acid, amygdalin, hydroxysafflor yellow A, gentiopicroside, ferulic acid and liquiritin in 15 batches of material benchmarks of Shentong Zhuyutang. Method::The quantitative analysis was carried out on a Thermo Hypersil GOLD C18 column (4.6 mm×250 mm, 5 μm) with mobile phase of acetonitrile-0.1%phosphoric acid aqueous solution for gradient elution, the flow rate was 1.0 mL·min-1, the detection wavelengths were set as 248 nm (0-11 min, inosine), 235 nm (11-14 min, loganic acid), 324 nm (14-16 min, chlorogenic acid), 220 nm (16-19 min, amygdalin and hydroxysafflor yellow A), 274 nm (19-26 min, gentiopicroside), 247 nm (26-54 min, ferulic acid and liquiritin), the column temperature was maintained at 25 ℃. According to the contents of eight active components in 15 batches of material benchmarks, orthogonal partial least squares discriminant analysis (OPLS-DA) in SIMCA 14.1 was used to evaluate the quality difference of each batch of samples. Result::Each component had good separations, the linear ranges of the above 8 components were 2.1-67.2, 1.812 5-58, 1.937 5-62, 5.212 5-166.8, 8.45-270.4, 7.075-226.4, 1.775-56.8, 3.875-124 mg·L-1, respectively (r≥0.999 6). The average recoveries of them were 99.23%, 100.09%, 99.33%, 98.85%, 99.15%, 98.75%, 99.42%, 98.96%, respectively (RSD<2%). The contents of the above eight components in 15 batches of material benchmarks were 0.183 5-0.250 3, 0.173 1-0.265 3, 0.069 5-0.169 8, 0.959 2-1.458 2, 1.905 4-2.553 3, 0.933 3-1.997 5, 0.084 6-0.143 4, 0.212 5-0.704 3 mg·g-1, respectively. Liquiritin, ferulic acid, gentiopicroside and hydroxysafflor yellow A were determined to have significant impact on the quality of different batches of material benchmarks of Shentong Zhuyutang through OPLS-DA. Conclusion::The established method for simultaneous determination of multi-components is reliable, simple and in line with the requirements of methodological verification. It is suitable for the quality control of research and development of compound preparations of Shentong Zhuyutang.
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Objective:To identify the quality differential markers of different processed products of Glycyrrhiza uralensis dry roots and rhizomes. Method:Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MSE) was used to collect high-precision mass-charge ratio and ion response strength information of the components in G. uralensis dry roots and rhizomes before and after processing by negative ion mode. The data set collected after pretreatment was analyzed with principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) to quickly search the differential components in different processed products of G. uralensis dry roots and rhizomes. Differential components were identified according to the relative molecular weight, fragment ion, mass spectrum database and related literature information, then the migration of components before and after processing was studied. Result:A total of 10 quality differential markers were searched from raw products, roasted products and honey-roasted products of G. uralensis dry roots and rhizomes, mainly derivatives of liquiritin and glycyrrhizic acid. Among them, the contents of 6''-O-acetylliquiritin apioside, 6''-O-acetylliquiritin apioside isomer, 6''-O-acetylliquiritin, formononetin and 11-deoxo-18β-glycyrrhetic acid were the highest in the raw products, the contents of 6''-O-acetylisoliquiritin apioside, 6''-O-acetylisoliquiritin, isoliquiritin and glycyrrhetic acid 3-O-glucuronide were the highest in the roasted products, the content of liquiritin was the lowest in the honey-roasted products. Conclusion:There are some chemical differences among the three products. This study can provide material basis for the quality control and pharmacodynamic research of processed products of G. uralensis dry roots and rhizomes.
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Objective:To establish the HPLC fingerprint test method for the medicinal materials, decoction pieces and substance benchmarks of Shaoyao Gancaotang for investigating the quality transmitting of substance group in preparation process of the medicinal materials-decoction pieces-substance benchmarks, then to evaluate the scientificity and rationality of preparation process of substance benchmarks of Shaoyao Gancaotang by combining with yields of dry extract, transfer rates of effective components and other indexes. Method:Substance benchmarks of Shaoyao Gancaotang was prepared according to the method recorded in ancient medical books, fingerprints of 15 batches of medicinal materials, decoction pieces and substance benchmarks were detected by HPLC, and the contents of effective ingredients were determined. At the same time, the correlation analysis of quality transmitting of substance group during the preparation of substance benchmarks was carried out by combining the yields of dry extract and transfer rates of effective components. Result:The established HPLC fingerprint method has good precision, repeatability and stability, it can be used for the simultaneous determination of fingerprint of medicinal materials, decoction pieces and substance benchmarks. In the fingerprint of substance benchmarks, 16 common peaks were determined by taking liquiritin as the reference peak, of which 6 chromatographic peaks belong to Paeoniae Radix Alba and 11 chromatographic peaks belong to Glycyrrhizae Radix et Rhizoma, 7 major chromatographic peaks were identified. The similarities of fingerprints of 15 batches of medicinal materials, decoction pieces and substance benchmarks of Shaoyao Gancaotang were good by comparing with their respective reference fingerprints(≥ 0.90), the average dry extract rate of 15 batches of substance benchmarks was 24.81%, and no discrete data were found; the average transfer rates of paeoniflorin, liquiritin and glycyrrhizic acid from decoction pieces to substance benchmarks were 79.68%, 63.70% and 51.20%, respectively, and no discrete data were found. Conclusion:In this paper, a scientific and reasonable method for evaluating the process of substance benchmarks of Shaoyao Gancaotang is established by means of the fingerprint method controlled by the whole substance group, the research idea of quality transmitting of substance group in the preparation process, and the evaluation of technical and economic indicators. It can be used as a reference for the evaluation and research of material benchmarks in other famous classical formulas.
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Objective Based on the main morphological traits and chemical constituents of 42 Glycyrrhiza uralensis germplasm in China, the genetic diversity of them was analyzed comprehensively. Methods Twelve morphological traits and five kinds of chemical components of G. uralensis germplasm transplanted in the same place were collected. The genetic diversity index and coefficient of variation were calculated, using cluster analysis and principal component analysis for statistical analysis. Results Among the five chemical constituents, the highest genetic diversity index of liquiritin content was 2.05; The maximum coefficient of variation of isoliquiritin content was 99.50%; The content of liquiritin was moderately correlated with the content of isoliquiritin. Among 12 morphological traits, the highest genetic diversity index of plant height was 2.08, and the maximum coefficient of variation of actual fruit sequence was 37.09%. The variation of fruit type and plant type was greater than that of leaf type. Cluster analysis divided 42 germplasms into three types, and the second group had better germplasm quality. The principal component analysis reduced 17 indicators to six factors, with a cumulative contribution rate of 72.96%. Factor 6 was a factor that represents glycyrrhizic acid, liquiritin, and isoliquiritin. Conclusion The genetic diversity of the main morphological traits and chemical constituents of 42 G. uralensis germplasms is rich, and six excellent germplasms are V08, V10, V17, V34, V36, and V38.
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Objective To establish a complexation extraction and back extraction technology for the separation and purification of liquiritin from Glycyrrhiza uralensis ultrafiltrate. Methods Taking the extraction rate of liquiritin as an index, the optimum composition of complexing extractant was first determined by uniform design, and then orthogonal experiments were used to optimize the conditions of complexing extraction. Taking the back extraction rate of liquiritin as an index, the process conditions for the back extraction of liquiritin were determined by investigating the type and concentration of back extractant. Results The complexation extraction research found that the complexing extractant should be a binary complexing extractant composed of trialkyl phosphine oxide (TRPO) and sulfonated kerosene. The optimum extraction conditions for liquiritin were as follows: TRPO-sulfonated kerosene (9∶91), pH value of G. uralensis ultrafiltrate was adjusted to 4, volume ratio of organic phase to aqueous phase was 1∶1, and average extraction rate of liquiritin reached 99.6%. The study of back extraction process showed that under the condition that the volume ratio of organic phase to back extractant was 1∶1, 17.5 mmol/L NaOH aqueous solution was the best back extractant, and the back extraction rate of liquiritin was 99.3%. Conclusion Under the optimized conditions, the liquiritin in G. uralensis ultrafiltrate can smoothly transfer from the ultrafiltrate to the complexing extractant and then to the alkaline back extractant. The total transfer rate of liquiritin is as high as 98.9%. This paper can provide a new preparation technology for the separation and purification of liquiritin.
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Objective To investigate the effects of the three methods of decocting with deslag, decocting without deslag, and double decocting on the content of nine ingredients baicalin, baicalein, ginsenoside Re, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, liquiritin, 6-gingerol, berberine hydrochloride, palmatine hydrochloride, and total flavonoids in Banxia Xiexin Decoction (BXD). Methods Nine index components were determined by HPLC. The HPLC analysis was performed on Welch Ultimate XB-C18 column (250 mm × 4.6 mm, 5 μm) with mobile phase of acetonitrile-0.1% phosphate aqueous solution for gradient elution; And carried out at column temperature of 28 ℃, volume flow of 0.9 mL/min, and detection wavelength of 203, 252, 280, and 355 nm. The total flavonoids were determined by colorimetry. Results Nine kinds of ingredients and total flavonoids could be detected in three different decoctions. In the method of decocting with deslag, baicalin, baicalein, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, and liquiritin increased by 10.01%, 12.88%, 29.09%, 16.75%, and 15.02%, respectively, compared with decocting without deslag; It decreased by 5.54%, 4.15%, 14.49%, 7.85%, and 9.18%, respectively compared with double decocting; Ginsenoside Re, 6-gingerol, berberine hydrochloride, and palmatine hydrochloride increased by 37.90%, 3.78%, 5.33%, and 5.99% compared with decocting without deslag, respectively; compared to the double decocting methods, it increased by 1.07%, 11.57%, 3.41%, and 1.93%. The total flavonoids increased 22.61% higher than decocting without deslag and 6.54% higher than double decocting. Conclusion: The results can effectively reflect the quality difference of different decocting methods. Among the three methods of decoction, the method of decocting without deslag has significantly improved the dissolution of the active ingredients of each component in the decoction, and improve the clinical efficacy of BXD to a certain extent. It provides a good experimental basis for the decocting without deslag method used in Zhang Zhongjing’s Treatise on Febrile Diseases.
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Objective: To establish a research strategy for discovering quality marker (Q-marker) of Shenzhiling Oral Liquid based on the “fingerprint-efficacy-pharmacokinetics” correlation. Methods: HPLC fingerprints and acetylcholinesterase (AchE) inhibitory activities of 12 batches of Shenzhiling Oral Liquid were analyzed. The correlation analysis between the HPLC fingerprints and AchE inhibitory effects were carried out with orthogonal signal correction-partial least squares regression (OSC-PLSR) method. Combined with network pharmacology, efficacy-related components were determined. By identifying the compounds absorbed and exposed in vivo, pharmacologically active components were determined. Finally, Q-marker could be preliminarily discovered by the comprehensive analysis associated with the integration of efficacy-related components and pharmacologically active ingredients. Results: The results of OSC-PLSR analysis showed that three efficacy-related components were closely related to AchE inhibitory activities. According to mapping the targets of diseases, 61 efficacy-related components were determined. Eleven active compounds in plasma were identified by UHPLC-quadrupole-orbitrap-MS. The Q-markers of Shenzhiling Oral Liquid were liquiritin apioside, albiflorin and azelaic acid preliminarily determined by integrated and comprehensive analysis. Conclusion: The combination of fingerprint-efficacy relationship, network pharmacology and components absorbed in plasma could be an effective way for rapid analysis and discovery of Q-marker in Shenzhiling Oral Liquid, which will be of great significance both to improve the quality control and evaluation, and ensure the safety and effectiveness of traditional Chinese medicines.
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Objective@#To develop a method of quantitative analysis of multi-components by single marker for nine effective constituents in Chenxiang-Huaqi tablets.@*Methods@#Using HPLC method, the mobile phase was acetonitrile -0.2% physcion acid in gradient elution program, the flow rate was 1.0 ml/min. The detection wavelength was set at 230 nm for liquiritin, ammonium glycyrrhizinate, costunolide and dehydrocostus lactone, and 283 nm for narirutin, naringin, hesperidin and neohesperidin, and 310 nm for pogostone. The column temperature was 30 ℃ and the injection volume was 10 μl. Using Hesperidin as the internal standard, the relative correction factors among liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were detected by QAMS and their contents were calculated. The content difference of the above nine components with the external standard method were compared.@*Results@#The calculated values of hesperidin, liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were not statistical significance from those measured by external standard method (P>0.05).@*Conclusions@#The method offered reference for quality standard of Chenxiang-Huaqi tablets.
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Objective To develop a method of quantitative analysis of multi-components by single marker for nine effective constituents in Chenxiang-Huaqi tablets. Methods Using HPLC method, the mobile phase was acetonitrile -0.2% physcion acid in gradient elution program, the flow rate was 1.0 ml/min. The detection wavelength was set at 230 nm for liquiritin, ammonium glycyrrhizinate, costunolide and dehydrocostus lactone, and 283 nm for narirutin, naringin, hesperidin and neohesperidin, and 310 nm for pogostone. The column temperature was 30 and the injection volume was 10 μl.℃ Using Hesperidin as the internal standard, the relative correction factors among liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were detected by QAMS and their contents were calculated. The content difference of the above nine components with the external standard method were compared. Results The calculated values of hesperidin, liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were not statistical significance from those measured by external standard method (P>0.05). Conclusions The method offered reference for quality standard of Chenxiang-Huaqi tablets.
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Chinese pharmacopoeia stipulates that the content of liquiritin in licorice slices should be no less than 0.5%. However, there are lots of unqualified licorice slices in the herbal medicine markets. Due to the important role of functional gene polymorphism in secondary metabolism, this study attempts to analyze the influence of chalcone synthase (CHS) gene polymorphism on liquiritin biosynthesis and find out the unique haplotypes in licorice samples with high or low content of liquiritin, and to provide a basis for further analysis of molecular mechanism in flavonoid biosynthetic pathway. The contents of the 4 main flavonoids (liquiritin, isoliquiritin, liquiritigenin, isoliquiritigenin) in 60 licorice samples were assayed by HPLC and the results were analyzed by Spearman and χ2 tests. The contents of the 4 main flavonoids were related to each other and obviously different in different original plants. They were highest in Glycyrrhiza uralensis samples and lowest in Glycyrriza inflate samples. Five G. uralensis samples with the highest liquiritin contents and five G. inflate samples with the lowest liquiritin contents were selected to clone the CHS cDNA sequences. 336 CHS cDNA sequences with a full length of 1 175 bp were obtained, 249 variable sites (141 missense mutation sites) were found, and 137 haplotypes were determined. 130 variable sites were found in the 336 CHS amino acid sequences and 102 types were determined. AA-3 is the major type of CHS in licorice, AA-35 is the special major type of CHS in the group with high flavonoids contents and AA-36 is the special major type of CHS in the group with low flavonoids contents. The mutation sites between AA-35 and AA-36 are I/V at 193 and V/T at 229. Discovery Studio 2.5 analysis of the three-dimensional structure of the CHS protein shows that the valine at site 229 of AA-35 is combined with malonyl-CoA. Homology analysis indicates that the homology of CHS among different species is low. This study is significant for identification of the unique haplotypes in licorices with high or low content of liquiritin and guiding the further molecular breeding of high-quantity licorice.
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To preliminarily investigate the dissolution behavior of Fuzi Lizhong pill, provide the basis for its quality control and lay foundation for dissolution behavior by determining the dissolution rate of liquiritin and glycyrrhizic acid. High-performance liquid chromatography (HPLC) method for simultaneous content determination of the two active ingredients of liquiritin and glycyrrhizic acid in Fuzi Lizhong pill was established; The dissolution amount of these two active ingredients in fifteen batches of Fuzi Lizhong pill from five manufacturers was obtained at different time points, and then the cumulative dissolution rate was calculated and cumulative dissolution curve was drawn. The similarity of cumulative dissolution curve of different batches was evaluated based on the same factory, and the similarity of cumulative dissolution curve of different factories was evaluated based on the same active ingredients. The dissolution model of Fuzi Lizhong pill based on two kinds of active ingredients was established by fitting with the dissolution data. The best dissolution medium was 0.25% sodium lauryl sulfate. The dissolution behavior of liquiritin and glycyrrhizic acid in Fuzi Lizhong pill was basically the same and sustained release in 48 h. Three batches of the factories (factory 2, factory 3, factory 4 and factory 5) appeared to be similar in dissolution behavior, indicating similarity in dissolution behavior in most factories. Two of the three batches from factory 1 appeared to be not similar in dissolution behavior of liquiritin and glycyrrhizic acid. The dissolution data of the effective ingredients from different factories were same in fitting, and Weibull model was the best model in these batches. Fuzi Lizhong pill in 15 batches from 5 factories showed sustained release in 48 h, proving obviously slow releasing characteristics "pill is lenitive and keeps a long-time efficacy". The generally good dissolution behavior also suggested that quality of different batches from most factories was stable. The dissolution behavior of liquiritin and glycyrrhizic acid in different factories was different, suggesting that the source of medicinal materials and preparation technology parameters in five factories were different.
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With annual Glycyrrhiza uralensis seedlings as experimental material, using "3414" optimal regression design and applied fertilizer, through the sampling of G. uralensis at harvest, root fresh weight and content of active components were measured in Lanzhou, Bayan Nur city, Chifeng, Jiuquan. Combined with NPK content in soil, potted experiments were used to study the effects of different nitrogen and phosphorus ratios on the dry matter accumulation and accumulation of active components of G. uralensis. The results reported as follows: the optimum fertilizer treatment in Lanzhou, Bayan Nur city, Chifeng, Jiuquan was N₁P₂K₁,N₂P₂K₁,N₁P₁K₂ and N₂P₁K₂, respectively. The efforts of single fertilizer on the fresh root weight acted as parabolic type.There was no significant effect of fertilizer treatment on the accumulation of active components of G. uralensis. Furthermore, in terms of nitrogen and phosphorus, the type of fertilizers that restricted the growth of the region was the type of elements with lower content in the soil. The optimal fertilizer usage was in inverse proportion to content of elements in soil. When the content of phosphorus in soil was low, nitrogen fertilizer and potash fertilizer showed positive interaction with phosphorus fertilizer, whereas, they showed negative interaction.
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Objective To establish fingerprint of Jinsangqi Kangdu Dropping Pills by HPLC; To control the quality of the preparation. Methods Waters XSELECT CSH-C18 chromatographic column (4.6 mm × 150 mm, 5 μm) was used and eluted with acetonitrile - 0.1% phosphoric acid solution gradient at the flow rate of 1.0 mL/min. The detection wavelength was 260 nm with column temperature of 30 ℃. Using calycosin-7-O-β-D-glucopyranoside, rutin, liquiritin, hyperoside, quercetin and ammonium glycyrrhizinate as the object references, ten batches of Jinsangqi Kangdu Dropping Pills were tested and analyzed by similarity comparison.Results Fringerprint spectrum of Jinsangqi Kangdu Dropping Pills had 24 common peaks in total, and characteristic spectrums of Hypericum Perforatum, Mori Cortex, Astragali Radix and Glycyrrhizae Radix et Rhizoma had been found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can evaluate the Jinsangqi Kangdu Dropping Pills quality totality,which can provide references for improving the quality control of the preparation.
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Objective:To develop a method of quantitative analysis of multi-components by single marker (QAMS) for six active constituents in Xiaoyao pills. Methods:Using glycyrrhizic acid as the reference, an HPLC method with a Kromasil C18 column (250 mm ×4.6 mm,5 μm) was applied,the mobile phase was acetonitrile(A)-0.1% phosphoric acid solution(B) with gradient elution (0-10 min,5 % A;10-25 min,5 % →20 % A;20-70 min,15 % →60 % A and 40-60 min,60 % A). The detection wavelength was 280nm,the column temperature was 35℃ and the injection volume was 10 μl. The relative correction factors among the six index components were detected by QAMS. The contents of the six index components were determined by an external standard method and QAMS to compare the results obtained from the two different methods and verify the practicability and stability of QAMS. Results:The established QAMS was used to determine the six index components in Xiaoyao pills,and totally 6 batches of Xiaoyao pills were determined.There were no significant differences in the calculated values and the determined ones (P > 0.05). Conclusion:The QAMS method is simple,effective and accurate in determining the contents of the six index components in Xiaoyao pills,which can be used for the quality control of Xiaoyao pills and provide reference for the further study.
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Objective:To develop a method of quantitative analysis of multi-components by single marker(QAMS)for the determi-nation of five constituents(ephedrine hydrochloride,amygdalin,liquiritin, baicalin and ammonium glycyrrhizinate)in Xiao'er Magan granule. Methods:Amygdalin was used as the internal reference substance, and the relative correlation factors(RCF) of ephedrine hydrochloride,liquiritin,baicalin and ammonium glycyrrhizinate to amygdalin were calculated and evaluated. The contents of the five constituents were determined by the external standard method(ESM) and QAMS,respectively. The content results determined by the two methods were compared and the feasibility of QAMS method was verified. Results:The RCF between amygdalin and the other con-tents was 1.237,1.318,1.327 and 0.884,respectively. There were no significant differences in the results between QAMS and ESM with the relative errors less than 0.3%. Conclusion:The QAMS method is accurate and feasible for the simultaneous determination of Xiao'er Magan granule.
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Objective: To study the chemical constituents of Lianhua Qingwen Capsules. Methods: The compounds were isolated and purified by gel column chromatography, MPLC, and preparative HPLC from 50% ethanol fraction of macroporous resin column chromatography of Lianhua Qingwen crude extracts. Their structures were elucidated by the spectral analyses. Results: Eight compounds were isolated and identified as 10-O-(p-hydroxycinnamoyl)-adoxosidic acid (1), aloe-emodin-8-O-β-D-glucopyranoside (2), quercitrin (3), matairesinol-4’-O-β-D-glucoside (4), liquiritin apioside (5), epi-vogeloside (6), vogeloside (7), and caffeic acid ethyl ester (8). Conclusion: Compound 1 is a new compound named lianhua iridoid A. Compounds 5-8 are isolated from Lianhua Qingwen Capsules for the first time. This study provides substance foundation for chemical research of Lianhua Qingwen Capsules.