Quantitative Analysis Of Multi-components By Single-marker Research Articles

Determination of 13 chemical components of Epimedii Folium in pharmacopoeia by UPLC method combined with quantitative analysis of multicomponents by single-marker

The quantitative analysis of multicomponents by single-marker(QAMS) was established for 13 chemical components of Epimedii Folium, including neoglycolic acid, chlorogenic acid, cryo-chlorogenic acid, magnolidine, hypericin, epimedin A, epimedin B, epimedin C, icariin, baohuoside Ⅱ, sagittatoside A, icariin subside Ⅰ, and baohuoside Ⅰ, so as to investigate the feasibility and accuracy of this method in evaluating the quality of Epimedii Folium materials from different origins and different varieties. Through the scientific and accurate investigation of the experimental method, the external standard method was used to determine the content of 13 chemical components in epimedium brevieornu. At the same time, icariin was used as the internal standard, and the relative correction factors of icariin with neoglycolic acid, chlorogenic acid, cryo-chlorogenic acid, magnolidine, hypericin, epimedin A, epimedin B, epimedin C, icariin, baohuoside Ⅱ, sagittatoside A, icariin subside Ⅰ, and baohuoside Ⅰ were established, respectively. The contens of neoglycolic acid, chlorogenic acid, cryo-chlorogenic acid, magnolidine, hypericin, epimedin A, epimedin B, epimedin C, icariin, baohuoside Ⅱ, sagittatoside A, icariin subside Ⅰ, and baohuosideⅠ in Epimedii Folium were calculated by QAMS. Finally, the difference between the measured value and the calculated value was compared to verify the accuracy and scientific nature of QAMS in the determination. The relative correction factor of each component had better repeatability, and there was no significant difference between the results of the external standard method and those of QAMS. With icariin as the internal standard, QAMS simultaneously determining neoglycolic acid, chlorogenic acid, cryo-chlorogenic acid, magnolidine, hypericin, epimedin A, epimedin B, epimedin C, icariin, baohuoside Ⅱ, sagittatoside A, icariin subside Ⅰ, and baohuoside Ⅰ can be used for quantitative analysis of Epimedii Folium.

Simultaneous determination of glucosinolates and flavonoids in leaves of Moringa oleifera by quantitative analysis of multi-components by single-marker (QAMS)

To establish a method for simultaneously determining the content of four glucosinolates and five flavonoids in leaves of Moringa oleifera via quantitative analysis of multi-components by single-marker(QAMS) and verify the feasibility and applicability of the established method. The glucosinolates and flavonoids were analyzed via Waters Acquity UPLC HSS T_3 column(2.1 mm × 100 mm, 1.8 μm). The gradient elution was carried out with the mobile phase composed of 0.1% formic acid-acetonitrile and 0.3% formic acid at the flow rate of 0.4 mL·min~(-1) and the column temperature of 40 ℃. The wavelengths for the detection of glucosinolates and flavonoids were 225 nm and 260 nm, respectively. With 4-O-(α-L-rhamnoyloxy)-benzyl glucosinolate and vecenin-2 as internal reference substances, the relative correction factors of four glucosinolates and five flavonoids were respectively calculated for determining the content of the 9 ingredients in leaves of M. oleifera. To verify the accuracy and feasibility of QAMS, we used internal standard method(ISM) and external standard method(ESM) to determine the content of glucosinolates and flavonoids, respectively. The t-test results showed that there was no significant difference in the content of glucosinolates obtained by ISM and QAMS methods or in the content of flavonoids obtained by ESM and QAMS methods. The content of glucosinolates and flavonoids varied among M. oleifera of different varieties and from different producing areas. The total glucosinolates and total flavonoids had the highest content in the Indian variety while the lowest content in the variety ■Honghe No. 1'. The established QAMS method is rapid, simple and accurate and can be used for simultaneous determination of glucosinolates and flavonoids in the leaves of M. oleifera. This study provides experimental data for the quality control and utilization of M. oleifera leaves.

A trinity fingerprint evaluation system of traditional Chinese medicine

This study focused on developing a set of quality evaluation methods that can reflect the multi-levels and multi-characteristics of traditional Chinese medicine (TCM). Taking licorice (Glycyrrhiza glabra L.) as the method development sample, the feasibility of multi-markers assay by monolinear method (MAML) was explored through the standard curve relationship among active components for the first time. Using glycyrrhizic acid as measurement marker, MAML can simultaneously quantify five components of licorice, including isoliquiritigenin, isoliquiritin apioside, liquiritigenin, liquiritin and liquiritin apioside. Comparing MAML and quantitative analysis of multi-components by single-marker (QAMS) to the external standard method (ESM) respectively, it was found that there was no significant difference in the content of components that were calculated by MAML and ESM (the relative error (RE) was generally less than 2.00%). However, the RE of the component content calculated by QAMS fluctuated greatly, indicating that the MAML was more accurate than QAMS. In addition, UV and THz quantum fingerprints were initiated by the interval erasure method. Taking the systematically quantified fingerprint method as the core, a "Trinity" fingerprint quality evaluation system based on HPLC, UV and THz was developed. The system successfully distinguished the quality differences of licorice samples from 13 producing areas and two ecological models by the comprehensive evaluation results. Simultaneously, the quality information of licorice at different technical levels was revealed. Finally, bivariate correlation analysis was used to examine the linkage between UV/HPLC and antioxidant spectrum efficacy, and the two-dimensional activity spectrum of licorice was provided. It may furnish a more thorough and objective analytical technique for licorice and even other TCMs in chemical fingerprint features, chemical bond vibration characteristics and biological activity information.

Quantitative analysis of multi-components by single-marker: An effective method for the chemical characterization of wood vinegar

Wood vinegar is a complex mixture of organic compounds. QAMS is a relatively simple and cost-effective method which is used widely in recent years. In this study, quantitative analysis of multi-components by single-marker (QAMS) was applied to quantify the main volatile chemical components in wood vinegar for the first time. Using acetic acid as an internal reference compound, the relative correction factors of a further 19 chemical volatile components were obtained. 20 wood vinegar prepared from different lignocellulosic biomass, and at different pyrolysis temperatures, were analyzed by QAMS and the data compared with those obtained using an internal standard (phenethyl acetate) method. The results showed that the precision and repeatability of the QAMS method were good, and the responses obtained for each compound using GC/MS were linear over the corresponding concentration range (R2 ≥ 0.999). The average spike recoveries of 20 volatile compounds from wood vinegar ranged from 92.65% to 105.93% (RSD ＜ 7%, n = 6). There were no significant differences (P > 0.05) in the results obtained from the analysis of three batches of wood vinegar samples (RSD < 2%) using both methods. The volatile composition of samples obtained from wood vinegar prepared from the pyrolysis of different materials and at different temperatures were significantly different. The successful application of QAMS to the quantitative analysis of the composition of wood vinegar not only may provide a theoretical reference for further developments in the production, pyrolysis conditions, and modes of action for wood vinegar, but also provides ideas for the application of QAMS in other fields.

Application of QAMS for quality evaluation and control of Chinese patent medicines:taking Bufonis Venenum-contained preparations as examples

As a new strategy capable of uncovering the characteristics of traditional Chinese medicines, the quantitative analysis of multi-components by single-marker(QAMS) has been widely employed for the quality evaluation of Chinese medicinal materials, slices, and extracts. However, its application in the assessment of Chinese patent medicines is yet to be explored. By referring to the determination of three bufogenins in Bufonis Venenum by QAMS described in Chinese Pharmacopoeia(2020 Edition), this paper selected seven representative preparations containing Bufonis Venenum and explored whether the relative correction factors(RCFs) of cinobufagin(CB) to bufalin(BF) and resibufogenin(RB) could be directly used for the quality control of Bufonis Venenum-contained preparations. Based on the qualitative analyses under the same chromatographic conditions as used for toad venom, combing specificity test, five preparations such as Yatong Yili Pills, Houzheng Pills, Xiongdan Jiuxin Pills, Liushen Pills and Niuhuang Xiaoyan Pills, were expected to use validated RCFs for the direct determination of three components. Taking Houzheng Pills as an example, the methodological validation of bufalin, cinobufagin and resibufogenin was carried out, and the recoveries of bufalin, cinobufagin and resibufogenin were 90.64%-106.1%. The obvious difference was not observed between the contents of bufalin and resibufogenin in 24 batches of preparation samples by QAMS and external reference method. In the tested samples, the content of bufalin, cinobufagin and resibufogenin were 1.27-2.61, 2.44-5.66 and 0.988-3.16 mg·g~(-1) in 10 batches of Liushen Pills samples. The contents of bufalin, cinobufagin and resibufogenin were 0.760-1.32, 1.35-2.39 and 0.600-1.55 mg·g~(-1) in 10 batches of Houzheng Pills samples from three manufacturers. The obtained data contribute to improving the quality standard of Bufonis Venenum-contained preparations, and they also provide some ideas for the application of QAMS in the quality evaluation and control of Chinese patent medicines.

Simultaneous Determination of Eight Chemical Components in Angelicae Sinensis Radix and Its Herbal Products by QAMS.

A HPLC method has been developed for simultaneously detecting chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, coniferyl ferulate, senkyunolide A, ligustilide, and levistolide A in Angelicae Sinensis Radix through quantitative analysis of multicomponents by single-marker (QAMS) method with ferulic acid as internal standard substance. The relative analysis correction factors of each component in Angelicae Sinensis Radix have good reproducibility under different chromatography conditions. In addition, no significant difference of results was found between quantitative analysis of multicomponents by single-marker (QAMS) method and external standard method in determining content of these components of different Angelicae Sinensis Radix and its 12 kinds of preparations. As a result, the established QAMS method for Angelicae Sinensis Radix analysis with ferulic acid as internal standard substance is accurate and feasible, which could be used as an effective and economical method to control quality of Angelicae Sinensis Radix and its herbal products.

New Determination Method of Two Bioactive Saponins in Lysimachia capillipes Hemsl. by Quantitative Analysis of Multi-Components by Single-Marker

Capilliposide B (CPS-B) and Capilliposide C (CPS-C), as the key components in Lysimachia capillipes Hemsl., increasingly aroused the interest and research concern of many researchers due to the good bioactivities. Nowadays, the reference standards of CPS-B and CPS-C yield were very limited. Due to the deficit of reference standards, the determination could be difficult to carry out, and the quality control and evaluation would be restrained afterwards. To solve this urgent problem, a quantitative analysis of multi-components by single-marker (QAMS) method was proposed and established based on high-performance liquid-chromatography tandem evaporative light-scattering detector. In this QAMS method, the content of the two bioactive components could be calculated by buddlejasaponin IV, which is applied as an external standard and readily obtained. And the methodological experiments were evaluated and indicated accuracy, stability and feasibility of this QAMS method. Therefore, in this study, this built method would properly meet the requirement of determination of CPS-B, CPS-C and quality control of the L. capillipes Hemsl. plant.

Exploitation of HPLC Analytical Method for Simultaneous Determination of Six Principal Unsaturated Fatty Acids in Oviductus Ranae Based on Quantitative Analysis of Multi-Components by Single-Marker (QAMS).

As one of the featured products in northeast China, Oviductus Ranae has been widely used as a nutritious food, which contains a variety of bioactive unsaturated fatty acids (UFAs). It is necessary to establish a scientific and reliable determination method of UFA contents in Oviductus Ranae. In this work, six principal UFAs in Oviductus Ranae, namely eicosapentaenoic acid (EPA), linolenic acid (ALA), docosahexaenoic acid (DHA), arachidonic acid (ARA), linoleic acid (LA) and oleic acid (OA), were identified using UPLC-MS/MS. The UFAs identified in Oviductus Ranae were further separated based on the optimized RP-HPLC conditions. Quantitative analysis of multi-components by single-marker (QAMS) method was implemented in content determination of EPA, ALA, DHA, ARA and OA, where LA was used as the internal standard. The experiments based on Taguchi design verified the robustness of the QAMS method on different HPLC instruments and chromatographic columns. The QAMS and external standard method (ESM) were used to calculate the UFA content of 15 batches of Oviductus Ranae samples from different regions. The relative error (r < 0.73%) and cosine coefficient showed that the two methods obtained similar contents, and the method validations met the requirements. The results showed that QAMS can comprehensively and effectively control the quality of UFAs in Oviductus Ranae which provides new ideas and solutions for studying the active components in Oviductus Ranae.

Overall control herbal medicine in best consistency

The standard preparation (SP) based on the quantitative fingerprint was proposed to control the quality of compound licorice tablets (CPLTs) in this paper, which is a great way to control the quality consistency nowadays. Here, taking China 145 batches of CPLTs from 9 manufacturers as samples to set up High Performance Liquid Chromatography fingerprints and CPLT-SP, and employing the systematically quantified fingerprint method (SQFM) combined five markers’ contents to evaluate their quality. The results showed that all samples were qualified. The quantitative analysis of multi-components by single-marker (QAMS) for the determination of five components in CPLTs was applied, and there was no striking diversity in the results compared to the standard curve method. Furthermore, the correlation between raw materials and preparation of CPLTs was studied for predicting quality intelligently. As well as the ultraviolet full fingerprint dissolution was applied to evaluate the rationality of preparation. The results demonstrated that SQFM based on SP can integrally and perfectly control the quality of HD in best consistency.

Recommendations on quality standards of Andrographis Herba and its processed slices in Chinese Pharmacopoeia (2020 edition)

To improve and perfect the quality standards and propose recommendations for the revision of quality standards for Andrographis Herba and its processed slices in Chinese Pharmacopoeia(ChP)(2020 edition) based on the problems and limitations in ChP(2015 edition). TLC identification method with andrographolide and control herbs as references was established using silica gel G thin layer plate, with chloroform-methylbenzene-methanol(8∶1∶1) as developing solvent, and 10% sulfuric acid ethanol solution as colour-developing agent. This method has good reproducibility, strong specificity and high sensitivity. As compared with the original method in ChP 2015, this method has better development effect and clearer spots. Based on the previous research, a quantitative analysis of multi-components by single-marker(QAMS) method with andrographolide as the internal reference substance was developed to simultaneously determine the contents of 4 diterpene lactones: andrographolide(S), neoandrographolide(A), 14-deoxyandrographolide(B), and dehydroandrographolide(C). The relative correction factors of f_(A/S), f_(B/S), and f_(C/S) were determined as 1.12, 0.79, and 0.63, respectively. The relative retention time of t_(A/S), t_(B/S), and t_(C/S) was 1.95, 2.18, and 2.25, respectively. According to the content determination results in 46 batches of crude drugs and 38 batches of processed slices, it was stipulated that the total contents of 4 diterpene lactones should not be less than 1.5% and 1.2% in crude drugs and processed slices, respectively. As compared with the original method in ChP 2015, the present QAMS method could not only reduce the detection cost and improve the efficiency, but also can be used to evaluate the quality of Andrographis Herba and its processed slices more comprehensively and objectively. Diterpene lactones are generally recognized as the effective components in Andrographis Herba, and their contents in leaves were much higher than those in stems. However, almost all of the current commercial processed slices are processed from stems, so their quality is gene-rally poor and the efficacy is hard to be guaranteed. Therefore, the weight percentage of leaves should be added into the inspection items of the processed slices and it should not be less than 25%.

Effects of different harvesting and processing methods on quality of Andrographis Herba

The effects of the growing periods, processing methods, nutritive organs and cut parts of the herbs on the quality of Andrographis Herba have been studied, which could provide scientific basis for producing high-quality crude materials of Andrographis Herba. A quantitative analysis of multi-components by single-marker(QAMS) method was used to simultaneously determine 4 diterpene lactones of andrographolide(AD), neoandrographolide(NAD), 14-deoxyandrographolide(14-DAD), and dehydroandrographolide(DHAD) in 78 batches of Andrographis Herba samples. Visual analysis, clustering analysis and partial least squares discrimination analysis(PLS-DA) were used for data processing and mining. The growing periods of Andrographis Herba were divide into the rapid growing stage(Ⅰ), the luxuriant foliage stage(Ⅱ), the bud stage(Ⅲ), the flower stage(Ⅳ), the fruiting stage(Ⅴ) and the later fruiting stage(Ⅵ), six ones in total. The results showed that the total contents of 4 diterpene lactones(TC) kept almost constant in growing periods of Ⅰ-Ⅲ, began to fall off in growing period Ⅳ, decreased rapidly in growing period Ⅴ and reduced to minimum in growing period Ⅵ. Therefore, the best harvesting period for Andrographis Herba was the bud stage. For the processing methods, none significant difference occurred for TC between drying by the sun and drying in the shade. However, in comparison of the samples dried by the sun, the samples dried in the shade exhibited lower content of AD, nonetheless, the higher contents of DHAD and NAD, indicating that the process of drying in the shade may lead to the decomposition and transformation of AD. Therefore, when the weather conditions permitted, the method drying by the sun should be used. The distributions of AD, DHAD and TC in the different cut parts and nutritive organs were determined as follows: upper cut parts > middle cut parts ≈ entire herbs > lower cut parts, leaves > entire herbs > stems. Furthermore, the contents of 4 diterpene lactones in leaves and herbs were much higher than those in stems. Therefore, it was suggested that the upper and middle cut parts of the herbs or the parts containing leaves should be preferred for harvesting to ensure the proportion of the leaves in Andrographis Herba.

Establishment of QAMS method with dehydroandrographolide as internal reference substance for quality control of Andrographis Herba

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.

A quantitative method for simultaneous assay of seven active ingredients with one marker in Scrophularia ningpoensis root

To establish a new quantitative method for simultaneous determination of multi-components in Scrophularia root by using high performance liquid chromatography (HPLC) and validate its feasibilities．Meanwhile,using catalpol as one chemical reference substance to establish the relative correct factors and relative retention values of aucubin,harpagide,acteoside,angoroside C,harpagoside and cinnamic acid．Then using the quantitative analysis of multi-components by single-marker (QAMS)model,the six analytes can be quantitatively determined in Scrophularia root．The method was evaluated by comparison of the quantitative results to external standard method．No significant differences were observed between the quantitative results of the two methods．The obtained RCFs were credible．It is feasible and suitable to evaluate the quality of Scrophularia root.

Study on Quality Standard of Processed Curcuma Longa Radix.

To control the quality of Curcuma Longa Radix by establishing quality standards, this paper increased the contents of extract and volatile oil determination. Meanwhile, the curcumin was selected as the internal marker, and the relative correlation factors (RCFs) of demethoxycurcumin and bisdemethoxycurcumin were established by high performance liquid chromatography (HPLC). The contents of multicomponents were calculated based on their RCFs. The rationality and feasibility of the methods were evaluated by comparison of the quantitative results between external standard method (ESM) and quantitative analysis of multicomponents by single-marker (QAMS). Ethanol extracts ranged from 9.749 to 15.644% and the mean value was 13.473%. The volatile oil ranged from 0.45 to 0.90 mL/100 g and the mean value was 0.66 mL/100 g. This method was accurate and feasible and could provide a reference for further comprehensive and effective control of the quality standard of Curcuma Longa Radix and its processed products.

Integrative research of QAMS model used in assay of flavonoids in Epimedii Folium, its processed products and finished Chinese medicines

Through a series of methodology investigations, we established a new method for simultaneous analysis of epimedins A, B, C, icariin and baohuoside I in Epimedii Folium by using high performance liquid chromatography (HPLC). Meanwhile, using Icariin as an internal reference substance to establish the relative correct factors and relative retention values of Epimedins A, B, C and Baohuoside I to Icariin, then using the quantitative analysis of multi-components by single-marker (QAMS) model, the five analytes can be quantitatively determined in Epimedii Folium and its processed products as well as Kanggu Zengsheng capsule only by measuring the content of icariin in the corresponding samples. All these analysis are completed in the same chromatorgraphic conditions. This paper played the part of demonstration role in the popularization and application of QAMS method established in a single herb to the proprietary Chinese medicines containing this herb.

Quality Evaluation of Polar and Active Components in Crude and Processed Fructus Corni by Quantitative Analysis of Multicomponents with Single Marker.

Objective. To develop a quantitative analysis of multicomponents by single-marker (QAMS) method for the simultaneous determination of polar active components in Fructus Corni. Methods. Loganin was selected as the internal reference, and the relative correction factors (RCFs) of gallic acid, 5-hydroxymethyl-2-furfural, morroniside, sweroside, cornin, 7α-O-methylmorroniside, 7β-O-methylmorroniside, 7α-O-ethylmorroniside, 7β-O-ethylmorroniside, and cornuside were established. The contents of multicomponents were then calculated based on their RCFs, respectively. Contents of the 11 components were also calculated by external standard method and compared with those of the QAMS method. Results. The contents of the 11 components in 21 crude and 10 processed Fructus Corni products were measured. No significant difference was found in the quantitative results of the QAMS and external standard methods. Conclusion. QAMS could serve as an accurate and convenient method in determining the polar and active components in Fructus Corni and its processed products.

Establishment and validation of quantitative analysis of multi-components by single-marker for quality assessment of compound danshen preparations

Compound danshen preparations (CDPs) are used clinically for the treatment of cardiovascular and cerebrovascular diseases. By using the quantitative analysis of multi-components by single-marker (QAMS) method, sixteen compounds (danshensu, protocatechuic acid, protocatechuicaldehyde, caffeic acid, rosmarinic acid, lithospermic acid, notoginsenoside R1, salvianolic acid B, ginsenoside Rg1, ginsenoside Re, salvianolic acid A, salvianolic acid C, ginsenoside Rb1, ginsenoside Rd, cryptotanshinone, and tanshinone IIA were quantified on an ACQUITY ultraperformance liquid chromatography (UPLC) HSS T3 column (2.1 × 100 mm, 1.8 μm) with the mobile phase consisting of 0.1% formic acid aqueous solution (A) and acetonitrile (B) using a gradient elution at the flow rate of 0.30 mL/min in 30 min at 30°C, which was also validated by UPLC-diode array detection (DAD) and UPLC-electrospray ionization multistage/mass spectrometry (ESI-MS/MS) for assuring the feasibility and accuracy. Tested by robustness experiment under slightly changeable conditions, the stability of relative correction factor (RCF) proved to be stable, with RSDs below 5.69%, except for notoginsenoside R1 with relative standard deviation (RSD) 7.83%. This reliable and convenient QAMS method resolved the problem of standard substance insufficiency and improved the quality assessment of preparations consisting of complex compounds with different chemical structures, such as CDPs.

A simple and sensitive method for the simultaneous determination of six constituents in hawthorn leaves

The quantitative analysis of multi-components by single-marker (QAMS) method was developed and validated for the simultaneous determination of chlorogenic acid (a), vitexin-4′′-O-glucoside (s), vitexin-2′′-O-rhamnoside (b), vitexin (c), rutin (d) and hyperoside (e) in hawthorn leaves using high-performance liquid chromatography-diode array detection (HPLC-DAD), and then it was applied to the extract and its traditional Chinese herbal preparations for the actual production. In this study, the six constituents were first determined using the external standard (ES) method, and then vitexin-4′′-O-glucoside (s) was selected as the internal constituent to calculate the relative correction factors (RCF) between s and the other five constituents, respectively. We can calculate the content of the other five constituents with the RCF corresponding directly. Compared with the results obtained using the ES method, the QAMS is found to have no statistically significant difference in our laboratory as verified by the RSD 0.05 of paired T-test. The results indicate that QAMS is suitable for the simultaneous determination of multiple constituents when the reference substances are scarce, expensive or unstable. Moreover, it can reduce the cost of testing on the basis of improving the quality standard in the actual production.

Optimization of Scutellaria baicalensis extraction process by orthogonal experiment combined with pharmacodynamic index

To optimize the Scutellaria baicalensis extraction process, the filter paper method and the bacteriostatic ratio method were adopted to determine the in vitro bacteriostatic efficacy of water extracts and 60% alcohol extracts from S. baicalensis. The quantitative analysis of multi-components by single-marker (QAMS) was used to determined the contents of four active components, baicalin, wogonoside, baicalein and wogonin. In addition, with the bacteriostatic ratio and the overall desirability of the contents of four active components as indexes, the orthogonal experiment was adopted to detect the effect of water addition, extraction frequency and extraction time. The optimal extraction process was to add 12 times of water for the first time, 10 times of water for the second time, extract for 2 time, 2 h for each time. This optimization process is stable and feasible, with a higher bacteriostatic ratio in extracts.

Quality Evaluation of Astragali Radix Products by Quantitative Analysis of Multi-components by Single Marker

ObjectiveTo develop a quantitative analysis of multi-components by single-marker (QAMS) method for the simultaneous determination of eight components in Astragali Radix products, and to examine the feasibility of using the method among the different dosage forms and between two different types of compounds. MethodsEight main effective components, campanulin, genistin, ononin, calycosin, genistein, formononetin, methylnissolin, and astragaloside IV were selected as analytes for the quality control of Astragali Radix products. Calycosin was selected as the internal reference substance, the content of which was determined by external standard method; the relative correction factors (RCFs) of campanulin, genistin, ononin, genistein, formononetin, methylnissolin, and astragaloside IV were calculated. In total, twelve Astragali Radix specimen in decoction pieces, as well as in two different dosage forms, such as granule and oral liquid products, were used for the quality control by both methods of external standard and QAMS. The validity of the QAMS method was evaluated by comparison on the quantitative results of the two methods. ResultsThese RCFs were obtained with good reproducibility (RSD < 6.5%) by using ultra high performance liquid chromatography coupled with diode array detector under various chromatographic conditions. Meanwhile, no obvious differences (RSD < 3.98%) were found in the quantitative results of the seven components in twelve samples of Astragali Radix products determined by the two methods. ConclusionQAMS is a reliable and feasible method in determining the components in products of Astragali Radix.