Single Standard To Determine Multi-components Research Articles

Differentiation Between Chaenomelis Fructus and its Common Adulterant, Guangpi Mugua.

The dried fruit of Chaenomeles speciosa, known as Chaenomelis Fructus or Zhoupi Mugua, is a type of traditional Chinese medicine (TCM) that is widely used to treat many diseases. Guangpi Mugua, the dried fruit of the Chaenomeles sinensis, is its most commonly known adulterant. To establish a robust approach for the quality control and identification of Chaenomelis Fructus. Thin-layer chromatography (TLC) was optimized and used to discriminate Chaenomelis Fructus from Guangpi Mugua. High-performance liquid chromatography (HPLC) combined with fingerprint analysis and partial least-squares (PLS) discrimination analysis (DA) was employed to study the chemical differences between Chaenomelis Fructus and Guangpi Mugua. The single standard to determine multi-components (SSDMC) method, with credible precision, repeatability, stability, and durability, was developed for quantitative analysis of the abundant markers. The developed TLC and HPLC methods were effective in the authentication of Chaenomelis Fructus. Moreover, oleanolic acid, ursolic acid, pomolic acid, corosolic acid, 3-O-acetylpomolic acid, and one unknown compound were identified to be critical markers for the discrimination of Chaenomelis Fructus from Guangpi Mugua. Adulteration has always been a challenge in the development of TCM. This study presents useful insights that may help solve the problem of adulteration during the preparation of Chaenomelis Fructus. The present study provides a systematic method for the quality control of Chaenomelis Fructus. This is therefore the first step towards solving the problem of adulteration to improve the clinical safety and effectiveness of Chaenomelis Fructus.

Characterization and determination of free phytosterols and phytosterol conjugates: The potential phytochemicals to classify different rice bran oil and rice bran

Phytosterols are important beneficial compounds found in rice bran (RB) and rice bran oil (RBO). Although relationships have been confirmed between the forms of phytosterols and their bioactivities, the analysis of different forms of phytosterols in RB and RBO has been lacking. In this study, high temperature gas chromatography-mass spectrometry (HTGC-MS) was combined with the single standard to determine multi-components (SSDMC) method to determine free sterols (FSs) and steryl glycosides (SGs) in RB and RBO. High-performance liquid chromatography (HPLC) was used to determine steryl ferulates (SFs). There was clear variation in the composition of FS, SF and SG, indicating that different forms of phytosterols can discriminate between different RB and RBO. The developed method may be also useful for the detection of other compounds of interest in oils, oil seeds or cereals.

Deeper Chemical Perceptions for Better Traditional Chinese Medicine Standards

Abstract Traditional Chinese medicines (TCMs), a complex system of natural resources with many diverse components, are widely used as approved medicinal agents in China. Quality control of TCMs is a huge challenge for the government and for testing institutes and is associated with numerous scientific issues. Among these considerations include the following questions: How many components are in TCMs? How can the multiple components in TCMs be comprehensively delineated and subsequently characterized? What is the level and range of these (active) metabolites within these multiple-component TCMs, in order to recommend standards? and What are the qualities required for a marker constituent to be selected, and from a practical perspective, how can these components be assessed with low cost and in a short time? All of these factors require significant and deep thinking in order to understand the individualistic chemistry of TCM in order to develop enhanced TCM quality standards for improved and consistent patient care. In this review, the latest exploratory research in TCM chemistry analytical techniques and methods is summarized in order to begin to develop responses to these scientific issues. Advances in these methods have included multidimensional separation for liquid chromatography–high-resolution mass spectrometry (LC–HRMS), smart triggering data-dependent acquisition of LC–HRMS, target analysis with liquid chromatography–mass spectrometry (LC–MS), supercritical fluid chromatography, and data mining of large mass spectrometry (MS) datasets. In addition, two quality strategies have been introduced in order to save reference standards and the analysis time for a TCM quality standard, including the application of the single standard to determine multi-components (SSDMC) and monomethod-heterotrait matrix methods. Finally, a series of future improvements for analytical methods for TCMs are proposed.

Simultaneous quantitative analysis of 11 flavonoid derivatives with a single marker in persimmon leaf extraction and evaluation of their myocardium protection activity.

An improved, reliable and comprehensive method for assessing the quality of the ethyl acetate extract from persimmon leaves (EAPL) and its commercial preparation, Naoxinqing (Brain and Heart Clear capsules), has been developed and validated. Based on HPLC-DAD-ESI-Q-TOF-MS analysis, myricetin-3-O-β-D-galactoside (1), myricetin-3-O-glucoside (2), quercetin-3-O-β-D-galactoside (3), quercetin-3-O-β-D-glucoside (4), quercetin-3-O-(2″-O-galloyl-β-D-galactoside) (5), quercetin-3-O-(2″-O-galloyl-β-D-glucoside) (6), kaempferol-3-O-β-D-galactoside (7), kaempferol-3-O-β-D-glucoside (8), kaempferol-3-O-(2″-O-galloyl-β-D-galactoside) (9), kaempferol-3-O-(2″-O-galloyl-β-D-glucoside) (10), quercetin (11) and kaempferol (12) were identified from 15 batch samples. A HPLC fingerprint analytical method was established. All compounds, with the exception of compound 2, were simultaneously quantified by the single standard to determine multi-components (SSDMC) method, using kaempferol-3-O-β-D-glucoside as the internal standard. The rate of analysis was found to be faster with the SSDMC method than with current acid hydrolysis method (Pharmacopoeia of the People's Republic of China 2015 edition) and the results were more intuitive and reliable. Three-dimensional principal component analysis revealed that there were similar characteristics in persimmon leaf from same district. Analysis of the myocardial cell protection activity of 11 monomeric compounds showed that compounds 12, 11 and 10 were the main active ingredients that produce pharmacologic functions in EAPL. Among these compounds, the bioactive constituent of myricetin-3-O-β-D-galactoside was determined for the first time in Diospyros khaki. Thus, we have established an effective assessment method that can be applied to the comprehensive quality evaluation of EAPL extract and Naoxinqing capsule.

Construction of an optimized method for quality evaluation and species discrimination of Coptidis Rhizoma by ion-pair high performance liquid chromatography combined with response surface methodology

Coptidis Rhizoma (CR), the dried rhizome of three perennial Coptis specices, was widely used as a famous herbal medicine in China. Although the quantification of main alkaloids in CR has been extensively conducted, the existing analytical methods suffer from some flaws that restrict the general applicability in the routine quality assessment. In this work, we constructed an optimized method for quality evaluation and species discrimination of CR by ion-pairing high performance liquid chromatography (IP-HPLC) combined with response surface methodology (RSM). By employing Box-Behnken designs (BBD), 30 sets of experimental runs were performed to build the response surface models, and Derringer’s desirability was used to optimize the IP-HPLC separation conditions by simultaneously taking resolutions between two pairs of hardly − separated alkaloids and the retention time of the last eluted analyte as optimization criteria. Meanwhile, a single standard to determine multi-components (SSDMC) method based on the optimized IP-HPLC was set up and fully validated, to simultaneously determine six alkaloids including jatrorrhizine (JAT), columbamine (COL), epiberberine (EPI), coptisine (COP), palmatine (PAL) and berberine (BER), using BER as internal standard. Finally, the quantitative data from 33 batches of CR samples were comparatively analyzed, and the ratios of JAT/COL and EPI/JAT were discovered for species classification. Collectively, the established IP-HPLC method can be adopted for comprehensive quality evaluation and species discrimination of CR due to its general applicability.

A single marker choice strategy in simultaneous characterization and quantification of multiple components by rapid resolution liquid chromatography coupled with triple quadrupole tandem mass spectrometry (RRLC-QqQ-MS)

Single standard to determine multi-components (SSDMC) method has been accepted as an efficient technique for the quality control of Traditional Chinese medicines (TCMs), especially for overcoming the shortage of reference standards. HPLC-UV methods have been applied to establish SSDMC method for quantitative analysis in several plant medicines and Chinese patent medicines, however, no LC–MS methods have been used. The purpose of this study is to put forward an improved strategy for the choice of single marker in SSDMC using rapid resolution liquid chromatography coupled with triple quadrupole tandem mass spectrometry (RRLC-QqQ-MS). Five different Panax genus plants, recorded in the Chinese Pharmacopeia 2015 edition, were used as research subjects. An improved SSDMC strategy for simultaneous characterization and determination of 18 bioactive saponins in five Panax plants was put forward, and which was validated to be more superior. Then, it was fully investigated with respect to linearity, LODs, LOQs, precision and accuracy. Coupling with multivariate statistical analysis, the established and validated SSDMC strategy could be successively used in discrimination of the five Panax genus plants.

Comparison of five Lonicera flowers by simultaneous determination of multi-components with single reference standard method and principal component analysis

The flowers of Lonicera japonica Thunb. were extensively used to treat many diseases. As the demands for L. japonica increased, some related Lonicera plants were often confused or misused. Caffeoylquinic acids were always regarded as chemical markers in the quality control of L. japonica, but they could be found in all Lonicera species. Thus, a simple and reliable method for the evaluation of different Lonicera flowers is necessary to be established. In this work a method based on single standard to determine multi-components (SSDMC) combined with principal component analysis (PCA) for control and distinguish of Lonicera species flowers have been developed. Six components including three caffeoylquinic acids and three iridoid glycosides were assayed simultaneously using chlorogenic acid as the reference standard. The credibility and feasibility of the SSDMC method were carefully validated and the results demonstrated that there were no remarkable differences compared with external standard method. Finally, a total of fifty-one batches covering five Lonicera species were analyzed and PCA was successfully applied to distinguish the Lonicera species. This strategy simplifies the processes in the quality control of multiple-componential herbal medicine which effectively adapted for improving the quality control of those herbs belonging to closely related species.

Design of ultraviolet wavelength and standard solution concentrations in relative response factors for simultaneous determination of multi-components with single reference standard in herbal medicines

Single standard to determine multi-components (SSDMC) is a practical pattern for quality evaluation of herbal medicines (HMs). However, it remains challenging because of potential inconsistency of relative response factors (RRF) on different instruments. In this work, the effects of two key roles, i.e., ultraviolet (UV) wavelength and standard solution concentrations, on reproducibility of RRF were investigated. The effect of UV wavelength on reproducibility of RRF was studied by plotting the relationship of the peak area ratios (internal standard vs analyte) to wavelengths. The preferable wavelength should be set at the flat parts of the curve. Optimized 300nm produced a 0.38% RSD for emodin/emodin-8-O-β-D-glucopyranoside on five instruments, much lower than 2.80% obtained from the maximum wavelength at 290nm. Next, the effects of standard solution concentrations of emodin on its response factor (RF) were investigated. For one single point method, low concentration less than 49b/k resulted in significant variations in RF. For emodin, when the concentration is higher than 7.00μgmL−1, a low standard deviation (SD) value at 0.13 was obtained, while lower than 7.00μgmL−1, a high SD at 3.71 was obtained. The developed SSDMC method was then applied to determination of target components in 10 Polygonum cuspidatum samples and showed comparable accuracy to conventional calibration methods with deviation less than 1%.

A Comprehensive Method for Quality Evaluation of Houttuyniae Herba by a Single Standard to Determine Multi-components, Fingerprint and HPTLC Method.

A method of quality evaluation by a single standard to determine multi-components (SSDMC), fingerprint and high-performance thin layer-chromatography (HPTLC) was developed for traditional Chinese medicine (TCM), and validated with dry Houttuyniae Herba (HH). In quantitative analysis, an SSDMC method involving nine components has been established with the desirable linearity (r(2) ≥ 0.9998), precision (RSD < 2.7%), accuracy (97.4 - 103.1%) and ruggedness. Compared with the results obtained using the external standard method (ESM), this alternative SSDMC method was found to have no statistically significant differences. In fingerprint analysis, nine of fourteen peaks were identified. Simultaneously, 15 HH samples from different origins were classified by similarity analysis and hierarchical clustering analysis (HCA). Additionally, the HPTLC method with three flavonoid markers was firstly established. The combination of SSDMC with the fingerprint and HPTLC method has been verified in the quality control of HH both quantitatively and qualitatively, and will provide a new quality evaluation pattern for TCMs.

Strategies for elaboration of comprehensive quality standard system on traditional Chinese medicine

Based on the research goal of "traditional Chinese medicine standards lead in the international standard-setting", scientific strategies for the elaboration of traditional Chinese medicine (TCM) comprehensive quality standard system were introduced. TCM is a complex multi-component system which was used under the guidance of traditional Chinese medical theory. The present paper has put forward the basic principle of " deep research and simplified standard" to construct the quality standards of TCM. " Deep research" refers to systematic, thorough investigations on active constituents and biological mechanisms. On the basis of deep research, "simplified standard" means to establish a scientific and feasible standard, which should be practical and less complicated, to control the quality of Chinese herbs. Three key issues related to the elaboration of the quality standards are suggested. For reference substances, we propose the research methodology using reference extract to develop the qualitative and quantitative determination methods. Compared with that of using reference compounds, the preparation of reference extract is more accessible, economical and practical. Moreover, compared with reference crude drugs, the reference extract showed better batch-to-batch consistency. For identification, in addition to conventional methods, high performance liquid chromatography (HPLC), chemical fingerprints and characteristic chromatogram are proposed, in which more major marker compounds are monitored. And then LC-MS technique is employed to comprehensively analyze and characterize the peaks in the fingerprint. For multi-component quantification, the method of single standard to determine multi-components (SSDMC) is suggested, providing solutions for the lack of reference standards in quality evaluation. The SSDMC method uses a single reference standard to simultaneously determine the content of multiple compounds. In general, the integrate quality control standard of TCM is established based on combining innovative technology with practical applications.

A Novel Strategy using Single Standards to Determine Multi-components in order to Quantify the Major Macrocyclin Diterpenoids and Triterpenes in the Roots of Euphobia kansui

It was hardly practical to control the quality of Chinese herbal medicines with different types of constituents, broad polarities, and a variety of UV absorptions utilizing a single standard to determine multi-components (SSDMC) method alone. To extend the application range of the SSDMC method, a new strategy was developed and named as WP-ERS-CSP-SSDMC. This new strategy combined three different techniques, namely wavelength program (WP), exogenous reference standard (ERS) and core-shell particle (CSP), to enhance the existing SSDMC method. Taking Kansui Radix (roots of Euphobia kansui) as an example, thirteen major components including three jatrophane diterpenoids, eight ingenane diterpenoids and two triterpenes which included three pairs of isomerides and three strong hydrophobic components were all simultaneously assayed using this new strategy with ethyl benzoate, a synthetic compound, employed as a single reference standard for the first time. A C8 column packed with 2.7 µm core-shell particles was optimized to separate these constituents in 25 minutes on a conventional HPLC instrument with a photodiode array detector on which a four-stage wavelength program was established. The WP-ERS-CSP-SSDMC method was validated to have the capacity to acquire more repeatable and consistent results than that obtained by traditional external standard methods. Through Plackett-Burmantest (PB) tests, eight primary chromatographic parameters of the method were investigated and it was found that the two factors, wavelength and flow rate, should be strictly restrained. At last 75 batches of Kansui Radix and its three different processing products were successfully analyzed and discriminated by applying the proposed method. This work provided a useful strategy for SSDMC method making the simultaneous multi-component assay of complex TCM system achievable, and an extensive application of multi-component quality control of TCM feasible.

Ruggedness and robustness of conversion factors in method of simultaneous determination of multi-components with single reference standard

Single standard to determine multi-components (SSDMC) is a novel and rational method for quality control of botanical products and traditional Chinese medicines (TCMs). However, it is restricted to wide application due to unknown fluctuation in conversion factors when it is performed in different laboratories. To evaluate the fluctuations of conversion factors, we selected Salvia miltiorrhiza as an example to determine three components of tanshinones by SSDMC method. Then ruggedness and robustness test were adopted to comprehensively investigate three kinds of factors that may influence stability of conversion factors, which were related with environmental parametric variables, operational parametric variables and peak measurement parametric variables. Nested-factorial-design was used to perform ruggedness tests. One-variable-at-a-time (OVAT) procedure and Plackett–Burman (PB) design were both used in robustness test. The results showed that stability of conversion factors was principally related with accuracy of wavelength of UV detector, peak measurement parameters and concentration of standard solution. The acceptable range of conversion factors was obtained from robustness test. Our results showed that conversion factors were inevitable to change, but when key parameters were well controlled, the range of its fluctuation was acceptable and the SSDMC method could be used widely in different laboratories.