Chemical Markers For Quality Control Research Articles

What Do We Know About CapsicumVolatilome?

The Capsicum genus includes several cultivated species that release complex blends of volatile organic compounds (VOCs) associated with their unique aroma. These VOCs are essential info-chemicals in ecological interactions. In this review, we describe how the volatilomic profiling naturally varies based on specific plant organs and genotypes as well as how non-beneficial organisms affect VOCs biosynthesis and accumulation in pepper plants. Also, we show evidence about VOCs variation under the pressure of different abiotic factors such as water stress, soil type and nutrient availability. The contribution of specific metabolic pathways and gene expression related to the biosynthesis of particular VOCs is addressed. We highlighted the utility of VOCs as chemical markers for quality control in the food industry, breeding programs to generate resistant plants and to improve aroma innovation. Herein we present a database containing 2734 VOCs, revealing 113 as the basic core of the volatilome from five Capsicum species.

Quantification and holistic quality evaluation of Wulingzhi extract by UHPLC-Q-Orbitrap-HRMS coupled with chemometric approaches.

The excreta of Trogopterus xanthipes ("Wulingzhi" in Chinese, WLZ) is a well-known traditional Chinese medicine. It has been used for centuries to treat amenorrhea, menstruation and postpartum abdominal pain. However, a systematic quality study on WLZ chemical markers has yet to be conducted. This study aimed to establish an ultra-high-performance liquid chromatography coupled with a hybrid quadruple extraction Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) method for the simultaneous quantitative determination of 20 compounds in 53 batches of WLZ; the method rapidly and sensitively determined the 20 plant- or animal-derived compounds. Firstly, the proposed approach was validated to satisfy the method's linearity, detection limits, precision, repeatability, stability and accuracy. Subsequently, multivariate analysis was used to identify correlations between the samples and feed, processing and regions. Finally, this method was used to further identify chemical markers for quality control in combination with chemometrics. This is the first report on pinusolide, betaine, hippuric acid, 4-oxorentinoic acid, 15-methoxypinusolidic acid and 4-oxoisotrentinoin in WLZ; the quality of WLZ became homogeneous after processing with vinegar (V-WLZ). Moreover, we screened for potential component markers, including uridine, allantoin, amentoflavone, hippuric acid, 3,4-dihydroxybenzoic acid, pinusolide, quercetin and kaempferol. These results were practical and efficient for the chemical clarification of WLZ and V-WLZ.

Comprehensive chemical profiling and quantitative analysis of ethnicYi medicine Miao-Fu-Zhi-Tong granules using UHPLC-MS/MS

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity. Miao-Fu-Zhi-Tong (MFZT) granules, an ethnic Yi herbal prescription, comprises 10 herbs and has been clinically applied for gouty arthritis (GA) therapy. Herein, a series of chemical profiling strategies including in-house library matching, molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were developed for qualitative analysis of MFZT granules. A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes. Moreover, network pharmacology analysis indicated that MFZT's anti-gout mechanism was mostly associated with the nuclear factor kappa-B (NF-κB) signaling, nucleotide oligomerization domain (NOD)-like signaling and rheumatoid arthritis pathways, along with the synergistic effect of 84 potential active compounds. In addition, a quantitative analytical method was developed to simultaneously determine the 29 potential effective components. Among them, berberine, pellodendrine, 3-feruloylquinic acid, neoastilbin, isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control. These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.

Integrating serum pharmacochemistry and network pharmacology to identify chemical markers for quality control of Apocyni Veneti Folium.

Apocyni Veneti Folium (AVF) is a commonly used traditional Chinese medicinal herb for the treatment of hypertension. Chemical markers are crucial for the quality control of herbal medicines; however, the therapeutic components of AVF remain to be well elucidated. This study was intended to integrate serum pharmacochemistry and network pharmacology to identify chemical markers of AVF and establish an efficacy-related quality control method of AVF. Ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was applied to identify the absorbed AVF constituents in rat serum. Network pharmacology was further used to identify anti-hypertension-related chemical markers. Subsequently, a quantitative method was established using UPLC with diode array detection (DAD) and applied for quality evaluation of commercial AVF samples. Thirteen prototype constituents were unequivocally or tentatively characterized in serum samples, among which quercetin, kaempferol, hyperoside, isoquercitrin, chlorogenic acid, cryptochlorogenic acid, and neochlorogenic acid were identified as dominant chemicals related to anti-hypertensive efficacy. The quantitative data showed that the total contents of seven marker components even showed 2-fold variation among 14 batches of commercial AVF samples with RSD values ranging from 12.15% to 75.61%. Hierarchical cluster analysis and heatmap analysis showed that 14 batches of commercial AVF samples could be divided into three main groups. The chemical markers obtained from this study could be applicable for efficacy-related quality control of AVF.

Quality Control of Oleum Cinnamomi Assisted by Network Pharmacology Strategy.

Oleum Cinnamomi is a traditional medicine used by the Hmong, the essential oil obtained from Fructus Cinnamomi, for the treatment of coronary heart disease. Information regarding the efficient quality control markers of it is lacking, which has become a bottleneck restricting its development and utilization. Here, an integrated qualitative analysis approach based on a GC-MS and network pharmacology strategy was applied to explore quality control markers for the assessment of Oleum Cinnamomi. Firstly, the compounds of Oleum Cinnamomi were detected by GC-MS. In total, 57 chemical components were identified, mainly monoterpenes and sesquiterpenes, accounting for 83.05% of total essential oil components. Secondly, network pharmacology was adopted to explore the compounds linked to target genes of coronary heart disease. Fifty-two compounds were found, indicating the effectiveness of Oleum Cinnamomi in the treatment of coronary heart disease. Among them, 10 compounds, including eucalyptol, were chosen as potential effective compounds in Oleum Cinnamomi. Thirdly, an established GC-MS SIM method was validated and applied for the simultaneous determination of the contents of these 10 compounds using 20 sample batches of Oleum Cinnamomi. It was preliminarily found that the contents of these 10 compounds differed in Oleum Cinnamomi from different origins. Finally, quantitative analyte data were analyzed using multivariate statistical analysis to determine Oleum Cinnamomi quality. Four compounds (eucalyptol, p-cymene, sabinene, β-pinene) were identified as chemical markers for quality control. Accordingly, this study provides new strategies to explore the quality control markers and develops a novel method for the quality assessment of Oleum Cinnamomi.

Chemical profiling and quantification of multiple components in Jin-Gu-Lian capsule using a multivariate data processing approach based on UHPLC-Orbitrap Exploris 240 MS and UHPLC-MS/MS.

The Jin-Gu-Lian capsule, a Chinese Miao herbal compound, is widely used to treat rheumatoid arthritis. In this study, a rapid, selective, and sensitive UHPLC-Orbitrap Exploris 240 MS method was developed to analyze the chemical composition of Jin-Gu-Lian capsules. A total of 88 compounds were identified, including 23 flavonoids, 23 organic acids, 14 phenylpropanoids, 12 phenols, eight alkaloids, four terpenes, three quinones, and one ketone. Among these, 21 compounds were clearly detected based on a comparison with reference standards and selected as quality control markers. Thereafter, these compounds were simultaneously determined in the Jin-Gu-Lian capsules. The established method was successfully validated and applied for the simultaneous determination of 21 biologically active compounds in Jin-Gu-Lian capsules of 27 sample batches. Quantitative data of the analytes were analyzed using multivariate statistical analysis to determine the quality of the Jin-Gu-Lian capsules. Four compounds (JGLC6 [salidroside], JGLC8 [chlorogenic acid], JGLC12 [liriodendrin], JGLC19 [quercetin]) were identified as chemical markers for quality control of Jin-Gu-Lian capsules. Altogether, the established method was validated as a novel and efficient tool, that can be used for rapid analysis of Jin-Gu-Lian capsules. Accordingly, this study serves as a reference for scientific research on traditional Chinese and ethnic medicine.

Comparative investigation on metabolites and biological activities of Paeonia ostii stamens from different geographical regions of China

Paeonia ostii is a widely cultivated medicinal and oilseed crop in China. The stamens of P. ostii are important by-products of this crop and conventionally used as tea with putative beneficial effects on human health. However, the in-depth understanding of P. ostii stamens is scarce. In this study, comparative analysis was carried out to systematically characterize the bioactive constituents and biological activities of P. ostii stamens from different geographical regions of China. Seventy-seven secondary metabolites were tentatively identified through untargeted metabolomic analysis based on MS/MS molecular similar networking and other approaches. Targeted metabolomics conjugated with principal component analysis revealed that eight bioactive compounds, including oxypaeoniflorin, pentagalloyl glucose (PGG), paeoniflorin, and albiflorin, were the primary chemical markers for quality control. Habitats substantially affected the content of PGG, and geographical factor of Nujiang (Southern China) significantly promoted the accumulation of PGG. The extracts from peony stamens across all geographical regions exhibited potent antioxidant, antibacterial, antitumor, and anti-neuroinflammation properties, which were strongly correlated with bioactive compounds. Our results provide new insights into P. ostii stamens to be used for pharmaceutical purposes.

Identification of C-glycosyl flavones and O-glycosyl flavones in five Dendrobium species by high-performance liquid chromatography coupled with electrospray ionization multi-stage tandem MS.

Flavones are widely used in traditional Chinese medicine (TCM) and are the pharmacologically active ingredients of many medicinal plants, such as Dendrobium. With the increasing demand for medicinal Dendrobium, the identification of characteristic flavones that can serve as chemical markers for quality control is critical step for quality assurance and safety in the TCM industry. High-performance liquid chromatography coupled with electrospray ionization multi-stage tandem mass spectrometry (HPLC/ESI-MSn ) was used to identify the chemical constituents in five types of Dendrobium: D. crystallinum, D. falconeri, D. strongylanthum, D. moniliforme, and D. gratiosissimum. A total of seventy-six C-glycosyl flavones and three O-glycosyl flavones were identified, of which fifteen C-glycosyl flavones were found in D. crystallinum, twenty four were found in D. falconeri, thirty were found in D. strongylanthum, seven were found in D. moniliforme (also called "Huangtongpi", from Anhui, China), fifteen were found in D. moniliforme (also called "Zitongpi", from Yunnan, China) and seventeen were found in D. gratiosissimum. Additionally, three flavone O-glycosides were all found in D. strongylanthum. The results of this study may be useful for the quality assessment and for the application of D. crystallinum, D. falconeri, D. strongylanthum, D. moniliforme, and D. gratiosissimum. This study provides comprehensive information for identification of flavones from other Chinese herbs.

Quality evaluation of commercial products of Ganoderma lucidum made from its fruiting body and spore

Abstract Ganoderma lucidum (GL), also known as Reishi or Lingzhi, is a medicinal mushroom widely used in traditional and folk medicines. The extracts made from the fruiting body and spore of naturally grown GL are the most frequently used in commercial products. More than 400 compounds have been identified in GL with the triterpenoids considered to be the major active components. Large variations in the chemical components were reported in previous studies and there is no comprehensive study of the content of multiple major triterpenoids in the GL product. In addition, there is no report in the comparison of chemical profiles in different parts of GL (i.e., fruiting body and spore). Determining the chemical composition and comparing the differences between fruiting body and spore are essential for the identity, efficacy and safety of various GL products. In this study, 13 compounds (ganoderenic Acid C, ganoderic Acid C2, ganoderic Acid G, ganoderic Acid B, ganoderenic Acid B, ganoderic Acid A, ganoderic Acid H, ganoderenic Acid D, ganoderic Acid D, ganoderic Acid F, ganoderic Acid DM, ganoderol A, and ergosterol) were selected as the chemical markers. The purpose of this study is to develop an HPLC-DAD fingerprint method for quantification of these active components in GL (spore and fruiting body) and test the feasibility of using the HPLC-DAD fingerprint for quality control or identity determination of GL products. The results showed that this method could determine the levels of the major components accurately and precisely. Among the 13 components, 11 ganoderma acids were identified to be proper chemical markers for quality control of GL products, while ganoderal A was in a very low amount and ergosterol was not a specific marker in GL. The extracts of fruiting body contained more chemical compounds than those of spore, indicating that these 11 compounds could be a better chemical marker for the fruiting body than the spore. The HPLC chemical fingerprint analysis showed higher variability in the quality of GL harvest in different years, while lesser variation in batches harvested in the same year. In conclusion, an HPLC assay detecting 11 major active components and a fingerprinting method was successfully established and validated to be feasible for quality control of most commercial GL products.

Quantification and discovery of quality control chemical markers for Ba-Bao-Dan by UPLC–MS/MS combined with chemometrics

Ba-Bao-Dan (BBD) is a well-known Traditional Chinese medicine (TCM) prescription in China. It was first formulated in approximately 1555 AD. As one of the National Protected TCM, it is widely used to treat jaundice, viral hepatitis, cholecystitis, acute urinary tract infection, cancer, and other diseases. It is a healthcare medicine that is used to prevent many diseases in China. In other Asian countries and in European and American countries, BBD is used as a drug to protect the liver. However, a systematic quality study on BBD chemical markers has not been carried out. This study aimed to establish an ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-MS/MS) method for the quantitative determination of 43 compounds in BBD. Furthermore, the method was used to further find chemical markers for quality control through the combination with chemometrics. The modified chromatographic conditions were achieved on Waters Cortecs C18 column (2.1 × 100 mm, 1.6 μm) with a gradient elution consisting of 0.1 % formic acid in water and acetonitrile with methanol (1:1, V/V). All analytes were determined in the multiple reaction monitoring mode. The method was validated for linearity, detection limits, precision, repeatability, stability and accuracy. The method was used to analyze the 43 compounds in 11 batches of BBD samples. Hierarchical cluster analysis and principal component analysis were applied to evaluate intrinsic quality of BBD and to identify the potential chemical markers for quality control. In conclusion, the method rapidly and sensitively determined the 43 compounds, among which 10 compounds, namely, N-Gin R1, Gin Re, Gin Rg1, Gin Rb1, GCA, Gin Rd, CA, TCA, CDCA, and DCA, were considered as the potential chemical markers for BBD quality control.

Deep eutectic solvents improve the stability of forsythoside A

AbstractForsythoside A is used as the main chemical marker for quality control of Forsythia suspensa (Thunb.) Vahl (F. suspensa, Lianqiao in Chinese) fruits and leaves. The hydrolysis of forsythoside A at high temperature and effects of deep eutectic solvents (DESs) on stability of forsythoside A were studied in the article. Forsythoside A, I, and H may be directly converted into each other at high temperature. With more heating time, they are gradually decomposed into smaller molecular compounds. High temperature, alkaline conditions, high humidity, the oxidant, and light drastically destroyed stability of forsythoside A, while acidic conditions and the antioxidant protected it. Compared with water and alcohol solvents, DESs improved effectively stability of forsythoside A due to the formation of hydrogen bonds between DESs and forsythoside A. Our research preliminarily uncovered the converting mechanism of forsythoside A and confirmed that DES improved the stability of forsythoside A.

Evaluation of VEGF mediated pro-angiogenic and hemostatic effects and chemical marker investigation for Typhae Pollen and its processed product

Ethnopharmacological relevanceTyphae Pollen (TP) is a well-known Traditional Chinese Medicine (TCM) to remove blood stasis. Carbonized Typhae Pollen (CTP), a processed product of TP after being stir-fried, has been widely applied to clinical practice with its capability of hemostasis. However, the underlying mechanism of TP and CTP are still not fully elucidated and discrimination against TP and CTP remains a challenge. Aim of studyThe aim of this study is to investigate whether TP could remove blood stasis by promoting angiogenesis and the process of carbonizing it could enhance hemostatic effect. Meanwhile, some chemical markers for quality control of CTP had better to be found. Material and methodsThe changes of constituents between TP and CTP were analyzed by UPLC-QTOF-MS/MS. We investigated pro-angiogenic and hemostatic effects of TP and CTP in two zebrafish models: VRI-induced ISV insufficiency model and Ator-induced cerebral hemorrhage model. Subsequently, quantitative real-time PCR (qRT-PCR) was applied to investigate the mechanism of pharmacological effects. Finally, chemometric method was applied to find chemical markers. ResultsA total of 19 compounds were identified in qualitative analysis. The loss rate of each compound was calculated and compared. Two compounds (huaicarbon A/B) could only be detected in CTP and the content of flavonoid glycosides in CTP was significantly decreased compared with TP. The average content of the three identified flavonoid aglycones (quercetin, isorhamnetin and kaempferol) was increased about 30 percent in CTP. TP promoted pro-angiogenesis by up-regulating the expression of VEGFA, flt1 and kdr. After heating process, the pro-angiogenic activity was reduced and hemostatic activity was enhanced in CTP. Then qRT-PCR analysis found that CTP could significantly up-regulate the expression of VEGFA and vWF. In the discovery of markers, 6 chemical markers for discrimination of TP and CTP were obtained by chemometric method. ConclusionOur research indicated that the pro-angiogenic activity of TP was involved in VEGF signaling pathway. After processing, hemostatic activity of CTP has been enhanced by up-regulating the expression of VEGFA and vWF. A chemical marker database was established to provide a scientific evidence for quality control, mechanism and the clinical application of TP and CTP.

Discovery of chemical markers for improving the quality and safety control of Sinomenium acutum stem by the simultaneous determination of multiple alkaloids using UHPLC-QQQ-MS/MS

Sinomenium acutum stem is a popular traditional Chinese medicine used to treat bone and joint diseases. Sinomenine is considered the only chemical marker for the quality control of S. acutum stem in mainstream pharmacopeias. However, higenamine in S. acutum stem is a novel stimulant that was banned by the World Anti-Doping Agency in 2017. Therefore, enhancing the quality and safety control of S. acutum stem to avoid potential safety risks is of utmost importance. In this study, a fast, sensitive, precise, and accurate method for the simultaneous determination of 11 alkaloids in S. acutum stem by ultrahigh-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-QQQ-MS/MS) was established. This method successfully analyzed thirty-five batches of S. acutum stem samples. The average contents of sinomenine, magnoflorine, coclaurine, acutumine, higenamine, sinoacutine, palmatine, magnocurarine, columbamine, 8-oxypalmatine, and jatrorrhizine were 24.9 mg/g, 6.35 mg/g, 435 μg/g, 435 μg/g, 288 μg/g, 44.4 μg/g, 22.5 μg/g, 21.1 μg/g, 15.8 μg/g, 9.30 μg/g, and 8.75 μg/g, respectively. Multivariate analysis, including principal component analysis (PCA), orthogonal partial least square method-discriminant analysis (OPLS-DA), and hierarchical cluster analysis (HCA), were performed to characterize the importance and differences among these alkaloids in S. acutum stem samples. As a result, sinomenine, magnoflorine, coclaurine, acutumine, and higenamine are proposed as chemical markers for quality control. Higenamine and coclaurine are also recommended as chemical markers for safety control. This report provides five alkaloids that can be used as chemical markers for improving the quality and safety control of S. acutum stem. It also alerts athletes to avoid the risks associated with consuming S. acutum stem.

Identification and quantification of oleanane triterpenoid saponins and potential analgesic and anti-inflammatory activities from the roots and rhizomes of Panax stipuleanatus

BackgroundPanax stipuleanatus represents a folk medicine for treatment of inflammation. However, lack of experimental data does not confirm its function. This article aims to investigate the analgesic and anti-inflammatory activities of triterpenoid saponins isolated from P. stipuleanatus. MethodsThe chemical characterization of P. stipuleanatus allowed the identification and quantitation of two major compounds. Analgesic effects of triterpenoid saponins were evaluated in two models of thermal- and chemical-stimulated acute pain. Anti-inflammatory effects of triterpenoid saponins were also evaluated using four models of acetic acid–induced vascular permeability, xylene-induced ear edema, carrageenan-induced paw edema, and cotton pellet–induced granuloma in mice. ResultsTwo triterpenoid saponins of stipuleanosides R1 (SP-R1) and R2 (SP-R2) were isolated and identified from P. stipuleanatus. The results showed that SP-R1 and SP-R2 significantly increased the latency time to thermal pain in the hot plate test and reduced the writhing response in the acetic acid–induced writhing test. SP-R1 and SP-R2 caused a significant decrease in vascular permeability, ear edema, paw edema, and granuloma formation in inflammatory models. Further studies showed that the levels of inflammatory mediators, nitric oxide, malondialdehyde, tumor necrosis factor-α, and interleukin 6 in paw tissues were downregulated by SP-R1 and SP-R2. In addition, the rational harvest of three- to five-year-old P. stipuleanatus was preferable to obtain a higher level of triterpenoid saponins. SP-R2 showed the highest content in P. stipuleanatus, which had potential as a chemical marker for quality control of P. stipuleanatus. ConclusionThis study provides important basic information about utilization of P. stipuleanatus resources for production of active triterpenoid saponins.

Simultaneous Quantitative Determination of Polyphenolic Compounds in Blumea balsamifera (Ai-Na-Xiang, Sembung) by High-Performance Liquid Chromatography with Photodiode Array Detector.

A high-performance liquid chromatography method was developed for simultaneous quantification of 18 polyphenolic compounds from the leaves of Blumea balsamifera, including 17 flavonoids and 1 phenylethanone. The B. balsamifera extraction was separated by a Kromasil C18 column (250 × 4.6 mm, 5 μm) with a binary gradient mobile phase consisting of acetonitrile and 0.2% aqueous acetic acid. A photodiode array detector (PDA) was used to record the signals of investigated constituents. The linearity, sensitivity, stability, precision, and accuracy of the established assay methods were assessed to meet the requirements of quantitative determination. Samples extracted by reflux in 25 mL of 80% methanol for 30 minutes were selected for the extraction method. The 18 compounds were accurately identified by comparing with the reference compounds. The purity of each peak was confirmed by the base peak in the mass spectrum. The contents of 18 compounds in Blumea samples from four different regions were successfully determined. The results also showed that 3,3′,5,7-tetrahydroxy-4′-methoxyflavanone was the most abundant constituent, which could be used as a potential chemical marker for quality control of B. balsamifera and Chinese patent medications containing B. balsamifera herb.

Discovery of chemical markers for identifying species, growth mode and production area of Astragali Radix by using ultra-high-performance liquid chromatography coupled to triple quadrupole mass spectrometry

BackgroundAstragali Radix (AR) is a well-known Chinese herbal medicine. The quality of AR can be affected by many factors such as species, growth mode and production area, but there are still no chemical markers to distinguish it. PurposeTo explore chemical markers for improving the quality assessment of AR and discover chemical markers for identifying species, growth mode and production area of AR. MethodsA highly sensitive, efficient and accurate method based on ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) for simultaneous quantitative determination of 14 major chemical components (five flavonoids and nine triterpene saponins) in 94 batches of AR from China, Republic of Korea and Germany was developed for the first time. To explore chemical markers and assess changes in the contents of 14 compounds in the 94 batches of AR samples from different regions, hierarchical clustering analysis (HCA) and principal component analysis (PCA) were performed. ResultsAstragaloside III was not only an important chemical marker for distinguishing two species of AR, i.e.: Astragalus mongholicus and A. membranaceus, but also a potential chemical marker for the classification of cultivated and semi-wild AR. In addition, in the batches of cultivated AR, the content of isoastragaloside II and cyclocephaloside II were greater in batches from the region of Shaanxi Province than that of other Provinces in China, but the content of calycosin-7-O-β-D-glucoside and astragaloside IV, which are the quality control markers of AR required by the Chinese Pharmacopoeia, were higher than that of other Provinces in China. In addition, the content of calycosin-7-O-β-D-glucoside, ononin, calycosin and astragaloside I could be used to identify samples of AR collected from China, Republic of Korea and Germany. ConclusionThis UHPLC-QQQ-MS/MS method could be applied to the quantitative evaluation of AR and could be an important and meaningful reference to develop chemical markers for quality control of AR.

Investigation on apposite chemical marker for quality control of Tephrosia purpurea (L.) Pers. by means of HPTLC-chemometric analysis

Application of phytochemicals as the markers for quality assurance of the herbal medicinal material is one of the prominently used approach. In the present study, six major chemical compounds i.e. rutin, quercetin, lupeol, β-sitosterol, rotenone, deguelin of therapeutically important plant Tephrosia purpurea were tested for their significance to be used as chemical markers in analytical methods. Plants were collected from different locations. Extraction procedures as well as HPTLC analytical methods have been optimized for each compound. All these methods have been validated in terms of linearity, intraday-interday precision, LOD, LOQ, accuracy and repeatability. Metabolites have been quantified and quantitative data has been subjected to chemometric analysis. Results revealed that Rf values of all the compounds are between 0.3 and 0.4. Rutin, lupeol and β-sitosterol gave desirable response and other three compounds were found undetectable in HPTLC. Content of rutin, beta-sitosterol and lupeol ranged from 0.095 to 0.84, 0.043 to 0.125, 0.023 to 0.045 w/w % respectively. Among them, rutin content was highest in all samples. Quantitative measurements combined with chemometric analysis displayed chemodiversity in the samples. In addition, principal component analysis ranked the marker as in order of their significance rutin > β-sitosterol > lupeol. Results indicate rutin to be most preferable chemical marker for the Tephrosia. Furthermore, application of all the three compounds combined as the multimarker system should be preferred for standardization of T. purpurea.

EFFECT OF ETHANOL ON THE DRYING CHARACTERISTICS AND ON THE COUMARIN YIELD OF DRIED GUACO LEAVES (Mikania laevigata SCHULTZ BIP. EX BAKER)

Abstract Guaco is a common medicinal plant in Brazil used for the treatment of respiratory diseases. Coumarin serves as a chemical marker for quality control of products derived from guaco. In this study, the drying kinetics and coumarin yield of guaco leaves dried with and without treating the surface with ethanol were evaluated. The drying experiments were carried out in a forced convection tunnel at 50 and 60 oC temperatures and air velocities of 0.42 and 0.84 m/s. The results show that the samples with ethanol experienced faster water evaporation during drying experiments, thereby reducing the drying time by up to 35%, and also had higher yields of coumarin. Storage experiments and color analysis were also performed. Treatment of dried guaco leaves with ethanol resulted in color degradation, but increased shelf life by 20%.

Chemical markers for quality control of bran-fried sulfur-fumigated Paeoniae Radix Alba

In traditional Chinese medicine practice, crude herbs are often subjected to traditional processing (Paozhi in Chinese) for a special medicinal purpose. Bran-frying is one of processing methods for Paeoniae Radix Alba (PRA). Previous studies found that paeoniflorin and paeoniflorin sulfonate, a principle bioactive compound and its sulfur-fumigation induced characteristic sulfur-containing derivative, could be used together with sulfur dioxide residue as chemical markers for the quality control of sulfur-fumigated PRA crude material. In this paper, the feasibility of these three markers used for the quality control of bran-fried sulfur-fumigated PRA was further investigated. First, homemade samples of sulfur-fumigated PRA with different sulfur-fumigation duration (0.5–6 h) were bran-fried, and stored for 12 months. Second, the contents of sulfur dioxide residue, paeoniflorin and paeoniflorin sulfonate were dynamically quantified respectively. Third, the variation of the marker contents and their correlation during bran-frying and storage was evaluated. A validation was conducted using commercial bran-fried PRA samples. The results showed that bran-frying caused an averaged reduction of 20% in the content of sulfur dioxide residue, and during the first two months of storage the content of sulfur dioxide residue was decreased by up to 27%, then the content was tending towards stability for the subsequent ten months of storage (RSD = 3.92%). Meanwhile, paeoniflorin and paeoniflorin sulfonate were relatively stable, the contents of which were not affected by bran-frying processing and 12 months of storage. The correlations between the contents of sulfur dioxide residue and paeoniflorin/paeoniflorin sulfonate were obviously influenced by storage duration. Since sulfur dioxide residue is a safety marker, while paeoniflorin and paeoniflorin sulfonate can reflect the inner quality and the impact extent of sulfur-fumigation on the quality of bran-fried PRA respectively, these three chemicals might be used together as markers for the quality control, and consequently to ensure the safety and efficacy of bran-fried PRA.

Molecular Markers Associated With Chemical Analysis: A Powerful Tool for Quality Control Assessment of Copalchi Medicinal Plant Complex.

The copalchi complex, Hintonia latiflora, H. standleyana, and Exostema caribaeum, is widely used in Mexico for treating diabetes and gastrointestinal disorders. The first therapeutic use for H. latiflora bark was registered in the “Florentine Codex” in the sixteenth century. The latest pharmacological and phytochemical studies revealed that the infusion of the leaves have hypoglycemic, antihyperglycemic and gastroprotective activities. For these reasons the monograph of the main copalchi species, H. latiflora, was recently added to the Mexican Herbal Pharmacopoeia. Nevertheless, quality control parameters are focused to the bark but not to the leaves. Moreover, information about other Rubiaceae species is needed. The main goal of this study was to generate molecular and chemical markers for quality control of the copalchi complex raw material. In addition, the resolution of the taxonomical ambiguity between H. latiflora and H. standleyana, as well as the testing of the molecular and chemical markers in different geographical batches, were aims of this study. The molecular markers and chemical profiles of the leaf infusions were generated considering three different populations for H. latiflora and separate individuals of the three species (HL, n = 10; HS, n = 3; EC, n = 4). The molecular markers matK, rbcL, trnH-psbA, rpl32-trnL, and ITS2 were tested for their discriminating capabilities. Chemical profiles of the leaf infusions were obtained by means of HPLC analyses using chlorogenic acid and 4-phenylcoumarins as chemical markers. The concatenated sequence of the molecular markers trnH-psbA, rpl32-trnL, and ITS2 clearly distinguished the three taxa, clarifying the taxonomical ambiguity of the Hintonia genus. Additionally, the chemical profiles allowed the unequivocal identification of each species supporting the molecular results; the geographical origin of the samples did not modify neither the chemical profiles nor the concatenated sequence of H. latiflora, suggesting that it is a robust identity test. The complementary use of molecular and chemical markers will assure the quality of plant material used in traditional medicine for therapeutic purposes, and should be valuable new information for the National Health authorities as a part of the Mexican Herbal Pharmacopoeia.