Chemical Profile Characterization Research Articles

Characterization of chemical profiles in Cannabis sativa: evaluation of a popular extraction method to identify chemotypes

Este estudio aborda la caracterización de los cannabinoides de distintas variedades de Cannabis sativa cultivadas en Paraguay, enfocándose en la determinación de la relación entre los fitocannabinoides primarios, tetrahidrocannabinol (THC) y cannabidiol (CBD). Utilizando técnicas avanzadas de cromatografía líquida de alta resolución (HPLC), se analizaron los extractos de Cannabis, identificando variaciones significativas en las concentraciones de cannabinoides que subrayan la diversidad química intrínseca entre las variedades estudiadas. Además, se evaluaron métodos de extracción caseros, empleando etanol y maceración en frío, seguidos de una evaporación del solvente a baja temperatura. Los resultados mostraron relaciones variables de CBD: THC, indicando la idoneidad de los extractos para aplicaciones terapéuticas e industriales (cáñamo o Cannabis no psicoactivo). La investigación se llevó a cabo teniendo en cuenta las regulaciones nacionales, garantizando la integridad y aplicabilidad de los hallazgos. Este trabajo no solo contribuye al corpus científico existente sobre el Cannabis, sino que también resalta la importancia de la investigación orientada a la optimización de su cultivo y utilización medicinal, alineada con los intereses nacionales expresados en el Decreto N° 3999.

Chemical Profile Characterization of Fruit and Vegetable Juices after Fermentation with Probiotic Strains.

Lactic acid bacteria (LAB) are widely applied for fermentation purposes in dairy and non-dairy food matrices with beneficial technological and health-promoting properties. This study describes the effect of two lactic acid bacteria, namely, Lactiplantibacillus paracasei SP5 and Pediococcus pentosaceus SP2, on the phenolic profiles, antioxidant activities, total phenolic content (TPC), carotenoid content, and sensorial profile of two different mixed fruit juices. After 48 h of fermentation, both LABs retained viability over 9 Log CFU/mL in both juices. The TPC, zeaxanthin + lutein, β-carotene content, and antioxidant activity (AA) were elevated for both LABs and mixed juices after 48 h of fermentation compared to control samples. Regarding the phenolic profile, both juices exhibited a significant decrease in chlorogenic acid levels, while quinic acid and tyrosol concentrations showed notable increases.

Integrated post-acquisition data processing strategy for rapid steroid sulfate characterization in Toad gall-bladder

LC–MS serves as a workhorse for chemical profile characterization of Chinese medicinal materials (CMMs) attributing to the ability of measuring fruitful MS/MS spectral information. However, it is laborious to extract the information belonging to the compounds-of-interest from the massive data matrixes even employing those well-defined post-acquisition data processing strategies. Here, efforts were devoted to propose an integrated strategy allowing rapid chemical homologs-focused data filtering through integrating the fit-for-purpose existing strategies, such as molecular weight imprinting (MWI), diagnostic fragment ion filtering (DFIF), neutral loss filtering (NLF), and isotope pattern filtering (IPF). Homologs-focused chemical characterization of a precious CMM namely Toad gall-bladder (Chinese name: Chandan) that is rich of diverse effective steroid sulfates, particularly bufogenin sulfates, bile acid sulfates and bilichol sulfates, was employed as a proof-of-concept. Recombinant human SULT2A1-catalyzed in vitro metabolism was undertaken to generate eight bufogenin sulfates to facilitate summarizing MS/MS spectral behaviors. After in-house data library construction and MS1 and MS2 spectral acquisition, data filtering was conducted as follows: 1) MWI and IPF was utilized in combination to capture deprotonated molecular ions and the 34S isotopic ions for the sulfates of those reported steroids; 2) m/z 79.9568 (SO3−·) and 96.9596 (HSO4−) were applied to DFIF; and 3) SO3 (79.9568 Da) served as the feature to achieve NLF. Those captured MS/MS information subsequently participated in tentatively structural annotation through applying those empirical mass fragmentation rules. As a result, 71 compounds including 7 bufogenin sulfates, 17 bile acid sulfates, 13 bilichol sulfates and a C-23 steroid sulfate were detected from Toad gall-bladder and thereof, 39 ones received plausible identities assignment. Above all, the steroid sulfates in Toad gall-bladder were profiled in depth, and more importantly, the proposed strategy should be a meaningful option for, but not limited to, submetabolome characterization in CMMs.

GC/MS and 2D NMR-based approach to evaluate the chemical profile of hydroalcoholic extract from Agaricus blazei Murill and its anti-inflammatory effect on human neutrophils

Ethnopharmacological relevanceAgaricus blazei Murill (AbM) is one of the main mushrooms used for medicinal purposes. The use of AbM in the preparation of teas is widespread mainly in Asian countries, while in Brazil it is used as a functional food to combat inflammatory diseases and cancer. Aim of the studyThe main focus of this study was the characterization of the chemical profile of the hydroalcoholic extract of Agaricus blazei Murill (AbE), as well as the evaluation of its cytotoxic and anti-inflammatory potential using human neutrophils. Materials and methodsThe extract was prepared by dynamic maceration using a mixture of ethanol and water (70/30, v v−1) as solvent. The chemical profile characterization was carried out by 2D NMR and GC-MS techniques. The cytotoxicity of AbE was evaluated through studies of hemolytic potential, cell viability and membrane integrity. The anti-inflammatory activity was analyzed by a PMA-induced neutrophil degranulation assay. ResultsChemical analysis of AbE revealed the presence of 28 metabolites in its composition, with mannitol as the major compound. AbE at 1–200 μg mL−1 and mannitol at 4–160 μg mL−1, showed low hemolytic and cytotoxic potential against human red blood cells and neutrophils. Furthermore, both were able to significantly reduce the release of myeloperoxidase. ConclusionsThese results indicate that AbE is a promising natural product to be incorporated into pharmaceutical dosage forms intended for the adjuvant treatment of inflammatory diseases.

Chemical profiling and mechanistic studies of Zhi-Shang-Feng granules against influenza virus by high-performance liquid chromatography coupled with Q exactive focus hybrid quadrupole orbitrap high-resolution mass spectrometry in combination with network pharmacology analysis.

Zhi-Shang-Feng Granules are used in the clinical treatment of influenza to relieve headaches, chills and fever, bronchitis, nasal congestion, neuralgia and other symptoms. To decipher the components responsible for therapeutic effects of Zhi-Shang-Feng g ranules against influenza virus, an analytical method based on high-performance liquid chromatography coupled with Q exactive focus hybrid quadrupole orbitrap high resolution mass spectrometry was developed and the chemical profile of Zhi-Shang-Feng granules was characterized. Then, the identified components were used to conduct network pharmacological analysis and determine the potential mechanism of Zhi-Shang-Feng Granules. As a result, 177 compounds were putatively identified through comprehensive analysis by liquid chromatography coupled with high-resolution mass spectrometry, of which 23 compounds were unambiguously confirmed with reference standards. Components in Zhi-Shang-Feng Granules were found to specifically act on different enzymes, G-protein-coupled receptors, ion channels and transporters in the immune, endocrine, nervous, and circulatory systems. The potential mechanism was related to several biological processes, including cell growth and death, pattern recognition receptor signalling, signalling by interleukins, and lipid metabolism. The combination of chemical profile characterization and network construction provided useful insight into the overall chemical composition of Zhi-Shang-Feng granules and revealed their potential anti-infection, anti-inflammatory and immunoregulatory mechanisms against influenza virus infected disease.

Optimization of Ultrasonic-Assisted Total Flavonoids Extraction from Flower of Albizia Julibrissin Durazz. by Response Surface Methodology for Characterization of Chemical Profile and Evaluation of Antioxidant and Tyrosinase Inhibitory Activities

Optimization of Ultrasonic-Assisted Total Flavonoids Extraction from Flower of <i>Albizia Julibrissin</i> Durazz. by Response Surface Methodology for Characterization of Chemical Profile and Evaluation of Antioxidant and Tyrosinase Inhibitory Activities

Physicochemical Characterization and Paper Spray Mass Spectrometry Analysis of Myrciaria Floribunda (H. West ex Willd.) O. Berg Accessions.

Myrciaria floribunda, also known as rumberry, is a tree native to the Brazilian Atlantic Forest, where its fruits have the potential for commercial use. This study evaluates the antioxidant potential, determines the phytochemical profile, and chemically characterizes the rumberry fruit. Accessions were sampled from the Rumberry Active Germplasm Bank of the Federal University of Alagoas, Brazil. Physical characteristics, chemical characteristics, and phenolic compound content were analyzed. Chemical profile characterization was carried out using PSMS. Accessions had an average weight of 0.86 g. Accession AC137 presented a higher pulp yield (1.12 g). AC132 and AC156 had larger fruits, AC137 showed greater firmness (5.93 N), and AC160 had a higher and total phenolic content ratio (279.01 ± 11.11). Orange-colored accessions scored higher in evaluated parameters, except for AC160 for phenolic content. Thirty-two compounds were identified on positive ionization mode and 42 compounds on negative ionization mode using PSMS. Flavonoids, followed by the derivatives of benzoic acid, sugars, and phenylpropanoids, were the most prominent. Myricitrin, quercitrin, and catechin stand out as flavonoids that have been reported in previous studies with antioxidant and antimicrobial properties, in addition to health and therapeutical benefits, demonstrating the potential of the rumberry fruit.

Rapid chemome profiling of Artemisia capillaris Thunb. using direct infusion-mass spectrometry

As one of the most popular traditional Chinese medicines (TCMs) for the treatment of various liver diseases, virgate wormwood herb ( Artemisia capillaris Thunb.) has a long application history in TCM practices. It has been well established that the chemical composition is responsible for the pronounced therapeutic spectrum of A. capillaris . Although they are comprehensive, the time-intensive liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) assays cannot fully satisfy the analytical measurement workload from many test samples. Direct infusion-MS/MS (DI–MS/MS) may be the optimal choice to achieve high-throughput analysis if the mass spectrometer can universally record MS 2 spectra. According to the application of gas phase ion fractionation concept, the MS/MS ALL program enables to gain MS 2 spectrum for each nominal m/z value with a data-independent acquisition algorithm via segmenting the entire MS 1 ion cohort into sequential ion pieces with 1 Da width, when sufficient measurement time is allowed by DI approach. Here, rapid clarification of the chemical composition was attempted for A. capillaris using DI–MS/MS ALL . A. capillaris extract was imported directly into the electrospray ionization interface to obtain the MS/MS ALL measurement. After the MS 1 -MS 2 dataset was well organized, we focused on structural characterization through retrieving information from the available databases and literature. Twenty-six compounds were found, including 12 caffeoyl quinic acid derivatives, 7 flavonoids, and 7 compounds belonging to other chemical families. Among them, 24 ones were structurally identified. Compared with the LC–MS/MS technique, DI–MS/MS ALL has the advantages of low-costing, solvent-saving, and time-saving. Chemical profiling of A. capillaris extract was accomplished within 5 min by DI–MS/MS ALL , and this technique can be an alternative choice for chemical profile characterization of TCMs due to its extraordinary high-throughput advantage.

Analysis of chemical components of Chinese medicine Ligustici Radix by achiral-chiral liquid chromatography-predictive multiple reaction monitoring

中药毛前胡为伞形科植物短片藁本Ligusticum brachylobum Franch.的干燥根,主要用于治疗风热咳嗽痰多、痰热喘满、咯痰黄稠等证,富含香豆素类化学成分,含有多组对映异构体和非对映异构体。为了深入研究毛前胡的化学成分组成,特别是对映异构体的组成,研究建立了非手性-手性色谱-预测多反应监测法(achiral-chiral-LC predictive MRM),同步实现毛前胡化学成分的化学选择性和立体选择性分离,以及高灵敏度定性分析。非手性色谱和手性液相色谱-串联质谱系统结合了RP-C18色谱柱的高效化学选择性分离能力以及手性色谱柱的立体选择性优势,有效避免了中心切割非手性-手性二维液相色谱构造复杂、重现性难以满足定量要求等缺陷。采用小内径核-壳型RP-C18色谱柱作为前端化学分离柱,实现结构类似香豆素的高效化学选择性分离;采用反相大内径AD-RH手性色谱柱,实现对映异构体的手性拆分;采用预测多反应监测模式,实现化学成分的高灵敏度检出;利用增强子离子扫描模式(EPI)采集各色谱峰的二级质谱信息,鉴定化学结构。通过定量离子对、定性离子对及两者的比值,判定是否为对映异构体。利用所构建的非手性-手性色谱耦联系统从毛前胡中共鉴定出60个化学成分,其中8对香豆素对映异构体得到了良好分离。本研究为毛前胡以及含有对映异构体中药的深入定性、定量分析提供可靠的方法。

Chemical profile and potential mechanisms of Huo-Tan-Chu-Shi decoction in the treatment of coronary heart disease by UHPLC-Q/TOF-MS in combination with network pharmacology analysis and experimental verification

Huo-Tan-Chu-Shi Decoction (HTCSD), a traditional Chinese medicine (TCM) prescription within Guangdong Provincial TCM Hospital (the largest TCM hospital in China), is used for effective clinical treatment of coronary heart disease (CHD) caused by phlegm-dampness syndrome with high incidence in the hot and humid climate of Lingnan region. However, its chemical components responsible for the therapeutic effects remain unclear, which restricts its application and further development. Hence, a detailed workflow, combing with UHPLC-Q/TOF-MS, network pharmacology analysis and experimental verification, was proposed and applied to characterize the chemical profile and potential mechanism of HTCSD against CHD. As a result, a total of 130 components from all six composed herbal medicines were characterized in a rapid and sensitive manner through UHPLC-Q/TOF-MS, of which 33 compounds were unambiguously confirmed with reference standards. Consequently, based on the integrated pharmacology network of “herbs-chemicals-targets-pathways-therapeutic effects”, four chemicals (magnoflorine, menisperine, 13-hydroxyberberine, luteolin) with four CHD related targets (SRC, MAPK1, EGFR and AKT1) were considered as the key components and targets of HTCSD in the treatment of CHD. Furthermore, the effect of HTCSD was confirmed in animal experiments by enhancing the phosphorylation of MAPK, and the published literature and molecular binding results suggested that magnoflorine and luteolin tended to be the critical compounds involved in the process. Taken together, the characterization of chemical profile combined with network pharmacology analysis and experimental verification not only provided an efficient insight into the overall chemical profile of HTCSD but also revealed the potential pharmacological components and mechanisms of HTCSD against CHD, which laid a necessary chemical and biological basis for the discovery of in vivo bioactive components and the further revelation of functionary mechanism.

Characterization of chemical profile and quantification of major representative components of Wendan decoction, a classical traditional Chinese medicine formula.

Wendan decoction, a classical traditional Chinese medicine formula consisting of six herbal medicines, has been widely used in clinical treatments for thousands of years due to the expectorant effects. However, the chemical basis of Wendan decoction remains unclear, which hinders the elucidation of the scientific connotation and mechanism of its effective components. In this study, an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry method was first developed for characterization of its chemical profile, and a total of 142 chemical components including flavonoids, triterpenoids, alkaloids, coumarins, pungent phytochemicals, and other types were detected, among which 41 components were definitively identified with authentic standards. Furthermore, 14 major representative components were simultaneously quantified by high-performance liquid chromatography with ultraviolet detector, indicating that the content levels of flavonoids were the most abundant in Wendan decoction. In summary, this study established sensitive and practical methods to systematically characterize chemical profile for the first time and simultaneous quantify representative components of Wendan decoction. These findings above would provide a solid chemical basis for disclosure of potential effective components by further in vivo disposal study, and promote therapeutic mechanism researches of Wendan decoction.

Hatching Induction of Cyst Nematodes in Bare Soils Drenched With Root Exudates Under Controlled Conditions.

Cyst nematodes account for substantial annual yield losses in crop production worldwide. Concerns over environmental and health issues due to the use of chemical nematicides mean alternative sustainable and integrated solutions are urgently required. Hatch induction of encysted eggs in the absence of host plants, i.e., ‘suicide-hatching,’ could be a sustainable alternative in reducing population densities of cyst nematodes in infested soils. Here we examined in situ hatching of encysted eggs of Globodera pallida, Heterodera carotae, and Heterodera schachtii at varying soil depths, following exogenous applications of host root exudates in repeated glasshouse experiments. Cysts were retrieved 30 or 43 days post-incubation depending on the nematode species and assessed for hatching rates relative to the initial number of viable eggs per cyst. Hatching of the potato cyst nematode G. pallida depended on both soil moisture and effective exposure to root exudates, and to a lesser extent on exudate concentration. The carrot cyst nematode H. carotae had over 75% hatched induced by root exudate irrespective of the concentration, with better hatch induction at 20 cm as compared with 10 cm soil depth. Hatching of the beet cyst nematode H. schachtii largely depended on the soil moisture level at constant temperature, rather than the type or concentration of root exudates applied. As a conclusion, exogenously applied host root exudates may play a major role in inducing in situ hatch of encysted eggs of potato and carrot cyst nematodes in the absence of host plant under favorable soil temperature/moisture conditions. To improve such strategy, the characterization of chemical profiles of the root exudate composition and field validation are currently ongoing.

Bioactive activities and chemical profile characterization using paper spray mass spectrometry of extracts of Eriobotrya japonica Lindl. leaves.

The loquat (Eriobotrya japonica Lindl.) is a fruit tree that has been used in Chinese medicine for thousands of years for the treatment of various diseases. The loquat leaf extracts contain several bioactive compounds with antioxidant and antimicrobial properties, and identification of these substances using quick and simple methods has been an analytical trend. The influence of dehydration of loquat leaves (without drying, at 40°C, and at 60°C), the type of solvent (ethanol and methanol), and the method of extraction (shaking and ultrasound) on obtaining extracts containing phenolic compounds and substances with antioxidant and antimicrobial properties was evaluated. The chemical constituents of an extract were identified using paper spray mass spectrometry (PS-MS). The extract obtained from the dehydrated leaves at 40°C presented the best results. The extracts obtained from these leaves and with ethanol had the highest values of total phenolics and antioxidant activities, but the methanolic extract subjected to ultrasound had the highest levels of chlorogenic, caffeic, and ellagic acids. All extracts evaluated inhibited the growth of Staphylococcus aureus. Using the PS-MS technique, it was possible to identify the presence of 49 substances such as organic acids, phenolic acids, flavonoids, sugars, quinones, and terpenes. In general, extracts of dehydrated leaves at 40°C and extracted with ethanol using ultrasound can be considered a good source of bioactive compounds with potential applications as functional ingredients or additives in the food and pharmaceutical industries. PS-MS was demonstrated to be a simple and ultrafast technique to obtain the chemical profile of the loquat leaf extract.

Characterization of chemical profiles of pH-sensitive cleavable D-gluconhydroximo-1, 5-lactam hydrolysates by LC–MS: A potential agent for promoting tumor-targeted drug delivery

Currently, controllable linker cleavage at the target site will facilitate the clinical treatment of cancer. Dual-functional prodrugs in combination of carbohydrate as targeting group and pH-sensitive cleavable linker are desired in clinical development. Here, a qualified structure of N-phenylcarbamate-d-gluconhydroximo-1,5-lactam was employed and proved to be a potential candidate prodrug in the drug design. To proof this concept, the possible mechanism of Beckmann rearrangement and the degraded products were confirmed by HPLC and LC–MS under the acid condition mimic lysosome. Hence, the strategy of d-gluconhydroximo-1,5-lactam as a prodrug carrier fabricated with interested drugs will provide a great potential approach for chemotherapy.

Characterization of chemical profile and quantification of representative components of DanLou tablet, a traditional Chinese medicine prescription, by UHPLC-Q/TOF-MS combined with UHPLC-TQ-MS

DanLou tablet (DLT), a famous traditional Chinese medicine prescription (TCMP) consisting of 10 herbal medicines, is extensively used for the treatment of angina pectoris and acute coronary syndrome in China. However, active chemical constituents responsible for the therapeutic effects still remain unclear, due to the fact that the complex composition in DLT have not been holistically clarified. Therefore, this study aimed to characterize the chemical profile and simultaneously quantify the representative components in DLT. First, 157 chemical constituents including flavonoids, triterpenoids, tanshinones, lactones, phenolic acids, paeoniflorins and the other types of components were detected, among which 39 were exactly identified by comparing their retention times and MS fragmentation behaviors with those of authentic standards by ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q/TOF-MS). Moreover, 33 representative components were simultaneously quantified by ultra-high performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UHPLC-TQ-MS), which were selected based on following three principles: qualitative and quantitative markers in the Chinese Pharmacopeia (2015 edition), bioactive components possessing cardiovascular-related in vivo or in vitro activities and those derived from 10 consisted herbs in DLT with a diversity of representative structure types. The method was validated in terms of linearity, precision, repeatability and recovery and successfully applied for the quality evaluation of 20 batches of DLT samples. Further chemometric analysis indicated that danshensu and salvianolic acid B were the most significant quantitative markers for the content fluctuation of DLT. In summary, the chemical profiles of DLT were systematically characterized and a practical quantitative method combined with chemometrics was developed to evaluate the intrinsic quality of multiple DLT samples in this study. The present work would be helpful for guaranteeing the safety, efficacy, and controllability in clinical medication of DLT.

A Bioactive Chemical Markers Based Strategy for Quality Assessment of Botanical Drugs: Xuesaitong Injection as a Case Study

Current chemical markers based quality assessment methods largely fail to reflect intrinsic chemical complexity and multiple mechanisms of action of botanical drugs (BD). The development of novel quality markers is greatly needed. Here we propose bioactive chemical markers (BCM), defined as a group of chemo-markers that exhibit similar pharmacological activities comparable to the whole BD, which can therefore be used to effectively assess the quality of BD. As a proof-of-concept, a BCM-based strategy was developed and applied to Xuesaitong Injection (XST) for assessing the efficacy and consistency of different batches. Firstly, systemic characterization of chemical profile of XST revealed a total number of 97 compounds. Secondly, notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and Rd were identified as BCM of XST on treating cardiovascular and cerebrovascular diseases according to Adjusted Efficacy Score following an in vivo validation. Analytical method for quantification of BCM was then developed to ensure the efficacy of XST. Finally, chemical fingerprinting was developed and used to evaluate the batch-to-batch consistency. Our present case study on XST demonstrates that BCM-based strategy offers a rational approach for quality assessment of BD and provides a workflow for chemistry, manufacturing, and controls (CMC) study of BD required by regulatory authority.

Chemical Profile Characterization of Artemisia annua L. Essential Oils From South India Through GC-FID and GC-MS Analyses

:This investigation examined the essential oils isolated from the shoot biomass of Artemisia annua var. Jeevanraksha, a medicinal-cum-aromatic plant grown in Bangalore (Karnataka) and Hyderabad (Andhra Pradesh), south India. GC-FID, GC-MS analyses revealed the presence of 51 (Bangalore) and 41 (Hyderabad) compounds accounting for 93.8 % and 93.9 % of the essential oils, respectively. The principal constituents of Bangalore and Hyderabad essential oils, respectively were: myrcene (8.3 %, 15.4 %), 1,8- cineole (9.7 %, 17.2 %) and camphor (42.6 %, 34.2 %). Other components were: α-pinene (2.8 %, 2.6 %), camphene (3.4 %, 3.9 %), sabinene (1.6 %, 2.7 %), artemisia alcohol (0.7 %, 4.3 %), camphene hydrate (0.5 %, 4.1 %), terpinen-4-ol (1.1 %, 0.2 %), α-terpineol (1.0 %, 1.3 %), β-caryophyllene (2.3 %, 0.4 %), β-selinene (3.9 %, 0.4 %) and β-eudesmol (1.7 %, 0.2 %). Bangalore essential oil had higher percentages of aliphatic ketones (43.3 %), sesquiterpene hydrocarbons (9.9 %) and oxygenated sesquiterpenes (4.3 %). Hyderabad essential oil contained greater concentrations of monoterpene hydrocarbons (25.7 %) and oxygenated monoterpenes (65.5 %).

Chemical profile characterization of biologically active extracts obtained from Salvia species

Chemical profile characterization of biologically active extracts obtained from Salvia species