[Chemome profiling of Qijiao Shengbai Capsules by UPLC-IT-TOF-MS].

Qijiao Shengbai Capsules(QJ) are a common Miao medicine serving as an adjuvant cancer therapy in clinical practice.QJ consists of seven medicinal materials such as Astragalus membranaceus and Lespedeza buergeri.Its chemical components have not been clarified and the quality control needs to be improved.In this study, LC-IT-TOF-MS was used to comprehensively collect MS~1 and MS~2 fragment information of QJ and rapidly identify the chemical compositions.The chromatographic separation was performed on the Capcell core ADME column(2.1 mm×150 mm, 2.7 µm) with 0.1% formic acid aqueous solution(A) and acetonitrile(B) as mobile phases for gradient elution.High-resolution mass spectrometric information was obtained by scanning in the positive and negative ion ESI modes.A total of 107 compounds were structurally identified according to the deduced MS fragmentation patterns and comparison with standards and data reported in the literature, including 54 flavonoids, 16 phthalides, 13 alkaloids, 12 phenolic acids, 7 saponins, 2 coumarins, 2 condensed tannins, and 1 purine.This study clarified the chemical composition of QJ and provided references for the improvement of its quality standards and the elucidation of its medicinal substances.

[Rapid chemome characterization of Pien-Tze-Huang by DI-MS/MS~(ALL)].

Pien-Tze-Huang, one of the most famous and widely used Chinese medicinal prescriptions in China, consists of Notoginseng Radix et Rhizoma, Bovis Calculus, Fel Serpentis, and Moschus.The prescription can clear heat and remove toxin, cool blood and resolve blood stasis, and relieve swelling and pain.Characterizing the chemical composition can facilitate the construction of the quality standard and the research on the effective compounds and action mechanism of Pien-Tze-Huang.Therefore, this study used direct infusion(DI)-MS/MS~(ALL) method to rapidly and accurately reveal the chemical composition of Pien-Tze-Huang.The principle of chemical composition profiling of Chinese medicinal prescriptions lies in the MS~1-MS~2 dataset construction, followed by structural annotation based on MS/MS spectra and summarizing of mass fragmentation pathways.MS/MS~(ALL) owns unique mass spectrometric separation ability via applying gas phase fractionation which enables MS~1 ion cohort successively enter the collision cell and acquire MS~2 spectrum for each precursor ion current with a width of m/z=1.Because DI can provide desired measurement time, MS/MS~(ALL) is able to acquire MS~2 spectrum for each compound individually except for the compounds which share identical nominal molecular weight, even isomers.A total of 52 compounds were identified in Pien-Tze-Huang, including 16 saponins, 24 bile acids, 9 fatty acids, 2 saccharides, and 1 other compound.DI-MS/MS~(ALL) can simultaneously identify the compounds with different polarities in a short time, which is superior to LC-MS.This study provides a powerful tool for the rapid chemome profiling of Chinese medicinal prescriptions.

A strategy to discover lead chemome from traditional Chinese medicines based on natural chromatogram-effect correlation (NCEC) and natural structure-effect correlation (NSEC): Mahonia bealei and Mahonia fortunei as a case study.

Lead compound is an important concept for modern drug discovery. In this study, a new concept of lead chemome and an efficient strategy to discover lead chemome were proposed. Compared with the concept of lead compound, lead chemome can provide not only the starting point for drug development, but also the direction for structure optimization. Two traditional Chinese medicines of Mahonia bealei and Mahonia fortunei were used as examples to illustrate the strategy. Based on natural chromatogram-effect correlation (NCEC), berberine, palmatine and jatrorrhizine were discovered as acetylcholinesterase (AchE) inhibitors. Taking the three compounds as template molecules, a lead chemome consisting of 10 structurally related natural compounds were generated through natural structure-effect correlation (NSEC). In the lead chemome, the IC50 values of jatrorrhizine, berberine, coptisine, palmatine and epiberberine are at nanomolar level, which are comparable to a widely used drug of galantamine. Pharmacophore modeling shows that the positive ionizable group and aromatic rings are important substructures for AchE inhibition. Molecular docking further shows that pi-cation interaction and pi-pi stacking are critical for compounds to maintain nanomolar IC50 values. The structure-activity information is helpful for drug design and structure optimization. This work also expanded the traditional understanding of "stem is the medicinal part of Mahonia bealei and Mahonia fortunei". Actually, all parts except the leaf of Mahonia bealei exhibited potent AchE-inhibitory activity. This study provides not only a strategy to discover lead chemome for modern drug development, but also a reference for the application of different parts of medicinal plants.

Direct infusion-tandem mass spectrometry combining with data mining strategies enables rapid chemome characterization of medicinal plants: A case study of Polygala tenuifolia.

Data-independent MS2 spectrum acquisition after fragmenting the precursor ion cohort with 1 Da bin, termed as MS/MSALL ®, offers an opportunity to achieve rapid chemome characterization when being coupled with direct infusion (DI). Some post-acquisition data processing strategies, such as mass defect filtering (MDF), diagnostic fragment ion filtering (DFIF), and neutral loss filtering (NLF), facilitate data extraction from massive dataset, and moreover, molecular weight (MW) imprinting allows rapid capturing those reported components. Here, DI-MS/MSALL ® was employed to acquire cubic spectral dataset, and the strategies such as MW imprinting, MDF, DFIF, and NLF, were subsequently applied to filter the structural information. The integrated pipeline was utilized for the chemome characterization of Polygala tenuifolia, a famous edible medicinal plant. To aid information filtering, an in-house chemical library was built by comprehensively collecting structural information from some available databases. A single analytical run was completed within 5 min. For MS1 spectrum processing, MW imprinting was firstly applied to capture the compounds in the chemical library, and "five-point" MDF frames were employed to pursue saponins, oligosaccharide esters, and xanthones. Regarding MS2 spectral plot, DFIF and NLF were deployed to search information-of-interest. Structural identification was accomplished by carefully correlating precursor ions and MS2 spectra, applying the well-defined mass cracking rules, and referring to literature information as well as available databases. A total of 109 compounds, mainly saponins (40 ones), oligosaccharide esters (29 ones), and xanthones (19 ones), were captured and structurally annotated. MS1 spectra were also implemented for chemome comparison between Polygala tenuifolia and several similar plants belonging to Polygala genus, resulting in the observation of significant inter- and intra-species differences. Above all, DI-MS/MSALL ® is a promising choice for high-throughput chemome profiling of, but not limited to, medicinal plants, in particular when being integrated with post-acquisition data processing strategies.

[Chemome profiling of Pien-Tze-Huang by online pressurized liquid extraction-ultra-high performance liquid chromatography-ion trap-time-of-flight mass spectrometry].

Pien-Tze-Huang is one of the most famous traditional Chinese medicine prescriptions and consists of several precious medicinal materials, such as Notoginseng Radix et Rhizoma, Bovis Calculus, Snake Gall, and Moschus. However, its formula has not been completely revealed. It is mainly applied for the treatment of acute and chronic viral hepatitis, carbuncle, and boils caused by blood stasis, unknown swelling, bruises, and various inflammation disorders. The chemical composition of Pien-Tze-Huang is extremely complicated. Thus far, extensive attention has been paid to the principal chemical families in Pien-Tze-Huang, such as ginsenosides, bile acids, and muscone derivatives. Comprehensive chemical profiling, although of immense importance for systematic quality control, has not been achieved. Therefore, we configured a platform, namely online pressurized liquid extraction-ultra-high-performance liquid chromatography-ion trap-time-of-flight mass spectrometry (online PLE-UHPLC-IT-TOF-MS), to characterize the chemical profile of Pien-Tze-Huang in detail as well as to conduct source attribution, aiming to clarify the chemome of Pien-Tze-Huang and to provide a reliable method for quality assessment. A sub-microgram amount of Pien-Tze-Huang powder (0.3 mg) was placed in a hollow guard column, which was subsequently filled with clear silica gel. Filter membranes were used to seal the extraction vessel. The vessel was then placed in an adapted guard column holder and maintained in a thermal column oven (70 ℃). Metal tubing was used to connect the outlet of the guard column holder to the mass spectrometer. The extraction phase was maintained for 3 min by employing 0.1%(v/v) formic acid aqueous solution as the extraction solvent with a flow rate of 0.2 mL/min. Moreover, a six-port two-position electronic valve was introduced to automatically switch the system from extraction to elution phases. Within the elution phase, 0.1%(v/v) formic acid aqueous solution and acetonitrile composed the mobile phase, and the extracts were eluted with a gradient program. Because of the elevated temperature and pressure, the physical and chemical properties of water, especially polarity and solubility, were modified. Therefore, warm water could be an eligible green solvent to achieve wide polarity-spanned extraction. In addition, IT-TOF-MS was employed to acquire tandem mass spectrometry information. The mass fragmentation pathways of saponins and bile acids were carefully studied. Finally, according to authentic compounds, mass fragmentation pathways, reference information in the literature, and accessible databanks, a total of 73 signals were observed from Pien-Tze-Huang, of which 71 components were tentatively identified and assigned. Among them, 36 were from Notoginseng Radix et Rhizoma, 15 from Snake Gall, and 9 from Bovis Calculus, while the occurrences of the other 11 components were synergistically contributed by both Bovis Calculus and Snake Gall, through retrieving the in-house chemical database that was built by considering all accessible chemical information from Notoginseng Radix et Rhizoma, Bovis Calculus, Snake Gall, and Moschus. The other two compounds were assigned as unknown compounds. However, none of the components were assigned to Moschus because they mainly contained hydrophobic compounds, such as cycloketones, cholesterol, and sterols, among others, and it was difficult to detect them with the current measurement program. The extraction efficiency of online PLE was assessed by comparing it with the efficiency obtained from ultrasonication at the same time. According to base peak ion current chromatograms (BPCs) and mass spectrometry information, the efficiency of online PLE was greater than that of ultrasonic extraction, even through direct analysis. Online PLE-UHPLC-IT-TOF-MS is not only a tool fit for the concept of green analytical chemistry, but also a reliable analytical pipeline for the direct characterisation of other complicated matrixes. Above all, this study clarified the chemome of Pien-Tze-Huang and provided meaningful information for the quality control of this famous TCM prescription.

[Chemome profiling of Citri Reticulatae Pericarpium using UHPLC-IT-TOF-MS].

Citri Reticulatae Pericarpium(CRP, Chinese name: Chenpi) is one of the most famous edible traditional Chinese medicines(TCMs). CRP was first recorded as top grade TCM in Shennong Bencao Jing attributing to the benefits such as regulating Qi, tonifying spleen, eliminating dampness and eliminating phlegm, and has been widely utilized for the treatments of abdominal fullness and distention, vomiting and diarrhea, as well as phlegm cough. CRP is also widely popular as spice in food industry. Because of the wide cultivation, a number of brands that exhibit extensive price range can be found in the market, resulting in a great challenge for grading. Herein, an attempt was made to in-depth chemome profiling for the sake of providing meaningful information of the universal quality control of CRP. A new core-shell column packed with adamantylethyl substituted silica gel particles was deployed for chromatographic separations and IT-TOF-MS that is advantageous at providing abundant high resolution molecular and fragment ions was employed for qualitative detection. A total of 62 components were observed and 61 ones were structurally annotated according to proposing mass fragmentation patterns, matching with reference compounds and relevant databases, and the chemical families included flavone, limonin, etc. In particular, ten compounds bearing 3-hydroxy-3-methylglutarate substitute were detected from CRP for the first time. Above all, the chemical profile of CRP was characterized and the findings are meaningful for the in-depth quality assessment and efficacy material clarification of CRP.

[Direct chemome profiling of Boschniakia rossica using online pressurized extraction-ultrahigh-performance liquid chromatography-ion trap-time-of-flight mass spectrometry].

An online pressurized extraction-ultrahigh-performance liquid chromatography-ion trap-time-of-flight mass spectrometry (online PLE-UHPLC-IT-TOF-MS) platform was configured for the rapid and direct profiling of the chemical constituents of Boschniakia rossica (B. rossica). Notably, only a sub-microgram amount (0.5 mg) of B. rossica was placed in an empty guard column core, which was then filled up with normal-phase silica gel as an extraction vessel. The guard column core was placed inside a guard column holder in the column oven (70℃) and connected to the UHPLC-IT-TOF-MS system via metal tubing. The extraction phase was maintained for 3 min with 0.1% (v/v) formic acid as the extraction solvent. The extraction and elution phases for the entire measurement were segmented by a 6-port/2-channel electronic valve. In the elution phase, 0.1% (v/v) aqueous formic acid-acetonitrile containing 0.1% (v/v) formic acid was the mobile phase, and the components of interest that accumulated at the front of the column were eluted into the IT-TOF-MS system for detection. A total of 48 compounds were observed, and according to the literature, database, and mass fragmentation pathways, 45 compounds, including 10 phenylethanol glycosides, 14 iridoid glycosides, and 21 phenylalanine glycosides, were identified from B. rossica. This study provides reliable information regarding the chemical composition and quality evaluation of B. rossica. Moreover, it offers a promising analytical pipeline for the chemical composition characterization of traditional Chinese medicines.

Chemome profiling of Citri Reticulatae Pericarpium using UHPLC-IT-TOF-MS / 中国中药杂志

Citri Reticulatae Pericarpium(CRP, Chinese name: Chenpi) is one of the most famous edible traditional Chinese medicines(TCMs). CRP was first recorded as top grade TCM in Shennong Bencao Jing attributing to the benefits such as regulating Qi, tonifying spleen, eliminating dampness and eliminating phlegm, and has been widely utilized for the treatments of abdominal fullness and distention, vomiting and diarrhea, as well as phlegm cough. CRP is also widely popular as spice in food industry. Because of the wide cultivation, a number of brands that exhibit extensive price range can be found in the market, resulting in a great challenge for grading. Herein, an attempt was made to in-depth chemome profiling for the sake of providing meaningful information of the universal quality control of CRP. A new core-shell column packed with adamantylethyl substituted silica gel particles was deployed for chromatographic separations and IT-TOF-MS that is advantageous at providing abundant high resolution molecular and fragment ions was employed for qualitative detection. A total of 62 components were observed and 61 ones were structurally annotated according to proposing mass fragmentation patterns, matching with reference compounds and relevant databases, and the chemical families included flavone, limonin, etc. In particular, ten compounds bearing 3-hydroxy-3-methylglutarate substitute were detected from CRP for the first time. Above all, the chemical profile of CRP was characterized and the findings are meaningful for the in-depth quality assessment and efficacy material clarification of CRP.

From 1H NMR-based non-targeted to LC-MS-based targeted metabolomics strategy for in-depth chemome comparisons among four Cistanche species.

The great orthogonality between 1H NMR spectroscopy and LC-MS implies that their deployments in series could offer an opportunity to gain the qualified molecular markers via comparative metabolomics, and an attempt was made here to propose an integrated strategy namely "from 1H NMR-based non-targeted to LC-MS-based targeted metabolomics". In-depth chemome comparisons of Cistanche plants, such as C. deserticola, C. salsa, C. tubulosa, and C. sinensis, that possess dramatic economic and ecological benefits for the arid regions in the northwest China attributing to their dramatic medicinal and edible values, were employed to verify the applicability. 1H NMR-based non-targeted matabolomics acted as the survey experiment to find those signals offering decisive contributions towards the species discrimination, and the signals were translated to a set of putative identities, eighteen ones in total, through matching with authentic compounds and referring to some accessible databases. Afterwards, an advanced LC-MS platform assembling reversed phase liquid chromatography, hydrophilic interaction liquid chromatography, and tailored multiple reaction monitoring, was introduced to simultaneously quantify those eighteen potential markers in a single analytical run, because those candidates exhibited great polarity span as well as wide content range. Significant species differences occurred amongst their chemome patterns. Echinacoside, acteoside, betaine, mannitol, 6-deoxycatalpol, sucrose, and 8-epi-loganic acid were disclosed as the markers enabling the discrimination of those four species. The findings offered an alternative tool to differentiate Cistanche plants. More importantly, the strategy namely "from 1H NMR-based non-targeted to LC-MS-based targeted metabolomics" facilitates the pursuit of molecular markers among analogue plants, and thereby provides a promising choice for in-depth chemome comparison.

[Chemome comparison between cultivated and wild Cistanche tubulosa using ¹H-NMR spectroscopy].

As a famous tonic medicine, Cistanche tubulosa has been honored as "ginseng of the deserts" for centuries. Aiming to address the resource shortage as well as the wild resource protection towards this herbal medicine, wide cultivation has been achieved in the southern Xinjiang. Herein, in-depth chemome comparison was conducted between cultivated and wild plants using ¹H-NMR spectroscopy that is capable of comprehensively providing qualitative and quantitative information of given complicated matrices. Multivariate statistical analysis was employed to process the dataset as well as to consolidate that the cultivated plants are comparable to those wild ones in term of chemome. ¹H-NMR spectra of both wild and cultivated plants were acquired in parallel after extraction. Following direct overlaying, great similarity occurred between these two groups. A total of 28 compounds were tentatively identified by referring to authentic compounds together with those available databases, such as HMDB and BMRB. Following principal component analysis, none significant difference was observed between wild and cultivated groups. Above all, from the viewpoint of chemical profile, the cultivated plants were almost equal to the wild plants; therefore, the cultivated plants are able to take the load of wild plants in clinical usage. Moreover, ¹H-NMR spectroscopy is a promising tool for chemical profiling traditional Chinese medicines because of the potential towards simultaneously exhibiting both quantitative and qualitative information for complicated matrices.

Chemome comparison between cultivated and wild Cistanche tubulosa using ¹H-NMR spectroscopy / 中国中药杂志

As a famous tonic medicine, Cistanche tubulosa has been honored as "ginseng of the deserts" for centuries. Aiming to address the resource shortage as well as the wild resource protection towards this herbal medicine, wide cultivation has been achieved in the southern Xinjiang. Herein, in-depth chemome comparison was conducted between cultivated and wild plants using ¹H-NMR spectroscopy that is capable of comprehensively providing qualitative and quantitative information of given complicated matrices. Multivariate statistical analysis was employed to process the dataset as well as to consolidate that the cultivated plants are comparable to those wild ones in term of chemome. ¹H-NMR spectra of both wild and cultivated plants were acquired in parallel after extraction. Following direct overlaying, great similarity occurred between these two groups. A total of 28 compounds were tentatively identified by referring to authentic compounds together with those available databases, such as HMDB and BMRB. Following principal component analysis, none significant difference was observed between wild and cultivated groups. Above all, from the viewpoint of chemical profile, the cultivated plants were almost equal to the wild plants; therefore, the cultivated plants are able to take the load of wild plants in clinical usage. Moreover, ¹H-NMR spectroscopy is a promising tool for chemical profiling traditional Chinese medicines because of the potential towards simultaneously exhibiting both quantitative and qualitative information for complicated matrices.

An integrated strategy to quantitatively differentiate chemome between Cistanche deserticola and C. tubulosa using high performance liquid chromatography-hybrid triple quadrupole-linear ion trap mass spectrometry.

It is important to conduct large-scale detection, identification, and quantitation of metabolites in a given sample. Herein, a practical strategy was proposed to quantitatively compare the chemome between Cistanche deserticola (CD) and C. tubulosa (CT), which have been widely believed as the ideal edible and medicinal plants for conquering the deserts. The entire workflow was implemented on high performance liquid chromatography-hybrid triple quadrupole-linear ion trap mass spectrometer and consisted of three primary steps: (1) component detection and identification, various mass spectrometric approaches were applied to globally screen the chemical constituents, and structural elucidation was achieved by comparing with authentic compounds, analyzing MS(2) spectra, and referring to the literature along with accessible databases; (2) comprehensive relative quantitation, scheduled multiple reaction monitoring algorithm was introduced for relative quantitation of all detected ingredients; and (3) chemome comparison, the quantitative dataset was subjected for multivariate statistical analysis to carry out comparative study. A total of 513 metabolites were detected and relatively quantitated, and 379 ones were annotated. Betaine, Krebs cycle intermediates, phenylethanoid glycosides, and iridoids were picked out as the chemical markers being responsible for the discrimination of the chemical profiles between CD and CT. Above all, the quantitative chemome of CD and CT were exhaustively characterized and compared, which could advance their values concerning drug development, economics, and desertification control. The proposed strategy is expected as a reliable choice for widely targeted metabolomics of plants.