UHPLC-MS/MS analysis and protective effects on neurodegenerative diseases of phenolic compounds in different parts of Diospyros kaki L. cv. Mopan.

Persimmon (Diospyros kaki L. cv. Mopan.), an important commercial crop belonging to the genus of Diospyros in the Ebenaceae family, is rich in bioactive phenolic compounds. In this study, the phenolic compounds from fruits, leaves, and calyces of persimmon were qualitatively and quantitatively determined by UPLC-Q-Exactive-Orbitrap/MS and UPLC-QqQ-MS/MS, respectively. Furthermore, the role of phenolic extract from different parts of persimmon on neuroprotective activity in vitro, through against oxidative stress and anti-neuroinflammation effect was firstly evaluated. The results showed that 75 phenolic compounds, and 3 other kinds of compounds were identified, among which 44 of phenolic compounds were quantified from different parts of persimmon. It is the first time that epicatechin-epigallocatechin, catechin-epigallocatechin, catechin-epigallocatechin (A-type), and glycoside derivatives of laricitrin were identified in persimmon extract. The dominated phenolic compounds in three parts of persimmon were significantly different. All phenolic extracts from each part of persimmon showed strong neuroprotective activities against H2O2-induced oxidative stress in PC-12 cells and LPS-induced BV2 cells. The fruit extract presented the strongest activity, followed by calyx and leaf extract. The systematic knowledge on the phytochemical composition along with activity evaluation of different parts of persimmon could contribute to their targeted selection and development.

[Simultaneous determination of seven artemisinin-related compounds in Artemisia annua by UPLC-QQQ-MS/MS].

A variety of compounds in Artemisia annua were simultaneously determined to evaluate the quality of A. annua from multiple perspectives. A method based on ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS) was established for the simultaneous determination of seven compounds: amorpha-4,11-diene, artemisinic aldehyde, dihydroartemisinic acid, artemisinic acid, artemisinin B, artemisitene, and artemisinin, in A. annua. The content of the seven compounds in different tissues(roots, stems, leaves, and lateral branches) of A. annua were compared. The roots, stems, leaves, and lateral branches of four-month-old A. annua were collected and the content of seven artemisinin-related compounds in different tissues was determined. A multi-reaction monitoring(MRM) acquisition mode of UPLC-QQQ-MS/MS was used, with a positive ion mode of atmospheric pressure chemical ion source(APCI). Chromatographic separation was achieved on an Eclipse Plus RRHD C_(18) column(2.1 mm×50 mm, 1.8 µm). The gradient elution was performed with the mobile phase consisted of formic acid(0.1%)-ammonium formate(5 mmol·L~(-1))(A) and the methanol(B) gradient program of 0-8 min, 55%-100% B, 8-11 min, 100% B, and equilibrium for 3 min, the flow rate of 0.6 mL·min~(-1), the column temperature of 40 ℃, the injection volume of 5 µL, and the detection time of 8 min. Through methodological investigation, a method based on UPLC-QQQ-MS/MS was established for the simultaneous quantitative determination of seven representative compounds involved in the biosynthesis of artemisinin. The content of artemisinin in A. annua was higher than that of artemisinin B, and the content of artemisinin and dihydroartemisinic acid were high in all the tissues of A. annua. The content of the seven compounds varied considerably in different tissues, with the highest levels in the leaves and neither artemisinene nor artemisinic aldehyde was detected in the roots. In this study, a quantitative method based on UPLC-QQQ-MS/MS for the simultaneous determination of seven representative compounds involved in the biosynthesis of artemisinin was established, which was accurate, sensitive, and highly efficient, and can be used for determining the content of artemisinin-related compounds in A. annua, breeding new varieties, and controlling the quality of Chinese medicinal materials.

Identifying biomarkers of ginseng medicines with different natures on heart failure.

ETHNOPHARMACOLOGICAL RELEVANCE: The nature of Chinese medicine is a unique index to measure its efficacy. Generally, treating the hot syndrome with cold nature medicine and vice versa. Ginseng medicines, a renowned Chinese medicine known for its qi tonifying action, encompasses various herbal materials such as ginseng, red ginseng, and black ginseng (GS, RG, and BG, respectively), ginseng leaves (GL), and American ginseng (AG), which exhibited different natures, thought contained similar ginsenosides. This traditional effect of GS and RG "reinvigorate the pulse for relieving qi depletion". It is closely linked to anti-heart failure (HF), HF is a clinical manifestation of deficiency of "heart-qi". However, the elucidation of the mechanism underlying the anti-HF effects of ginseng medicines with different natures remains a significant challenge. AIM OF THE STUDY: To elucidate pharmacological mechanisms underlying the effect of ginseng medicines on HF, and to identify biomarkers associated with their various natures. Furthermore, it provides the basis for the different applications of ginseng medicines with various natures. MATERIALS AND METHODS: This study established a rat model of HF induced by isoproterenol (ISO) combined with a specific diet. Four representative hot/cold herbs were selected as compared references for the medicine natures. The divergent effects of these herbs on the HF model were investigated by analyzing RNA-seq data to identify genes expressed differentially. Additionally, pathways associated with medicine natures were obtained using KEGG. Furthermore, UPLC-QqQ-MS/MS, as well as ELISA, were used to measure indexes associated with the nervous system, energy metabolisms, and endocrinology systems, such as BNP, CK, IL-1, T3, T4, cAMP, cGMP, AD, adrenal hormones (DOC, CORT, and COR), progestogens (pregnenolone, P, 17-OH-PR, and 17-OH-P), androgens (DHEA, A4, and T), and estrogens hormones (E2). RESULTS: All ginseng medicines demonstrated varying levels of efficacy in alleviating HF and GS exhibited a significant protective effect on HF. The ginseng medicines with qi tonifying primarily achieve their effect by enhancing the levels of adrenal hormones (DOC, CORT, and COR), T4, elevation of cAMP/cGMP, and activation of AchE. Warm nature qi tonifying ginseng medicines increased the levels of 17-OH-PR and P while decreasing 17-OH-P and the ratio of E2/T. On the other hand, cold nature qi tonifying ginseng medicines decreased the levels of A4 and T while increasing the ratio of E2/T. CONCLUSION: Overall, the effects of warm nature ginseng medicines are stronger on HF compared to cold nature ginseng medicines. Our research firstly reported that the E2/T ratio, progestogens (17-OH-PR, 17-OH-P, and P), and androgens (A4 and T) have been identified as significant biomarkers for discerning the mechanism differences of ginseng medicines with differences natures in treatment of HF.

Chemometrics-based Chemical Analysis of Myrrh and Its Vinegar-processed Products by UPLC-MS/MS.

Myrrh is widely used in clinical practice but accompanied by obvious toxicity. According to traditional Chinese medicines theory, processing with vinegar can effectively reduce its toxicity. However, the detoxification processing technology of Myrrh and the corresponding mechanism have been unclear. The objective of this study is to systematically analyze the variation in chemical composition of raw Myrrh and its processed products using UPLC-Q-TOF-MS/MS coupled with chemometrics. A total of 75 compounds including 56 sesquiterpenoids, 2 diterpenoids, 15 triterpenoids and 2 other types were identified. Raw Myrrh and its processed products were divided into two major groups, and 14 chemical markers were selected out by principal component analysis and partial least square discriminant analysis. Additionally, the exact content of 5 representative chemical markers was determined to be significantly reduced after vinegar-processing by UPLC-QQQ-MS/MS. Moreover, multivariate statistical analysis and the quantitative results comprehensively indicated that the optimized processing method was processing at a ratio of 200 : 5 (Myrrh:vinegar). This research provides not only a reliable foundation for the study of Myrrh, but also a scientific reference for clinical use of this herb.

Physicochemical characteristics, polyphenols and antioxidant activities of Dimocarpus longan grown in different geographical locations.

Longan is widely consumed due to its high nutritional value. The growing area has substantial effect on nutrient component and secondary metabolism of fruits. The aim of this study was to analyze the differences in physicochemical characteristics, polyphenol profiles, and antioxidant activity of longan fruits grown in four regions of China. Two representative cultivars 'Shixia' and 'Chuliang' located in Chongqing, Guanxi, Zhanjiang and Hainan were collected and analyzed. The results showed that the fruit weights, edible rates, and total soluble solids were 5.63-12.57 g, 52.7-68.7% and 17.54-23.68%, respectively. The titratable acids, reducing sugars, vitamin C contents were 0.22-0.62%, 2.27-5.55% and 68.29-157.34 mg/100 g, respectively. Interestingly, contents of total polyphenols and antioxidant activities in longan pericarps from Chongqing were higher than those from low-latitude regions for two cultivars. In addition, 10 polyphenols were detected by UPLC-QqQ-MS/MS which showed that the content of polyphenols was much higher in longan pericarps than in pulps. The content of polyphenol profiles in longan was mainly influenced by its tissue distribution. Cultivar type may also affect the polyphenol profile of longan.

Discovery of the material basis of Jiuwei Xifeng granules using pharmaco-chemistry and pharmacokinetics.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiuwei Xifeng granules (JWXF) is primarily used for the treatment of Tourette syndrome (TS) with kidney-Yin deficiency and internal stirring of liver wind. However, few studies have focused on this issue. AIM OF THE STUDY: This study aimed to clarify chemical composition of JWXF using in vitro and in vivo pharmaco-chemistry and to provide a basis for the clinical use of JWXF using a strategy of pharmacokinetics. MATERIALS AND METHODS: In this study, the chemical constituents and in vivo metabolism of JWXF were evaluated using high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS/MS), and the time-dependent processes of the three main components in rats were detected using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQ-MS/MS). RESULTS: A total of 75 constituents were identified, including 22 alkaloids, 21 terpenes, 15 organic acids and their derivatives, and 17 other compounds. After administration, 12 compounds were identified in rat plasma, including 11 prototypes and one metabolite. Pharmacokinetic analysis showed that the effects of gentiopicroside, gastrodin, and sweroside in rats were dose-dependent when the dose of JWXF was 1-4 g/kg. They were rapidly absorbed and did not accumulate in the plasma after 7-day continuous intragastric administration. CONCLUSIONS: JWXF consists of 75 components, including alkaloids, terpenes, and organic acids. The three main compounds, gastrodin, gentiopicroside, and sweroside, undergo rapid absorption, elimination, and dose-dependent pharmacokinetics.

Simultaneous determination of Panax notoginseng total saponins in rabbit tears by UPLC-QqQ-MS/MS and its application to pharmacokinetic study.

Panax notoginseng total saponins (PNS), the main bioactive components of the radix and rhizome of Panax notoginseng (Burk.) F.H. Chen, could treat eye disorders. For the treatment of ocular diseases, eye drops are the first choice with the most common, economic and good compliance. So we proposed that PNS might be able to treat inflammatory ocular surface diseases by eye drops based on its anti-inflammatory and antioxidant activities. The short elimination half-life (t1/2) and rapid elimination of PNS after oral or intravenous administration may limit its application for eye disorders. Meanwhile, there is a lack of pharmacokinetic study on trace amount of tear samples with PNS eye drops. Therefore, a simple and sensitive ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS) method by multiple reaction monitoring (MRM) in positive ion mode was firstly developed and applied in the pharmacokinetic study of PNS in rabbit tears. Tears samples were prepared by protein precipitation using methanol. The linearity, limit of detection, limit of quantification, specificity, precision, repeatability, stability, recovery, and matrix effect have been investigated and passed their validation criteria. Compared with prior methods, this method has the advantages of rapid analysis, high sensitivity, simple sample preparation and less sample demands. The pharmacokinetic results indicated that PNS eye drops had a slower elimination and a longer t1/2 by topical ocular administration, which is expected to improve the success rate of eye drops in the treatment of anterior segment diseases. The ocular pharmacokinetics of PNS provides an experimental guidance and feasibility basis for in vivo effect verification of PNS eye drops in the future investigation.

Comparison of production of bioactive components in Zanthoxylum nitidum taproots from different regions in southern China.

Zanthoxylum nitidum (Roxb.) DC is a traditional Chinese herb from southern China and its 3-4-year old roots are used in medicine. However, there is a scarcity of studies on the differences in the content of different regions of the roots, as well as comprehensive evaluations of Z. nitidum from the main areas of production in China. This study used ultra performance liquid chromatography, triple quadrupole tandem mass spectrometry, HPLC, and Fourier-transform infrared spectroscopy to detect and identify the bioactive components from different parts and eight regions of 4-year-old roots of Z. nitidum. Our results revealed that the types and quantities of compounds extracted were similar in root bark and root wood, although the amount of alkaloids in the former was substantially higher. The contents of four alkaloids in samples from Guangdong were higher than those in Guangxi Province. Meanwhile, hierarchical cluster analysis and principal component analysis showed that the samples from different regions were effectively identified and evaluated based on alkaloids and other bioactive substances. Our findings have significant implications for Z. nitidum harvesting and usage, as well as for origin identification, quality evaluation, and sensible use of Z. nitidum resources.

Exploring Mechanism of Renshen Guben Oral Liquids in Treating Renal Fibrosis Based on Metabolomics and Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Renshen Guben oral liquids（RGOL） in treatment of mice with renal fibrosis based on metabolomics and network pharmacology. MethodC57BL/6 mice were randomly divided into control group， model group and RGOL group， 12 mice in each group. Except for the control group， mice in the other groups were induced into unilateral ureteral obstruction（UUO） model by UUO. After preparation of the model， an aqueous solution of 4.2 g·kg-1 extract powder was administered by gavage to RGOL group for 14 d， and an equal amount of distilled water was administered by gavage to the control and model groups. After the last administration on the 14th day， urine was collected and detected by ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry（UPLC-QQQ-MS/MS） with 0.1% formic acid aqueous solution as mobile phase A， and acetonitrile-isopropanol（70∶30） as mobile phase B for gradient elution（0-1 min， 5%B； 1-5 min， 5%-30%B； 5-9 min， 30%-50%B； 9-11 min， 50%-78%B； 11-13.5 min， 78%-95%B； 13.5-14 min， 95%-100%B； 14-16 min， 100%B； 16-16.1 min， 100%-5%B； 16.1-18 min， 5%B）， column temperature of 40 ℃， flow rate of 0.4 mL·min-1， electrospray ionization（ESI）， collection range of m/z 50-900. Through network pharmacology， the targets of components in RGOL and the targets of renal fibrosis were analyzed interactively， and the key components and key targets were screened by network topology analysis， and DAVID platform was used to predict the signaling pathways of RGOL for the treatment of renal fibrosis. ResultA total of 7 differential metabolites involving 8 metabolic pathways were identified in RGOL for the treatment of renal fibrosis. The network pharmacology revealed that 36 key components in RGOL were related to 7 differential metabolites， mainly ginsenosides， notoginsenosides and nucleotides. Based on the herbs-components-targets-pathways network， a total of 23 key targets related to the treatment of renal fibrosis by RGOL were highlighted， which together with the differential metabolites were involved in linoleic acid metabolism， arginine biosynthesis， tricarboxylic acid cycle（TCA）， arginine and proline metabolism and other pathways. ConclusionBased on metabolomics and network pharmacology， this study preliminarily identified 7 differential metabolites， 36 potential pharmacodynamic components and 23 key targets and 4 key pathways in RGOL for the treatment of renal fibrosis， providing an experimental basis for the clinical application and mechanism study of this preparation.

Simultaneous quantification of fourteen characteristic active compounds in Eucommia ulmoides Oliver and its tea product by ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS).

Various kinds of bioactive compounds contribute to versatile health-promoting properties of Eucommia ulmoides Oliver (E. ulmoides). In present study, we developed a UPLC-QqQ-MS/MS method for simultaneous quantification of fourteen characteristic active compounds, including 3 lignans, 4 iridoids, 3 flavonoids and 4 phenolics in E. ulmoides and its tea product for the first time. The running time of the method is 6.5 min. It has good linearity, sensitivity, precision, accuracy, and stability. Using this high-throughput method, the distributions of fourteen characteristic active compounds in E. ulmoides and its tea product were clarified. Also, it was found that E. ulmoides tea exhibited superiority in contents of chlorogenic acid as compared with natural resources. Overall, the study provided a rapid, reliable, and efficient analysis method, which could be applied for the quality evaluation of E. ulmoides natural resources and their relative products in the field of food and medicine.

Free, soluble conjugated and insoluble bonded phenolic acids in Keemun black tea: From UPLC-QQQ-MS/MS method development to chemical shifts monitoring during processing.

Phenolic acids, including benzoic acid and hydroxycinnamic acid derivatives, are the main compounds of black tea. An efficient and accurate analytical method to quantify ten phenolic acids was established and validated by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QQQ-MS/MS). The chemical shifts during the processing of Keemun black tea were analyzed and the phenolic acids were quantified. Compared with fresh tea leaves, after processing, the contents of free phenolic acids, including gallic acid, salicylic acid, p-coumaric acid, and ferulic acid, increased markedly. Still, the contents of soluble conjugated phenolic acids, including p-coumaroylquinic acid isomers and chlorogenic acid isomers, decreased. Furthermore, the total contents of lignin, and insoluble bonded phenolic acids decreased. The adduct of (-)-epigallocatechin gallate and 3-caffeoylquinic acid was detected in tea samples, and its content increased highly after fermentation. The developed and validated analytical method can be used to monitor the manufacturing process of black tea.

The localization of the alkaloids in Coptis chinensis rhizome by time-of-flight secondary ion mass spectrometry.

Background: Understanding the spatial distribution of active compounds can effectively evaluate the quality of decoction pieces of traditional Chinese medicine (TCM). Traditional methods are economical and practical but lack chemical information on the original distribution. Time-of-flight secondary ion mass spectrometry (TOF-SIMS), with the advantage of non-destructive detection of samples, can directly analyze the distribution of chemical compounds on the surface of various samples. Methods: In this study, TOF-SIMS image analysis technology was used to detect TCM for the first time. Taking Coptis rhizome (CR) as an example, a commonly used TCM, the distribution of the compounds in the cross-section of CR was studied. Meanwhile, ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLCQQQ-MS/MS) was used to verify the results of TOF-SIMS. Results: The distribution of nine active compounds: berberine, epiberberine, coptisine, palmatine, columbamine, jatrorrhizine, tetrahydricheilanthifolinium, and oxyberberine, was well imaged in the cross-section of CR by TOF-SIMS. The content of berberine and epiberberine was the highest; Palmatine distribution in the pith was more than that in other parts; Oxyberberine was mainly concentrated in the cork and xylem rays. Normalization analysis showed contents of these compounds increased along with the growth years. The result was consistent with UPLC-QQQ-MS/MS. Conclusion: The TOF-SIMS method can display the spatial distribution status of the active compounds of herbs, providing a basis for selecting the medicine site with non-destructive and fast detection.

Isolation and Identification of Arisaema Cum Bile Fermentation Strain and Preliminary Investigation on Its Compound Strain Fermentation / 中国实验方剂学杂志

ObjectiveIn order to solve the problem that the quality and stability of Arisaema Cum Bile in the fermentation process with hybrid bacteria were not easy to control， the microorganism in the fermentation process of Arisaema Cum Bile was isolated and identified， the dominant strains were screened and the fermentation process of Arisaema Cum Bile with compound bacteria was investigated. MethodThe submerged culture during the fermentation process of Arisaema Cum Bile was taken out for strain separation and purification. Bacteria and fungi multiphase identification and detection methods and automatic microbial analysis system were used to analyze and compare DNA sequences and identify microorganisms. The isolated and identified strains were respectively inoculated and fermented. After screening the dominant strains， a preliminary exploration of compound strain fermentation were carried out. The contents of index components in Arisaema Cum Bile fermented by compound strain and traditional Arisaema Cum Bile were compared by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry （UPLC-QqQ-MS/MS）. Mmobile phase was 0.1% formic acid acetonitrile solution （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-2 min， 35%-45%A； 2-10 min， 45%-48%A； 10-12 min， 48%-100%A； 12-12.01 min， 100%-35%A； 12.01-15 min， 35%-65%A）， the flow rate was set at 0.35 mL·min-1. The mass spectrographic analysis employed electrospray ionization （ESI）， negative ion acquisition mode and multiple reaction monitoring （MRM） scanning mode were adopted to collect information， the collection range was m/z 50-1 000. ResultEight microorganisms were isolated and identified from the submerged culture of Arisaema Cum Bile. Among them， Enterococcus sp. （anaerobic） and E. casseliflavus were selected as the dominant strains in the fermentation process. Compared with the traditional fermentation method， the contents of chenodeoxycholic acid， hyodeoxycholic acid and hyocholic acid in free cholic acid increased by 1.76， 0.06， 0.19 mg·g-1， respectively. In bound cholic acid， glycochenodeoxycholic acid， taurochenodeoxycholic acid， glycohyodeoxycholic acid， taurohyodeoxycholic acid， glycohyocholic acid， taurine porcine cholic acid decreased by 0.63， 0.23， 0.26， 0.16， 0.03， 0.04 mg·g-1， respectively. ConclusionArisaema Cum Bile with compound strain fermentation （Enterococcus sp. and E. casseliflavus） can be fermented more completely， the fermentation cycle can be shortened， and the quality and stability of products can be improved.

Development of a Pteris vittata L. compound database by widely targeted metabolomics profiling.

The objective of this work was the development of a detailed, extensive and reliable database of the metabolomes of P. vittata. Using an ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry system (UPLC-QqQ-MS/MS) and based on the knowledge of retention time and mass spectral characteristics of an in-house collection of authentic standards, we screened for the presence of a large collection of natural compounds. The database represents 359 authenticated metabolites, comprising 220 primary and 139 secondary metabolites (70 flavonoids, 16 phenylpropanoic acid derivatives, five coumarins, two stilbenoids, 14 benzoic acids, nine phenols, 20 alkaloids and three terpenoids). Comparison of the accumulation of these compounds in two tissues showed that the aerial parts were enriched in flavonols, whereas the subterranean parts were enriched in anthocyanins. The comprehensive database developed here will be beneficial in improving the understanding of the chemical basis of plant therapeutic profile using multivariate analysis, with a particular example of antioxidant activity.

[Qualitative and quantitative analysis of alkaloids in Eurycoma longifolia by HPLC-Q-TOF-MS combined with UPLC-QQQ-MS/MS].

A sensitive and efficient method was established and validated for qualitative and quantitative analysis of total alkaloids from the extract of Eurycoma longifolia by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS) combined with ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS). The HPLC-Q-TOF-MS conditions are as follows: Welch Ultimate XB-C_(18) column(4.6 mm×250 mm, 5 µm) with acetonitrile(containing 0.1% formic acid)-0.1% formic acid in water as mobile phase for gradient elution. The UPLC-QQQ-MS/MS conditions are as below: Agilent Eclipse Plus C_(18) column(2.1 mm×50 mm, 1.8 µm) with acetonitrile(containing 0.1% formic acid) and 0.1% formic acid in water as mobile phase for gradient elution. MS data were collected by electrospray ionization in positive ion mode. According to the comparison with reference standards and the accurate masses of molecules, a total of 17 alkaloids in E. longifolia extract were identified by HPLC-Q-TOF-MS. The UPLC-QQQ-MS/MS quantitative analysis result of 3 alkaloids showed that the linear ranges of them were good(r≥0.999 7) and the overall recoveries ranged from 108.8%-110.2%, with RSDs of 2.9%-5.3%. The method is accurate, reliable, and efficient, which can comprehensively reflect the constituents and content of alkaloids in E. longifolia. The result can serve as a reference for further elucidating its therapeutic material basis and quality control.

Qualitative and quantitative analysis of alkaloids in Eurycoma longifolia by HPLC-Q-TOF-MS combined with UPLC-QQQ-MS/MS / 中国中药杂志

A sensitive and efficient method was established and validated for qualitative and quantitative analysis of total alkaloids from the extract of Eurycoma longifolia by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS) combined with ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS). The HPLC-Q-TOF-MS conditions are as follows: Welch Ultimate XB-C_(18) column(4.6 mm×250 mm, 5 μm) with acetonitrile(containing 0.1% formic acid)-0.1% formic acid in water as mobile phase for gradient elution. The UPLC-QQQ-MS/MS conditions are as below: Agilent Eclipse Plus C_(18) column(2.1 mm×50 mm, 1.8 μm) with acetonitrile(containing 0.1% formic acid) and 0.1% formic acid in water as mobile phase for gradient elution. MS data were collected by electrospray ionization in positive ion mode. According to the comparison with reference standards and the accurate masses of molecules, a total of 17 alkaloids in E. longifolia extract were identified by HPLC-Q-TOF-MS. The UPLC-QQQ-MS/MS quantitative analysis result of 3 alkaloids showed that the linear ranges of them were good(r≥0.999 7) and the overall recoveries ranged from 108.8%-110.2%, with RSDs of 2.9%-5.3%. The method is accurate, reliable, and efficient, which can comprehensively reflect the constituents and content of alkaloids in E. longifolia. The result can serve as a reference for further elucidating its therapeutic material basis and quality control.

[Qualitative and quantitative analysis of quassinoid diterpenoids from Eurycoma longifolia by UPLC-Q-TOF-MS combined with UPLC-QQQ-MS/MS].

In this work, a sensitive and efficient method was established and validated for qualitative and quantitative analysis of major quassinoid diterpenoids constituents from the extract of Eurycoma longifolia by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and ultra-performance liquid chromatography coupled with a triple quadrupole mass spectrometry(UPLC-QQQ-MS/MS). The UPLC-Q-TOF-MS analysis was performed on an Agilent Eclipse Plus C_(18) RRHD(2.1 mm×100 mm,1.8 µm) column with acetonitrile and 0.1% formic acid water as mobile phase by gradient elution. The UPLC-QQQ-MS/MS analysis was performed on an Agilent Eclipse Plus C_(18) RRHD(2.1 mm×50 mm, 1.8 µm)column with acetonitrile and 0.1% formic acid water as mobile phase by gradient elution. The data was collected by electrospray ionization in positive mode. According to the contrast of the reference standards and the accurate masses of molecules, a total of 32 quassinoid diterpenoids in E. longifolia extract were identified by UPLC-Q-TOF-MS. For quantitative the linear range of 4 detected quassinoid diterpenoids were good(r≥0.999 6), and the overall recoveries ranged from 90.35% to 106.4%, with the RSD ranging from 1.8% to 3.6%. The method was accurate, reliable and efficient, and could comprehensively reflect the chemical constituents and content of E. longifolia, and could provide a reference for further elucidating its pharmacological basis and quality control.

Facile in-situ polymerization of polyaniline-functionalized melamine sponge preparation for mass spectrometric monitoring of perfluorooctanoic acid and perfluorooctane sulfonate from biological samples.

In this present work, a novel polyaniline-functionalized melamine sponge (PMs) was successfully prepared using a simple unstirred in-situ polymerization process. The PMs was characterized using a scanning electron microscope and contact angle measurements. Its adsorption performance was initially determined via dye adsorption assays, and the conditions affecting the synthesis including polymerization time, acidity, molar ratio, and number and sizes of raw melamine sponge were optimized. The PMs was then used as an efficient adsorbent for the development of a novel, low-cost method for the detection of two representative perfluorinated chemicals, perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS), using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS) with the internal standard method. To achieve the best extraction efficiency with this method, several variables were optimized, including adsorption time, pH value, the number of PMs, and desorption conditions. Calibration graphs showed a good linear degree at concentration ranging from 0.1 to 50 µg L-1 for PFOA and 0.01-10 µg L-1 for PFOS, with a coefficient of detection R2 = 0.998. The intra-day and inter-day relative standard deviations were found to range from 5.9% to 8.2% for PFOA, and 5.5% to 7.7% for PFOS. Under these optimized conditions, the method was successfully used to measure PFOA and PFOS content in real human serum and urine samples, with average spiked recoveries ranging from 79% and 91% for PFOA, and 81% to 87% for PFOS.

Effect of Steaming Time and Pressure on Contents of Six Ginsenosides in Ginseng Radix et Rhizoma by UPLC-QqQ-MS/MS / 中国实验方剂学杂志

Objective::The processing method of red ginseng was determined by comparing the effects of different steaming time and pressure on the total content of six ginsenosides. Method::The contents of ginsenoside Rg1, Re, Rf, Rb1, Rc and Rb2 were determined by ultra high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS). The Waters ACQUITY UPLC BEH C8 column (2.1 mm×100 mm, 1.7 μm) was used. The mobile phase was 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile solution (B) for gradient elution (0-4 min, 81%-79%A; 4-6.3 min, 79%-75%A; 6.3-6.5 min, 75%-71%A; 6.5-9.5 min, 71%A; 9.5-16.5 min, 71%-68.5%A; 16.5-16.6 min, 68.5%-60%A; 16.6-19 min, 60%-100%A). The flow rate was set at 0.4 mL·min-1 and the column temperature was set at 35 ℃. The mass spectrographic analysis employed electrospray ionization (ESI) and negative ion collection mode with capillary ionization voltage of 2.5 kV, desolvation temperature of 350 ℃, desolvation gas flow of 700 L·h-1 and cone gas flow of 50 L·h-1. Multiple reaction monitoring (MRM) mode was used to collect information, the collection range was m/z 100-1 500, detection was performed by MRM mode at m/z 799.59-637.49 for ginsenoside Rg1, m/z 945.54-475.79 for ginsenoside Re, m/z 799.59-475.49 for ginsenoside Rf, m/z 1 107.59-783.97 for ginsenoside Rb1, m/z 1 077.58-783.96 for ginsenoside Rc, m/z 1 077.75-191.19 for ginsenoside Rb2. Result::When the steaming time was 3 hours, the total mass fraction of six ginsenosides in each sample group was 7.099 8-16.768 5 mg·g-1, and the total amount of the six ginsenosides in atmospheric steaming was 2.5-12.6 times of that in pressurized steaming, which was obviously better than that in pressurized steaming. Conclusion::Under the conditions of this experiment, the best processing method of red ginseng is atmospheric steaming for 3 hours with fresh ginseng.

Solid phase extraction of pesticides from environmental waters using an MSU-1 mesoporous material and determination by UPLC-MS/MS.

This paper describes the synthesis of a silica based MSU-1 mesoporous solid and its application as sorbent in solid-phase extraction to pre-concentrate thirteen pesticides of low-high polarity (methomyl, cymoxanil, carbofuran, monolinuron, isoproturon, methidathion, methiocarb, malathion, phosalone, diazinon, penconazole, neburon and chlorotoluron) in ground and river water. The synthesis was based in an H-bonding interaction assembling (I0N0) between two non-ionic components (the inorganic silica surface, I0 and the polyethylene oxide template, N0) by adding tetraethoxysilane to the non-ionic surfactant Brij®100, the latter previously dissolved in HCl 1 M. 50 mL water samples adjusted at pH= 3.5 were passed, at a flow rate of 5 mL/min, through a home-made cartridge containing 50 mg of MSU-1 sorbent, pre-conditioned with 5 mL of ultrapure water; then, the cartridge was washed with 5 mL of ultrapure water. Following elution with 5 mL of acetonitrile, the pesticides were determined by ultra performance liquid chromatography coupled to triple quadrupole-mass spectrometry. Two selected reaction monitoring transitions were monitored per compound, the most intense one being used for quantification and the second one for confirmation. Three points were used for identification, as established in the Directive 96/23/EC for LC-MS/MS analysis, which deals with confirmatory methods for organic residues and contaminants listed in the Group B (veterinary drugs and contaminants). Medium matrix effect (|20%||50%|). Therefore, the standard addition methodology was applied by adding an adequate amount of the pesticide standard mixture to the final sample extract. All pesticides were quantified using this approach for practical reasons, thus avoiding two different calibrations. The method quantification limit (MQL) of pesticides was 0.01 µg/L for all of them, except for diazinon (0.1 µg/L). Recoveries of the target pesticides at MQL and 0.25 µg/L concentration levels in blank river water were in the range 70.1-113.5% and 86.7-107.3%, respectively, with RSDs lower than 16.3% and 15.7%, respectively. Four ground water samples and three river water samples, taken from Almería (Spain), were analyzed by the proposed method and only phosalone at a concentration level of 0.05 µg/L was found in one river water sample.