Exploring the ethnopharmacological significance of Cynara scolymus bracts: Integrating metabolomics, in-Vitro cytotoxic studies and network pharmacology for liver and breast anticancer activity assessment.

ETHNOPHARMACOLOGICAL RELEVANCE: Liver and breast cancers are the most dominant cancer types with high occurrence rates. Artichoke (Cynara scolymus L.) has been reputed for its traditional use in alleviating many liver and gallbladder ailments beside its anticancer activity against various types of cancer cells. AIM OF THE STUDY: To demonstrate detailed chemical matrices of the different plant parts and evaluate their cytotoxic activities aiming to unveil the relationship between these activities and the intrinsic metabolites using metabolomic studies, in-vitro experiments and network pharmacology. MATERIALS AND METHODS: Chemical profiling of extracts from the different plant parts (stems, leaves, bracts and receptacles) was performed using HPLC/QqQ/MS followed by unsupervised chemometric studies. In-vitro cytotoxic potentials of the extracts were evaluated on breast and liver cancer cell line then an OPLS study using linear regression was conducted. Consequently, a network pharmacology analysis on the most bioactive plant organ was applied. RESULTS: Unsupervised chemometric analysis revealed that kaempferol-3-O-α-L-rhamnopyranoside-7-O-ß-D-galacturonopyranoside, chrysoeriol-7-rutinoside and 1-caffeoylquinic acid were responsible for the segregation of the bract (CSB) segregated from the rest of the plant organs. Interestingly, CSB extract possessed the highest potential in-vitro cytotoxic activity against both liver and breast cancer cells (IC50 = 1.65 and 1.77 µg/mL). As expected, the aforementioned biomarkers were observed to be the discriminatory cytotoxic metabolites in the constructed supervised chemometric model. Network pharmacology analysis on CSB revealed 27 liver cancer-related metabolites of which, 1-caffeoylquinic acid was the most enriched one contributing to 13% of the total interactions. Furthermore, 38 target genes were involved, the most enriched of which were Aldo-keto reductase family 1 member B1 (AKR1B10) and interleukin-2 (IL-2). KEGG pathway analysis unveiled 23 significantly related pathways including metabolic pathways that possessed the lowest p-value (1.6E-5). CONCLUSION: The findings demonstrated that CSB is a significant source of cytotoxic metabolites against breast cancer and liver cancer cell lines, hence, drawing attention to the pharmaceutical and medicinal value of this negligible plant organ and paving the route for insightful research into its exact pharmacological cytotoxic mechanisms.

A novel standard-free detection of adulteration method for sildenafil derivatives in dietary supplements.

The rapid and accurate detection of illegal adulteration of chemical drugs into dietary supplements is a big challenge in the food chemistry field. Detection of compounds without a standard reference is even more difficult; however, this is a common situation. Here in this study, a novel "standard-free detection of adulteration" (SFDA) method was proposed and phosphodiesterase-5 inhibitor derivatives were used as an example to figure out the possibility and reliability of this SFDA method. After analysis by quadrupole coupled time of flight-tandem mass spectrometry detection and multivariable statistics, six common fragment ions were chosen to indicate whether adulteration was present or not, while 20 characteristic fragment ions indicated whether adulteration was by nitrogen-containing heterocycles or by anilines. Furthermore, the quantitative methods were conducted by high-performance liquid chromatography-tandem mass spectrometry. In a word, this strategy allows for a quick determination of dietary supplement adulteration without any need for standard materials, improving the efficacy of food safety testing.

Quantitative analysis of six sesquiterpene glycosides from Dendrobium nobile Lindl. under different growth conditions by high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry in MRM mode.

INTRODUCTION: The sesquiterpene glycosides (SGs) from Dendrobium nobile Lindl. have immunomodulatory effects. However, there are no studies on the growth conditions affecting its contents and quantitative analysis methods. OBJECTIVE: In the present study, a quantitative analysis method for six SGs from D. nobile was established. We explored which growth conditions could affect the contents of SGs, providing a basis for the cultivation and clinical application of D. nobile. METHODS: Firstly, based on the optimization of mass spectrometry parameters and extraction conditions for six SGs in D. nobile, a method for the determination of the contents of six SGs was established using high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (HPLC-QqQ-MS/MS) in multiple reaction monitoring (MRM) mode. Then, the methodology of the established method was validated. Secondly, the established method was applied to determine the contents of six SGs from 78 samples of D. nobile grown under different growth conditions. Finally, chemometrics analysis was employed to analyze the results and select optimal growth conditions for D. nobile. RESULTS: The results indicated significant variations in the contents of SGs from D. nobile grown under different growth conditions. The primary factors influencing SG contents included age, geographical origin, altitude, and epiphytic pattern. CONCLUSION: Therefore, the established method for determining SG contents from D. nobile is stable. In particular, the SG contents were relatively high in samples of 3-year-old D. nobile grown at an altitude of approximately 500 m on Danxia rocks in Chishui, Guizhou.

[Effects of tetramethylpyrazine on pharmacokinetics in plasma and brain dialysate and neuropathic pain in rat model of spared sciatic nerve injury].

This study aims to explore the effects of tetramethylpyrazine(TMP) on pharmacokinetics in plasma and brain dialysate and neuropathic pain in the rat model of partial sciatic nerve injury(SNI), and to investigate the correlation between the analgesic effect of TMP and its concentrations in the plasma and brain dialysate. Male SD rats were randomized into Sham, SNI, and SNI+TMP groups. Mechanical stimulation with von frey filaments and cold spray method were employed to evaluate the mechanical sensitivity and cold sensitivity of rats. Another two groups, Sham+TMP and SNI+TMP, were used to intubate the common jugular vein and implant microdialysis probes into the anterior cingulate gyrus(ACC), respectively.After intraperitoneal injection of TMP at a dose of 80 mg·kg~(-1), automatic blood collection and intracerebral microdialysis(perfusion rate of 1 µL·min~(-1)) systems were used to collect the blood and brain dialysate for 24 h. HSS T3 C_(18) reversed-phase chromatographic column(2.1 mm×50 mm, 2.5 µm) was used for liquid chromatographic separation. Gradient elution was carried out with the mobile phase of methanol-water(containing 0.005% formic acid) at a flow rate of 0.25 mL·min~(-1). Electrospray ion source was used for mass spectrometry, and the scanning mode was multi-reaction monitoring under the positive ion mode. The ion pairs for quantitative analysis were TMP m/z 137/122 and aspirin m/z 179/137, respectively. DAS 2.11 was used to calculate the pharmacokinetic parameters. The optimal time of TMP to exert the analgesia effect and inhibit cold pain sensitivity was 60 min after treatment. The TMP in the plasma and brain dialysate of SNI rats showed the T_(max) of 15 min and 30 min, the C_(max) of(2 866.43±135.39) and(1 462.14±197.38) µg·L~(-1), the AUC_(0-t) of(241 463.30±28 070.31) and(213 115.62±32 570.07) µg·min·L~(-1), the MRT_(0-t) of(353.13±47.73) and(172.16±12.72) min, and the CL_Z of 0.73 and 0.36 L·min·kg~(-1), respectively. The analgesic effect of TMP had a significant correlation with the blood drug concentration in the ACC, which indicated that this method was suitable for the detection of TMP in rat plasma and brain dialysate. The method is accurate, reliable, and sensitive and can realize the important value of the application of correlation analysis theory of "automatic blood collection-microdialysis/PK-PD" in the research on neuropathic pain.

Establishment of a multicomponent quality control method and the transfer characteristics of five markers from Qidongning Formula to rat tissues by HPLC-QQQ-MS/MS.

Introduction: Traditional Chinese medicine compound preparations have become an increasingly utilized strategy for tumour treatment. Qidongning Formula (QDN) is a kind of antitumour compound preparation used in hospitals, and it can inhibit the growth of lung cancer cells. However, due to the complexity of botanical drugs, the quality evaluation of QDN is inconsistent, affecting clinical efficacy and posing potential safety risks for clinical application. Additionally, tissue distribution is an integral part of the drug development process. Methods: To study the distribution characteristics of markers in compound preparations and rat tissues, a novel HPLC-QQQ-MS/MS quantitative analytical method was established to determine five markers in QDN simultaneously, and the method was verified. Results and discussion: The analytical results showed that the contents of salidroside (51.6 ± 5.75 µg/g), calycosin-7-O-ß-D-glucoside (94.2 ± 15.4 µg/g), specnuezhenide (371 ± 72.5 µg/g), formononetin (23.8 ± 5.39 µg/g), and polyphyllin I (87.7 ± 10.6 µg/g) were stable in different batches of QDN. After intragastric administration (13.5 g/kg) in rats for 1 h, four markers in the QDN, except polyphyllin I, were distributed in most tissues. QDN was distributed chiefly in the stomach and small intestine, followed by the liver or kidney. The study also found that specnuezhenide had the highest concentration in both QDN and rat tissues (102 ± 22.1 µg/g in the stomach), while formononetin had the highest transfer rate (0.351%) from QDN to rat intestines. The above research lays a quality research foundation for the antitumour application of QDN and provides a scientific reference for the quality control of Chinese medicine compound preparations.

Simultaneous determination of twenty-nine active compounds in fuzhengjiedu granules by HPLC-QQQ-MS/MS.

As an empirical medicine of traditional Chinese medicine, Fuzhengjiedu Granules have shown an effect against COVID-19 in clinical and inflammatory animal models. It is formulated with eight herbs, including Aconiti Lateralis Radix Praeparata, Zingiberis Rhizoma, Glycyrrhizae Radix Et Rhizoma, Lonicerae Japonicae Flos, Gleditsiae Spina, Fici Radix, Pogostemonis Herba, and Citri Reticulatae Pericarpium. This study established a high-performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS) method by simultaneously determining 29 active compounds in the granules with significant content differences. Separation by gradient elution using acetonitrile and water (0.1% formic acid) as mobile phases was performed on a Waters Acquilty UPLC T3 column (2.1 mm × 100 mm, 1.7 µm). A triple quadrupole mass spectrometer, operating in positive and negative ionization modes, was used for multiple reaction monitoring to detect the 29 compounds. All calibration curves showed good linear regression (r2 > 0.998). RSDs of precision, reproducibility, and stability of active compounds were all lower than 5.0%. The recovery rates were 95.4-104.9%, with RSDs< 5.0%. This method was successfully used to analyze the samples, and the results showed that 26 representative active components from 8 herbs were detected in the granules. While aconitine, mesaconitine, and hypaconitine were not detected, indicating that the existing samples were safe. The granules had the maximum and minimum content of hesperidin (27.3 ± 0.375 mg/g) and benzoylaconine (38.2 ± 0.759 ng/g). To conclude, a fast, accurate, sensitive, and reliable HPLC-QQQ-MS/MS method was established, which can simultaneously detect 29 active compounds that have a considerable difference in the content of Fuzhengjiedu Granules. This study can be used to control the quality and safety of Fuzhengjiedu Granules and provide a basis and guarantee for further experimental research and clinical application.

Natural variation on free, esterified, glycosylated and insoluble-bound phenolics of Rubus chingii Hu: Correlation between phenolic constituents and antioxidant activities.

This study sought to determine how the variation of soluble free phenolics (SFPs), soluble-bound (including esterified-bound phenolics (EBPs), glycosylated-bound phenolics (GBPs)) and insoluble-bound phenolics (ISBPs) affected the antioxidant properties (radical scavenging activities and oxidative damage caused by H2O2 in RAW264.7 cell) of Rubus chingii Hu during different maturity stages. Our result demonstrated that the young fruit stage could be considered as a proper harvesting period for Rubus chingii Hu to obtain high antioxidant activities. Distinct tendencies about abundance were found in four phenolic forms and followed the order: SFPs ≈ EBPs > GBPs ≈ IBPs, and antioxidant activities correlation: EBPs > GBPs > SFPs. Furthermore, total phenolic content, total flavonoid content and phenolics such as kaempferol-3-O-rutinoside, tiliroside and ellagic acid have a substantial correlation with antioxidant capacity. This study may supply a theoretical foundation for the utilization and exploitation of Rubus chingii Hu in different maturity periods.

Regional Variation of Chemical Characteristics in Young Marselan (Vitis vinifera L.) Red Wines from Five Regions of China.

The environmental conditions of wine regions determine the flavor characteristics of wine. The characterization of the chemical composition and sensory profiles of young Marselan wines from five wine-producing regions in China was investigated by gas chromatography-mass spectrometry (MS), high-performance liquid chromatography-triple-quadrupole MS/MS and descriptive analysis. Young Marselan wines can be successful discriminated based on concentrations of volatile compounds, but not phenolic compounds, by orthogonal partial least squares discriminant analysis according to regions. Compared to Jiaodong Peninsula (JDP) and Bohai Bay (BHB) regions, there were relatively lower average concentrations of varietal volatiles (mainly including ß-citronellol, geraniol, and (E)-ß-damasenone) and several fermentation aroma compounds (including isoamyl acetate, octanoic acid, decanoic acid, ethyl decanoate, etc.) but higher levels of acetic acid in Xinjiang (XJ), Loess Plateau (LP), and Huaizhuo Basin (HZB) regions, which were related to their characteristic environmental conditions. Marselan wines from HZB, LP, and XJ regions were characterized by lower L values and higher a and Cab values. Marselan wines from XJ were discriminated from the wines from other regions due to their higher concentrations of several flavonols. Sensory analysis indicated that Marselan wines from HZB region were characterized by relatively low intensities of floral and fruity aromas compared to other regions.

Application of HPLC-QQQ-MS/MS and New RP-HPLC-DAD System Utilizing the Chaotropic Effect for Determination of Nicotine and Its Major Metabolites Cotinine, and trans-3'-Hydroxycotinine in Human Plasma Samples.

The routine techniques currently applied for the determination of nicotine and its major metabolites, cotinine, and trans-3'-hydroxycotinine, in biological fluids, include spectrophotometric, immunoassays, and chromatographic techniques. The aim of this study was to develop, and compare two new chromatographic methods high-performance liquid chromatography coupled to triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS), and RP-HPLC enriched with chaotropic additives, which would allow reliable confirmation of tobacco smoke exposure in toxicological and epidemiological studies. The concentrations of analytes were determined in human plasma as the sample matrix. The methods were compared in terms of the linearity, accuracy, repeatability, detection and quantification limits (LOD and LOQ), and recovery. The obtained validation parameters met the ICH requirements for both proposed procedures. However, the limits of detection (LOD) were much better for HPLC-QQQ-MS/MS (0.07 ng mL-1 for trans-3'-hydroxcotinine; 0.02 ng mL-1 for cotinine; 0.04 ng mL-1 for nicotine) in comparison to the RP-HPLC-DAD enriched with chaotropic additives (1.47 ng mL-1 for trans-3'-hydroxcotinine; 1.59 ng mL-1 for cotinine; 1.50 ng mL-1 for nicotine). The extraction efficiency (%) was concentration-dependent and ranged between 96.66% and 99.39% for RP-HPLC-DAD and 76.8% to 96.4% for HPLC-QQQ-MS/MS. The usefulness of the elaborated analytical methods was checked on the example of the analysis of a blood sample taken from a tobacco smoker. The nicotine, cotinine, and trans-3'-hydroxycotinine contents in the smoker's plasma quantified by the RP-HPLC-DAD method differed from the values measured by the HPLC-QQQ-MS/MS. However, the relative errors of measurements were smaller than 10% (6.80%, 6.72%, 2.04% respectively).

Chemical characterisation and quantification of the major constituents in the Chinese herbal formula Jian-Pi-Yi-Shen pill by UPLC-Q-TOF-MS/MS and HPLC-QQQ-MS/MS.

INTRODUCTION: Jian-Pi-Yi-Shen pill (JPYSP) is a Chinese medicine formula developed for the treatment of anaemic patients with chronic kidney disease (CKD). OBJECTIVE: To investigate the chemical profile of JPYSP in the treatment of renal anaemia. METHODS: A method coupling ultra-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) was established to characterise the chemical constituents present in JPYSP. Subsequently, a high-performance liquid chromatography method coupled with triple-quadrupole tandem mass spectrometry (HPLC-QQQ-MS/MS) was developed to quantify the major constituents from the identified compounds related to the treatment of CKD and anaemia. RESULTS: A total of 71 compounds were tentatively identified from JPYSP, including saponins, flavonoids, sesquiterpenoids, coumarins, phenylpropanoids, anthranones, anthraquinones, tannins, phenolic acids and others. Amongst them, 12 compounds (i.e. astragaloside IV, calycosin, calycosin 7-O-glucoside, salvianolic acid A, rosmarinic acid, rhein, liquiritin, formononetin, atractylenolide I, dioscin, tanshinone IIA, and acteoside) were further quantified simultaneously by HPLC-QQQ-MS/MS. CONCLUSION: The newly developed approach is suitable for the chemical profiling analysis and quality control of JPYSP, and could lead to additional pharmacodynamic studies involving the components of JPYSP.

[Effects of gut microbiota on five absorbed components of Berberis kansuensis in rat serum by HPLC-QqQ-MS].

To elucidate the absorption and metabolism of alkaloids in Berberis kansuensis in vivo, a high performance liquid chromatography-triple quadrupole mass spectrometry(HPLC-QqQ-MS) method was developed to qualitatively and quantitatively analyze the absorption components in rat serum in multiple-reaction monitoring mode. The mobile phase consisted of 0.1% formic acid and acetonitrile with a gradient elution mode. In addition, to investigate the effects of gut microbiota on five absorbed components of B. kansuensis in rat serum, diabetic rat and pseudo germ-free diabetic rat models were established, and partial least squares discriminant analysis and One-way ANOVA were used to study the content differences of five components among different groups. In this study, a HPLC-QqQ-MS method for quantitative analysis of five components in rat serum after oral administration of B. kansuensis was established for the first time. It was found that there were differences in the five constituents in rat serum between different groups. By comparing the normal group with the diabetic model group, we found that the absorption and metabolism capacities of berberine and magnoflorine were different under the health and pathological conditions. It was also found that the serum levels of berberine, magnoflorine and jatrorrhizine in pseudo germ-free diabetic rats were significantly lower than those in diabetic rats, indicating that gut microbiota plays an important role in the metabolism of alkaloids of B. kansuensis in vivo. These results provide a good reference for clarifying the active ingredients of B. kansuensis in the treatment of diabetes.

Application of a multiclass screening method for veterinary drugs and pesticides using HPLC-QTOF-MS in egg samples.

A multi-class screening method for the determination of veterinary drugs and pesticides in eggs was developed by high performance liquid chromatography quadrupole-time-of-flight mass spectrometry (HPLC-QTOF-MS). A single-injection workflow was developed for qualitative analysis, and method validation was conducted in terms of retention time deviation (ΔRt), exact mass error (Δm), selectivity, specificity, precision and trueness. The characteristic fragment ions in information dependent acquisition (IDA) mode were summarized for retrospective and non-target screening. Five of 70 samples were found positive with sulfachloropyridazine, sulfamonomethoxine and fipronil sulfone. Compared with triple-quadrupole (QqQ) MS method, the obtained results showed that QTOF-MS gave comparable results to QqQ-MS with residue concentrations higher than 10 µg·kg-1, whereas QqQ-MS was more powerful at lower concentrations (<1 µg·kg-1). Moreover, the amount of fragment ions acquired by HPLC-QTOF-MS decreased substantially at trace levels, which may be a potential cause of reduced reliability of identification.

Method Development for Selected Bisphenols Analysis in Sweetened Condensed Milk from a Can and Breast Milk Samples by HPLC-DAD and HPLC-QqQ-MS: Comparison of Sorbents (Z-SEP, Z-SEP Plus, PSA, C18, Chitin and EMR-Lipid) for Clean-Up of QuEChERS Extract.

Background: Identification and quantitative determination of analytes released from the packaging material is undoubtedly a difficult and tricky task, requiring the chemical analyst to develop an individual approach to obtain reliable analytical information. Unfortunately, it is still challenging for scientists to determine bisphenols at trace or even ultra-trace levels in samples characterized by a very complex, and often variable, matrix composition. Objective: Optimization and application of QuEChERS/d-SPE coupled with HPLC-DAD (and LC-QqQ-MS) method for the simultaneous determination of bisphenols (A, S, F, B, BADGE and derivatives) in milk samples from a can and breast milk samples have been performed. Methods: Concerning the analysis of unconjugated analytes, after the thawing and shaking the sample (5 mL breast milk or 10 mL milk samples from a can), it was transferred into a 50 mL polypropylene centrifuge tube. For the analysis of the total amount of analytes, prior to the extraction with acetonitrile, a deconjugation step was implemented in a tube by adding to sample, the an Isotopically Labelled Internal Standard (IS) solution (50 ng/mL) and 1 mL of the enzymatic solution with the ß-Glucuronidase (3500 U/mL). The mix was homogenized and incubated for 16-18 h at 37 °C. Next, 10 mL of acetonitrile, and a QuEChERS salt packet (4 g anhydrous MgSO4, 1 g NaCl) were added. After shaking and centrifugation, the total acetonitrile layer was isolated in a polypropylene tube evaporate to dryness, and reconstitute in 1.2 mL acetonitrile. During d-SPE step the extract was transferred into a 15 mL polypropylene tube with Z-Sep and primary secondary amine (PSA). Next, shake the tube, store in fridge, and centrifuge for 15 min. The acetonitrile supernatant was obtained with a pipette and evaporated to dryness. Mixture MeOH: water (20:80, v/v) were added to the dry residue and the extract was reconstitute in 200 µL and analyzed by HPLC-DAD and HPLC-QqQ-MS equipment. Conclusion: Six different salts during d-SPE step were evaluated such as: zirconium dioxide-based sorbent (Z-Sep, Z-Sep Plus), primary secondary amine (PSA), octadecyl (C18), EMR-Lipid, Chitin and also their mixtures. Negligible matrix interference was observed for most of the analytes due to application of Z-Sep and PSA in dispersive-solid phase extraction clean-up step. Extraction of target analytes was performed using QuEChERS/d-SPE cleanup, and presents good performance for selected analytes with recoveries in the range of 15-103% and relative standard deviations (RSD) less than 10% in breast milk samples.

In vitro studies on the metabolism of saikogenins and the detection of their metabolites in authentic biosamples.

Saikosaponins are the representative bioactive ingredient in the Radix Bupleuri. Previous studies have reported that saikosaponins are prone to losing their glycones and being converted to corresponding prosaikogenins and saikogenins by intestinal bacteria. However, the microsomal cytochrome P450-mediated metabolism of these deglycosylated metabolites is still unknown. This research performed in vitro studies of five saikogenins in rat and human liver microsomes. High-performance liquid chromatography coupled with a hybrid ion trap and time-of-flight mass spectrometry (HPLC-IT/TOF-MS) was employed to identify the metabolites. To confirm the metabolites detected in vitro, plasma and feces obtained from rats administrating several saikogenins were also analyzed by high-performance liquid chromatography with triple-quadrupole mass spectrometry (HPLC-QqQ-MS). The in vitro metabolic stability of saikogenins was ranked as follows: SGF＞SGG＞SGE＞SGA＞SGD. A total of 71 metabolites generated by hydroxylation, carboxylation, and dehydrogenation, as well as combinations of these steps, were identified by accurate mass measurement and MSn fragmentation behavior. Among eight metabolic pathways, monohydroxylation or carboxylation was the major metabolic pathway both in vitro and in vivo. Analysis of in vivo biological samples suggested that analytical targets for saikogenins should be the compounds themselves and their oxidized metabolites. This research provides a basis for further studies of the in vivo metabolism of saikosaponins in humans.

[Simultaneous quantification and consistency analysis of 19 representative components in Longshengzhi Capsules by HPLC-QQQ/MS].

Longshengzhi capsule consisting of 12 herbs is widely used in clinically treating cerebral ischemia during recovery period.In this study,in order to investigate the consistency of different batches of Longshengzhi capsules,a high performance liquid chromatography coupled to triple quadrupole mass spectrometry method(HPLC-QQQ/MS) was developed for the determination of 19 representative components in Longshengzhi Capsules within 9 min. Methodology validation indicated this method was simple,rapid,accurate,highly sensitive and reproducible,and it could be used for the content determination of components in Longshengzhi Capsules. The consistency analysis results showed that paeoniflorin and calycosin-7-glucoside in Longshengzhi Capsules had the highest content; RSD value of total content of 19 compounds was 5. 2% and the RSD value of main compounds such as astragaloside and calycosin-7-glucoside was all less than 15%,reflecting good consistency among different batches. This study has provided a scientific method and basis for the quality control and consistency evaluation of Longshengzhi Capsules.

The study of the constituents and source of toxicants in poisonous honey.

A novel method for non-target screening of toxicants in poisonous honey was established in this study. Poisonous honey and nontoxic honey were contrastive detected using liquid chromatography quadrupole-time-of-flight mass spectrometry and analyzed by Mass Profiler Professional Software. 4 poisonous alkaloids were screened out and confirmed by comparison with reference compounds. In order to investigate the source of these poisonous alkaloids, 6 poisonous alkaloids, ubiquitous in Gelsemium elegan, from honey, honeybees, pollen in honeycomb and different organs of Gelsemium elegan, were quantified by liquid chromatography triple-quadrupole tandem mass spectrometry. The results showed that alkaloids composition characteristics in honey, honeybees, and pollen were similar to those in the flower and bud of Gelsemium elegan and significant different from those in leave, stem and root. This result demonstrated that poisonous alkaloids in honey were come from the gathering honey process. This strategy provided an efficient and rapid method for non-target screening of toxicants in food.

Comparison of α-glucosidase inhibitory effect and bioactive constituents of Anemarrhenae Rhizoma and Fibrous Roots.

Comprehensive utilization of medicinal plant resources is of great significance for sustainable development of traditional Chinese medicines. In the present study, the α-glucosidase inhibitory activities of the rhizome and fibrous root of Anemarrhena Asphodeloides Bunge, were compared detailedly, and a high performance liquid chromatography coupled with electrospray ionization tandem triple quadrupole mass spectrometry (HPLC-QQQ/MS) method was developed for simultaneous quantification of seven bioactive constituents including neomangiferin, mangiferin, isomangiferin, timosaponin BII, timosaponin B, timosaponin AIII, and timosaponin N in 40 batches of samples. The results demonstrated that fibrous root extracts had more potent α-glucosidase inhibitory activity than rhizome extracts. Mangiferin and isomangiferin were abundant in fibrous root, while the analyzed saponins were rich in rhizome. Based on the chemometrics methods including principal component analysis (PCA), orthogonal partial least square discriminant analysis (OPLS-DA), and partial least square (PLS), mangiferin and isomangiferin might be mainly responsible for α-glucosidase inhibitory activity of the genus. These findings indicate that the established HPLC-QQQ/MS method was proven to be useful and efficient for quality control of Anemarrhena materials, and fibrous root had the potential to be utilized as anti-diabetic medicinal resource.

Mycotoxins, pesticides and toxic metals in commercial spices and herbs.

A total of 300 samples representing six condiments (black pepper, basil, oregano, nutmeg, paprika, and thyme) were analysed for 11 mycotoxins, 134 pesticides and 4 heavy metals by ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry and inductively coupled plasma mass spectrometry. Mycotoxins were detected in 4%, 10% and 30% of all nutmeg, basil and thyme samples, respectively. The residues of 24 pesticides were detected in 59% of the analysed condiments. The maximum residue levels of pesticides were exceeded in 10% of oregano and 46% of thyme samples. A risk assessment of heavy metals was performed, indicating daily intake levels far below the tolerable intake levels.

Dynamic metabolic profile of Zhi-Zi-Da-Huang decoction in rat urine based on hybrid liquid chromatography-mass spectrometry coupled with solid phase extraction.

Zhi-Zi-Da-Huang decoction (ZZDHD) has been used for treatment of alcoholic liver disease in China for thousands of years. In order to reveal the dynamic biotransformation of the decoction in vivo, a high-throughout, sensitive and special method based on high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry (HPLC-DAD-TOF/MS) and high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-QqQ/MS) was developed and validated. 25 parent compounds and 28 metabolites were characterized, among which, two metabolites were found for the first time and tentatively identified by neutral-loss scan and product-ion scan. All the compounds were assigned to iridoids, flavones, anthraquinones, coumarin or p-coumaric acid, and their biotransformation pathways were found to involve glucuronidation, sulfation, reduction and ring cleavage. Glucuronidation occurred as a major metabolic pathway of genipin and flavanone and the conjugates could be detected almost during the whole sampling duration. To compounds such as anthraquinones, coumarin and p-coumaric acid, sulfation is the only transformation pathway and the metabolites were found at 0-12, 4-18, 4-48h respectively after administration. Reduction and/or ring cleavage of genipin glucuronide and naringin were also observed obviously. The phenomena that parts of parent compounds and metabolites were able to be detected even 48h after administration implied that the accumulating effect of these constituents in vivo would happen and the potential toxicity of the decoction might appear if multiple dosing is adopted. The strategy used in this paper was proved helpful to offer important information for the clinical safe use of ZZDHD.

Qualitative and quantitative analysis of the chemical constituents in Mahuang-Fuzi-Xixin decoction based on high performance liquid chromatography combined with time-of-flight mass spectrometry and triple quadrupole mass spectrometers.

High-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF/MS) and high-performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-QQQ/MS/MS) were utilized to clarify the chemical constituents of Mahuang-Fuzi-Xixin Decoction. There are 52 compounds, including alkaloids, amino acids and organic acids were identified or tentatively characterized by their characteristic high resolution mass data by HPLC-QQQ/MS/MS. In the subsequent quantitative analysis, 10 constituents, including methyl ephedrine, aconine, songrine, fuziline, neoline, talatisamine, chasmanine, benzoylmesaconine, benzoylaconine and benzoylhypaconine were simultaneously determined by HPLC-QQQ/MS/MS with multiple reaction monitoring mode. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.9992). The relative standard deviations (RSD) of inter- and intra-day precisions were <3%. This method was also validated by repeatability, stability and recovery with RSD <3% respectively. A highly sensitive and efficient method was established for chemical constituents studying, including identification and quantification of Mahuang-Fuzi-Xixin decoction.