Linear Ion-trap Tandem Mass Spectrometry Research Articles

Quality evaluation of Lysimachiae Herba from different habitats based on simultaneous determination of multiple bioactive constituents combined with multivariate statistical analysis

A method based on ultra-high performance liquid chromatography coupled with triple quadrupole linear ion trap-tandem mass spectrometry(UHPLC-QTRAP-MS/MS) was developed for the simultaneous determination of 41 bioactive constituents of flavonoids, organic acids, nucleosides, and amino acids in Lysimachiae Herba. The content of multiple bioactive constituents was compared among the samples from different habitats. The chromatographic separation was performed in a Waters XBridge®C_(18) column(4.6 mm×100 mm, 3.5 μm) at 30 ℃. The gradient elution was performed with 0.4% methanol(A)-formic acid water(B) as the mobile phase at a flow rate of 0.8 mL·min~(-1), and the multiple-reaction monitoring(MRM) mode was adopted. According to the content of 41 constituents, hierarchical cluster analysis(HCA), orthogonal partial least squares discriminant analysis(OPLS-DA), and gray relational analysis(GRA) were perfomed to comprehensively evaluate the samples from different habitats. The results showed that the 41 constituents exhibited good linear relationship within the tested concentration ranges, with the correlation coefficients(r) greater than 0.999 4. The method featured good precision, repeatability, and stability with the relative standard deviations(RSDs) less than 5.0%. The average recoveries of the 41 constituents ranged from 98.06% to 101.9%, with the RSDs of 0.62%-4.6%. HCA and OPLS-DA separated 48 batches of Lysimachiae Herba samples from different habitats into three categories: the producing areas in Sichuan and Chongqing, the producing areas in Jiangsu, Zhejiang, and Jiangxi, and the producing areas in Guizhou. The content of 41 constituents varied among the Lysimachiae Herba samples from different habitats. The GRA results revealed that the Lysimachiae Herba sample from Nanchong City, Sichuan Province had the best comprehensive quality. The method developed in this study was accurate and reliable and thus can be used for comprehensive evaluation of Lysimachiae Herba quality and provide basic information for the selection of habitats.

Pharmacokinetics and tissue distribution of 12 major active components in normal and chronic gastritis rats after oral administration of Weikangling capsules

Ethnopharmacological relevanceWeikangling Capsules (WKLCs) have been used in the clinic for the treatment of gastrointestinal disorders for more than 30 years. However, the pharmacokinetic characteristics and tissue distribution of its major bioactive components in rats under different physiological and pathological conditions are unclear. Aim of the studyIn this study, we aimed to clarify the differences in pharmacokinetic parameters and tissue distribution of the major active components in WKLCs under physiological and pathological states. Materials and methodNormal and ethanol-induced chronic gastritis rats received 2.16 g/kg WKLCs by gavage, and urine, feces, plasma, and tissue (heart, liver, spleen, lung, kidney, stomach, and small intestine) samples were obtained. The active components in urine, feces and plasma were detected by ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). A rapid and sensitive analytical method, ultra-high-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UHPLC-QTRAP-MS/MS), was established and validated to clarify and compare the pharmacokinetics and tissue distribution of the major active components in normal and chronic gastritis rats. ResultsA total of 36 chemical components in the feces, urine, and plasma of chronic gastritis rats were identified by UHPLC-Q-TOF-MS/MS. Among them, 20 were the prototype components of WKLCs, and 16 were metabolites. The pharmacokinetic characteristics and tissue distribution of 12 prototype components were successfully analyzed by UHPLC-QTRAP-MS/MS. The pharmacokinetic results showed that the Cmax, AUC0-t, and AUC0-∞ of paeoniflorin, glycyrrhizic acid, and glycyrrhetinic acid were distinctly higher than those of the other components in normal and chronic gastritis rats. Compared to normal rats, the Cmax, AUC0-t, and AUC0-∞ of albiflorin, liquiritin apioside, liquiritin, isoliquiritin, ononin, isoliquiritigenin, dactylorhin A, and glycyrrhizic acid were significantly increased in chronic gastritis rats (P < 0.05), while the Cmax, AUC0-t and AUC0-∞ of militarine and liquiritigenin had significantly lower decreases in chronic gastritis rats (P < 0.05). The results of the tissue distribution showed that the 12 components were widely distributed in the heart, liver, spleen, lung, kidney, stomach, and small intestine of rats, of which the liver, kidney, stomach, and small intestine were the main accumulative organs. Compared with normal rats, the concentrations of 12 components in the liver, kidney, stomach, and small intestine of chronic gastritis rats were widely higher than those of normal rats at the same time points. ConclusionThe pharmacokinetic characteristics and tissue distribution of 12 active components of WKLCs were comprehensively characterized and elucidated in normal and chronic gastritis rats. These findings laid a solid foundation for revealing the pharmacodynamic material basis of WKLCs in treating gastrointestinal disorders.

Simultaneous quantification of multiple active components of the ginkgo ketoester tablet when combined with donepezil in four biological matrices from an Alzheimer’s animal model: Evaluation of drug distribution patterns in vivo

Ginkgo ketoester tablets (GT) and donepezil are clinically used in combination to treat Alzheimer’s disease (AD). This study aimed to investigate the anti-dementia effects of two drugs alone and in combination by monitoring the distribution patterns of their active components in different biological matrices. A practical analytical method was developed for simultaneously quantifying 38 active compounds in different matrices at trace levels using ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UHPLC-MS2). Under optimized conditions, good chromatographic separation of the 38 target compounds was achieved within 14 min, with acceptable linearity, limits of detections and quantifications, precision, stability, and extraction recovery and matrix effects. The developed analytical method quantified the 38 active components in the brain, plasma, intestine, and intestinal contents of AD mice treated with GT alone, donepezil alone, and a combination of the two drugs. The result showed that the combination group had enhanced anti-dementia active compounds and reduced levels of toxic compounds entering the targeted tissues. These findings support the combined use of GT and donepezil for increasing anti-dementia treatment efficiency and reducing toxicity. The proposed UHPLC-MS2 strategy could be used to monitor the levels of active compounds from different biological matrices in future studies.

Quantitative analysis of nutrients for nucleosides, nucleobases, and amino acids hidden behind five distinct regions-derived Poria cocos using ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry.

Poria cocos is an edible fungus used as a health product and traditional Chinese medicinal preparation. Nevertheless, little is known about its nutrients. In this study, ultra-high performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry was conducted to quantify nucleosides, nucleobases, and amino acids in 32 batches of Poria cocos samples collected from Anhui, Sichuan, Hubei, Hunan, and Guizhou. Subsequently, the linearity, precision, repeatability, stability, and recovery of our methods were validated. Samples from different regions were clearly separated by partial least squares discriminant analysis and cluster analysis. Our results suggested that Poria cocos samples from different geographical environments differed in nucleosides, nucleobases, and amino acids. The plot of variable importance for projection disclosed differential compositions of L-Leucine, Uridine, L-Asparagine, L-Glutamine, L-phenylalanine, L-Ornithine monohydrochloride, L-Hydroxyproline, Taurine, and Inosine in Poria cocos from five regions. We found the highest content of total analytes, total amino acids, and total non-essential amino acids in Poria cocos from Anhui, total essential amino acids in the Sichuan samples, and total nucleosides in the Hunan samples. Overall, we determined the content of Poria cocos-derived nucleosides, nucleobases, and amino acids, providing the foothold for further chemical mining and use of Poria cocos.

Simultaneous Determination of 32 Pyrrolizidine Alkaloids in Two Traditional Chinese Medicine Preparations by UPLC-MS/MS.

Pyrrolizidine alkaloids (PAs) constitute a class of phytotoxin which demonstrates strong hepatotoxicity. In China, many plants containing PAs are used as traditional medicines or medicinal preparations, which could harm human health and safety. Xiaoyao Tablet (XYT) is an antidepressant drug registered in the European Union (EU), Compound Danshen Dropping Pills (CDDP) is a commonly used drug for coronary heart disease, and phase III clinical study is ongoing in the United States. The purpose of this study is to provide data to support the use of Chinese medicine preparations internationally and to establish analytical methods for 32 PAs in XYT and CDDP. The extraction parameters that were optimized include solid-phase extraction (SPE) cartridge, extraction method, and extraction solvent. Then ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-traptandem mass spectrometry (UPLC-MS/MS) was developed to effectively and efficiently quantify the 32 PAs of the XYT and CDDP. The analytical methods for XYT and CDDP were verified respectively. For XYT, the analytical method for 32 PAs was linear, and the correlation coefficient r was greater than 0.994; the recovery (REC%) at 10–2000 μg/kg was 73.3%–118.5%, and the relative standard deviation (RSD%) was 2.1%–15.4%. The CDDP REC% was 71.8%–112.0%, and the RSD% was 2.0%–17.1%. This study provides technical and data support for the registration of Chinese patented medicines in the EU, controls quality and ensures safety, and is committed to the internationalization and standardization of Chinese patented medicines.

Simultaneous Determination and Health Risk Assessment of Four High Detection Rate Pesticide Residues in Pu’er Tea from Yunnan, China

Four pesticides with a high detection rate in Pu’er tea have been determined by a QuEChERS (quick, easy, cheap, effective, rugged, safe) method with multiwalled carbon nanotubes (MWCNTs), and combined ultrahigh-performance liquid chromatography–triple quadrupole linear ion trap-tandem mass spectrometry (UHPLC-QTRAP-MS/MS). MWCNs have been compared with other common purification materials, and found to be superior. The matrix effect was systematically studied, and the results show that the MWCNs can quickly and effectively reduce matrix interference values, which were in the range from −17.8 to 13.8. The coefficients (R2) were greater than 0.99, with the limit of quantification ranging from 0.1 to 0.5 μg/kg, and the recovery rate ranging from 74.8% to 105.0%, while the relative standard deviation (RSD) ranged from 3.9% to 6.6%. A total of 300 samples, taken from three areas in which Yunnan Pu’er tea was most commonly produced, tested for four pesticides. The results show that the detection rate of tolfenpyrad in Pu’er tea was 35.7%, which is higher than other pesticides, and the lowest was indoxacarb, with 5.2%. The residual concentrations of chlorpyrifos, triazophos, tolfenpyrad and indoxacarb ranged from 1.10 to 5.28, 0.014 to 0.103, 1.02 to 51.8, and 1.07 to 4.89 mg/kg, respectively. By comparing with China’s pesticide residue limits in tea (GB 2763-2021), the over standard rates of chlorpyrifos, tolfenpyrad, and indoxacarb were 4.35%, 0.87% and 0%, respectively. The risk assessment result obtained with the hazard quotient (HQ) method shows that the HQ of the four pesticides was far less than one, indicating that the risk is considered acceptable for the four pesticides in Pu’er tea. The largest HQ was found for tolfenpyrad, 0.0135, and the smallest was found for indoxacarb, 0.000757, but more attention should be paid to tolfenpyrad in daily diets in the future, because its detection rate, and residual and residual median were all relatively high.

Discovery of Quality Markers of Nucleobases, Nucleosides, Nucleotides and Amino Acids for Chrysanthemi Flos From Different Geographical Origins Using UPLC-MS/MS Combined With Multivariate Statistical Analysis.

Nucleobases, nucleosides, nucleotides and amino acids, as crucial nutrient compositions, play essential roles in determining the flavor, function and quality of Chrysanthemi Flos. The quality of Chrysanthemi Flos from different geographical origins is uneven, but there have been no reports about the screening of their quality markers based on nutritional ingredients. Here, we developed a comprehensive strategy integrating ultra performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UPLC–MS/MS) and multivariate statistical analysis to explore quality markers of Chrysanthemi Flos from different geographical origins and conduct quality evaluation and discrimination of them. Firstly, a fast, sensitive, and reliable UPLC–MS/MS method was established for simultaneous quantification 28 nucleobases, nucleosides, nucleotides and amino acids of Chrysanthemi Flos from nine different regions in China. The results demonstrated that Chrysanthemi Flos from nine different cultivation regions were rich in the above 28 nutritional contents and their contents were obvious different; however, correlation analysis showed that altitude was not the main factor for these differences, which required further investigation. Subsequently, eight crucial quality markers for nine different geographical origins of Chrysanthemi Flos, namely, 2′-deoxyadenosine, guanosine, adenosine 3′,5′-cyclic phosphate (cAMP), guanosine 3′,5′-cyclic monophosphate (cGMP), arginine, proline, glutamate and tryptophan, were screened for the first time using partial least squares discriminant analysis (PLS-DA) and the plot of variable importance for projection (VIP). Moreover, a hierarchical clustering analysis heat map was employed to intuitively clarify the distribution of eight quality markers in the nine different regions of Chrysanthemi Flos. Finally, based on the contents of selected eight quality markers, support vector machines (SVM) model was established to predict the geographical origins of Chrysanthemi Flos, which yielded excellent prediction performance with an average prediction accuracy of 100%. Taken together, the proposed strategy was suitable to discover the quality markers of Chrysanthemi Flos and could be used to discriminate its geographical origin.

An integrated strategy of MS-network-based offline 2DLC-QTOF-MS/MS coupled with UHPLC-QTRAP®-MS/MS for the characterization and quantification of the non-polysaccharides in Sijunzi decoction.

Sijunzi decoction (SJZD), a classic recipe in traditional Chinese medicine (TCM), has been applied for the clinical treatment of gastrointestinal diseases. While there are reports on pharmaceutical substances of SJZD focusing on its polysaccharides, the composition of non-polysaccharides (NPSs) has not yet been holistically clarified. In the current study, offline two-dimensional liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (2DLC-QTOF-MS/MS) was used for comprehensive NPS chemical profiling of SJZD. In addition, the MS-network of SJZD was proposed, which led to the construction of a larger in-house chemical library and accelerated qualitative processing. Four hundred forty-nine components, among which 6 were potentially novel, and 32 were confirmed by standard substances, were identified or tentatively assigned. Furthermore, based on good method validation, 19 representative components were simultaneously quantified by ultra-high-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UHPLC-QTRAP®-MS/MS). They were selected for quantification on the account of their bioactive reports on in vivo or in vitro activities, the peak intensity in the mass spectrum, and characteristic structures, which have the potential to be qualitative or quantitative markers of SJZD. The present work furthers understanding of the pharmacological effects and action mechanism of NPSs in SJZD, and provides a useful analytical approach for complex composition research of TCMs.Graphical abstract Supplementary InformationThe online version contains supplementary material available at 10.1007/s00216-021-03302-x.

HILIC-UHPLC-QTRAP®/MS2 quantification of 15 neurotransmitters of the combination of donepezil and ginkgo ketoester tablet in different biological matrices from dementia mice: Application to study the synergistic effect of the two drugs

Neurotransmitters (NTs) were signaling molecules which played a key role in the communication of nerve cells. Simultaneous monitoring of NTs possessed important scientific significance for dementia related pharmacology, pathology and drug study. This study developed an effective method for analysis of 15 underivatized NTs on hydrophilic interaction ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (HILIC-UHPLC-QTRAP®/MS2). Under the optimized chromatographic conditions, good separation of 15 NTs was achieved within 5.0 min. The developed HILIC-UHPLC-QTRAP®/MS2 was validated by matrix effects, linearity, limits of detections and quantifications, precision, stability and recovery. Finally, the validated analytical method was applied to quantify 15 NTs in brain, plasma, intestinal and intestinal contents from normal, dementia model and the combination of ginkgo ketoester tablet (GT) and donepezil drug-administered mice. The result showed that some NTs such as γ-aminobutyric acid, acetylcholine and glutamic acid presented significant differences in the same biological matrix with different doses of the combination of GT and donepezil. In addition, based on the theory of “brain-gut-microbiome” axis, the combination of GT and donepezil with one time of clinical equivalent dose exhibited a synergistic anti-dementia effect by regulating the disordered NTs to normal levels in different biological matrices. In summary, the presented HILIC-UHPLC-QTRAP®/MS2 method might be used to monitor NTs from different clinical biological fluids in the further study.

Analysis and evaluation of nucleosides, nucleobases, and amino acids in safflower from different regions based on ultra high performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry.

Safflower has both medicinal and edible values but research on its nutrient composition is still lacking. This study was established for the quantitative determination of 28 nucleosides, nucleobases, and amino acids based on the ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry. Analysis of 30 batches of safflower from different producing areas indicated that the contents of l-proline, l-asparagine, l(+)-arginine, l-serine, l-histidine, uracil, guanosine, and uridine was high in safflower. Principle component analysis and cluster analysis found that samples from different regions could be distinguished well, and samples from the same area could be clustered into one class, different geographical environments may cause the differences of nucleosides, nucleobases, and amino acids in safflower. The analysis of principal component analysis, cluster analysis, and counter propagation artificial neural network show similar results. Then the content of nucleosides, nucleobases, and essential amino acids were compared, and found that the content in safflower from Gansu was higher than those from other regions, and there was a little difference between the samples from Xinjiang, Sichuan, and Yunnan. This research revealed the composition of nucleosides, nucleobases, and amino acids in safflower, and provided a theoretical basis for utilization of safflower.

Identification and quantification of phospholipids in strawberry seeds and pulp (Fragaria × ananassa cv San Andreas) by liquid chromatography with electrospray ionization and tandem mass spectrometry.

An extensive characterization and quantification of intact phospholipids (PLs) in strawberry (Fragaria × ananassa cv San Andreas) seed and pulp was carried out by hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization (ESI) coupled to either Fourier-transform (FT) orbital-trap or linear ion-trap tandem mass spectrometry (LIT-MS/MS). More than 150 intact polar lipids including phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), phosphatidylglycerols (PGs), phosphatidic acids (PAs), phosphatidylinositols (PIs), lysophosphatidylcholines (LPCs), and lysophosphatidylethanolamines (LPEs) were identified in negative ESI mode. PC 18:2/18:2 and 18:2/18:3 were found to be the major components of strawberry lipid extracts at concentrations of 230 ± 36 and 189 ± 32 μg/g, respectively, in seeds and at concentrations of 330 ± 50 and 140 ± 22 μg/g, respectively, in pulp. The lipidic extracts of both strawberry seeds and pulp exhibited the dominance of LPC 16:0/0:0 at a content of 132 ± 19 and 114 ± 16 μg/g, respectively, and LPC 0:0/18:2 at 236 ± 20 and 150 ± 20 μg/g, respectively. The other most abundant species of strawberry seeds and pulp were PE 18:2/18:2, 40 ± 9 and 190 ± 40 μg/g, followed by PI 16:0/18:2, 51 ± 15 and 24 ± 8 μg/g, respectively, while PG, PA, and LPE show comparable abundance below 10 μg/g. The most recurrent fatty acyl substituents of PLs were C18:3 (α-linolenic acid), C18:2 (linoleic acid), C18:1 (oleic acid), C18:0 (stearic acid), C16:0 (palmitic acid), and relatively high contents of a shorter chain such as C14:0 (myristic acid).

Hydrophilic interaction chromatography combined with ultrasound-assisted ionic liquid dispersive liquid–liquid microextraction for determination of underivatized neurotransmitters in dementia patients’ urine samples

A sensitive, rapid, precise and specific analytical method of hydrophilic interaction ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (HILIC-UHPLC–QTRAP®/MS2) combined with a high-efficiency and easy sample preparation technology of ultrasound-assisted ionic liquid dispersive liquid–liquid microextraction (UA-IL-DLLME) was developed to investigate neurotransmitters (NTs) in mild cognitive impairment, mild dementia and moderate dementia patients’ urine samples. Firstly, the UA-IL-DLLME parameters were optimized using Plackett–Burman screening and rotatable central composite design, and the main optimal conditions were obtained: ultrasound power of 307 W, ultrasound time of 4.3 min and agitation time of 4.8 min. Secondly, HILIC-UHPLC–QTRAP®/MS2 method was developed to simultaneously determine 15 underivatized NTs in urine samples. The analysis results of clinical samples showed that some NTs such as γ-aminobutyric acid (GABA), acetylcholine (Ach) and glutamic acid (Glu) presented significant differences in different dementia stages. Finally, multivariate analysis based on the combination of principal component analysis and supervised counter propagation artificial neural network was developed for comprehensive analysis of the obtained clinical data sets. As a result, GABA and Glu were simultaneously presented meaningful contribution for classification of samples, and might be considered as potential differential compounds to the urine samples from cluster patients with different dementia stages. In summary, the presented strategy of preparation, analysis and statistics might be used to investigate NTs in different clinical biological fluids.

Analysis of Phospholipids, Lysophospholipids, and Their Linked Fatty Acyl Chains in Yellow Lupin Seeds (Lupinus luteus L.) by Liquid Chromatography and Tandem Mass Spectrometry.

Hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization (ESI) coupled to either Fourier-transform (FT) orbital-trap or linear ion-trap tandem mass spectrometry (LIT-MS/MS) was used to characterize the phospholipidome of yellow lupin (Lupinus luteus) seeds. Phosphatidylcholines (PC) were the most abundant species (41 ± 6%), which were followed by lyso-forms LPC (30 ± 11%), phosphatidylethanolamines (PE, 13 ± 4%), phosphatidylglycerols (PG, 5.1 ± 1.7%), phosphatidic acids (PA, 4.9 ± 1.8%), phosphatidylinositols (PI, 4.7 ± 1.1%), and LPE (1.2 ± 0.5%). The occurrence of both isomeric forms of several LPC and LPE was inferred by a well-defined fragmentation pattern observed in negative ion mode. An unprecedented characterization of more than 200 polar lipids including 52 PC, 42 PE, 42 PA, 35 PG, 16 LPC, 13 LPE, and 10 PI, is reported. The most abundant fatty acids (FA) as esterified acyl chains in PL were 18:1 (oleic), 18:2 (linoleic), 16:0 (palmitic), and 18:3 (linolenic) with relatively high contents of long fatty acyl chains such as 22:0 (behenic), 24:0 (lignoceric), 20:1 (gondoic), and 22:1 (erucic). Their occurrence was confirmed by reversed-phase (RP) LC-ESI-FTMS analysis of a chemically hydrolyzed sample extract in acid conditions at 100 °C for 45 min.

An integrated strategy for rapid discovery and prediction of nucleobases, nucleosides and amino acids as quality markers in different flowering stages of Flos Chrysanthemi using UPLC–MS/MS and FT-NIR coupled with multivariate statistical analysis

Nucleobases, nucleosides, nucleotides and amino acids are important nutritional compounds, and some of them are positively correlated with the umami taste in scented tea. Taking these nutrients of Flos Chrysanthemi from three different flowering stages as the research object, an integrated strategy of ultra high-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UPLC–MS/MS), Fourier transform near infrared spectroscopy (FT-NIR) and multivariate statistical analysis was proposed to set up a systematic quality evaluation system for Flos Chrysanthemi from discovery to the rapid prediction of quality markers. First, a rapid, sensitive and accurate UPLC–MS/MS method was established to simultaneously determine 40 nucleobases, nucleosides, nucleotides and amino acids in three different flowering stages of Flos Chrysanthemi. The results indicated that Flos Chrysanthemi from three different flowering stages are rich in 40 nutritional compounds detected above and could be developed as functional products possessed nutraceutical and bioactive properties. Next, ten potential quality markers for three flowering stages of Flos Chrysanthemi were screened using partial least squares discriminant analysis (PLS-DA) and the plot of variable importance for projection (VIP), and the related metabolic pathways were identified for the first time. Furthermore, three of the ten potential quality markers, namely, glutamate, asparagine and aspartate were selected as key quality markers. Finally, three best PLS regression models constructed with FT-NIR were developed to predict the contents of the three key quality markers using the optimal spectral preprocessing methods and spectral interval combinations screened by synergy interval-PLS (Si-PLS), which yielded excellent prediction performance: 0.9766 ≤ R2p (coefficient of determination for prediction) ≤ 0.9858 and 6.3059 ≤ RPD (ratio of prediction to deviation) ≤ 7.3613. To sum up, the integrated quality marker discovery and FT-NIR quantitation strategy was suitable for fast quality evaluation of Flos Chrysanthemi.

The Comprehensive Evaluation of Safflowers in Different Producing Areas by Combined Analysis of Color, Chemical Compounds, and Biological Activity.

In the present study, a new strategy including the combination of external appearance, chemical detection, and biological analysis was proposed for the comprehensive evaluation of safflowers in different producing areas. Firstly, 40 batches of safflower samples were classified into class I and II based on color measurements and K-means clustering analysis. Secondly, a rapid and sensitive analytical method was developed for simultaneous quantification of 16 chromaticity-related characteristic components (including characteristic components hydroxysafflor yellow A, anhydrosafflor yellow B, safflomin C, and another 13 flavonoid glycosides) in safflowers by ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UPLC-QTRAP®/MS2). The results of the quantification indicate that hydroxysafflor yellow A, anhydrosafflor yellow B, kaempferol, quercetin, and safflomin C had significant differences between the two types of safflower, and class I of safflower had a higher content of hydroxysafflor yellow A, anhydrosafflor yellow B, and safflomin C as the main anti-thrombotic components in safflower. Thirdly, chemometrics methods were employed to illustrate the relationship in multivariate data of color measurements and chromaticity-related characteristic components. As a result, kaempferol-3-O-rutinoside and 6-hydroxykaempferol-3-O-β-d-glucoside were strongly associated with the color indicators. Finally, anti-thrombotic analysis was used to evaluate activity and verify the suitability of the classification basis of safflower based on the color measurements. It was shown that brighter, redder, yellower, more orange–yellow, and more vivid safflowers divided into class I had a higher content of characteristic components and better anti-thrombotic activity. In summary, the presented strategy has potential for quality evaluation of other flower medicinal materials.

Comparative analysis of the main active constituents from different parts of Leonurus japonicus Houtt. and from different regions in China by ultra-high performance liquid chromatography with triple quadrupole tandem mass spectrometry

A rapid, sensitive and convenient analytical method of ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UPLC-QTRAP®/MS2) was proposed for the simultaneous determination of characteristic alkaline and acidic components covering many structure types including alkaloids, phenolic acids, phenylpropanoids and flavonoids in Leonurus japonicus Houtt. (LJ). The proposed method was first reported and validated by assessing the matrix effects, linearity, limit of detections, limit of quantifications, precision, repeatability, stability and recovery of target components. The developed UPLC-QTRAP®/MS2 was successfully applied to simultaneously determine all target compounds in 38 batches of LJ from 11 different producing regions in China and five organs (including root, caulis, branch, flower and leaf) of LJ from the same stand planting base in Jiangsu Province (China). The result showed that LJ in different regions with different geographical position would affect the accumulation of different compounds, and the significant discrepancies of some target compounds were also observed in different organs of LJ due to different biosynthetic pathway and enzymes in different organs. Furthermore, both hierarchical clustering analysis and principal components analysis were performed to classify the 38 batches of LJ samples from different producing regions on the basis of target compounds. As a result, the samples could be mainly clustered into different groups, which were similar with areas classification. Overall, the presented method would be helpful for the comprehensive utilization and development of LJ resources.

The UPLC–ESI–QqQLIT–MS/MS method for quantitative determination of phytochemicals in ethanolic extracts of different parts of eight Ficus species: Development and validation

ABSTRACTFicus and validation of the ultra performance liquid chromatography–electrospray ionization hybrid triple quadrupole–linear ion trap–tandem mass spectrometry (UPLC–ESI–QqQLIT–MS/MS) method in a multiple-reaction monitoring (MRM) mode for the quantitative determination of 19 phytochemicals. The chromatographic separation of targeted phytochemicals was performed using the Waters ACQUITY UPLC BEH™ C18 column (1.7 μm, 2.1 mm × 50 mm) with 0.1% formic acid with water and acetonitrile as a mobile phase at a flow rate of 0.25 mL/min. The validation parameters showed the overall recoveries from 95.78−101.44% (RSD ≤ 3.25%), precision (intra-day: RSD ≤ 2.96%; inter-day: RSD ≤ 2.89%), linearity (R2 ≥ 0.9982), limit of detection (8.60 × 10–10−2.18 × 10–6 mg/mL), and the limit of quantitation (2.60 × 10–9–6.63 × 10–6 mg/mL) in the concentration range from 0.5 to 1000 × 10–6 mg/mL. This method was successfully applied in ethanolic extracts of different parts (fruits, leaves, and barks) of selected eight Ficus species. Quinic acid was predominant followed by rutin and chlorogenic acid among the studied nineteen phytochemicals. Ficus benjamina showed the maximum total content in fruits and leaves. The UPLC–ESI–QqQLIT–MS/MS method combined with principal component analysis (PCA) was successfully used for Ficus species discrimination on the basis of the contents of 15 compounds. The UPLC–ESI–QqQLIT–MS/MS method combined with PCA could be used for quality control.

Identification of Glycoside Compounds from Tobacco by High Performance Liquid Chromatography/Electrospray Ionization Linear Ion-Trap Tandem Mass Spectrometry Coupled with Electrospray Ionization Orbitrap Mass Spectrometry

In order to comprehensively screen and identify the glycoside compounds in tobacco, a simple, rapid and sensitive method of high performance liquid chromatography/electrospray ionization linear ion-trap tandem mass spectrometry (HPLC-ESI-LIT/MSn) coupled with electrospray ionization orbitrap mass spectrometry (ESI-Orbitrap-MS) was developed for the first time. As a result, twenty-two glycoside compounds, including eleven alcoholic glycosides, eight phenolic glycosides, two ester glycosides and an indole glycoside, were reliably identified from tobacco with high mass accuracy (within 5 mDa). Among them, four compounds were confirmed as novel molecules and other four compounds, as far as we know, were not reported previously in tobacco. This study provided a useful tool to identify the new structures of glycoside compounds in natural products, especially when there were no reference compounds available.

Chemometrics for comprehensive analysis of nucleobases, nucleosides, and nucleotides in Siraitiae Fructus by hydrophilic interaction ultra high performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry.

A rapid and sensitive hydrophilic interaction ultra high performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry method was validated for the simultaneous determination of 20 nucleobases, nucleosides, and nucleotides (within 3.5 min), and then was employed to test the functional food of Luo-Han-Guo samples. The analysis showed that the Luo-Han-Guo was rich in guanosine and uridine, but contained trace levels of the other target compounds. Chemometrics methods were employed to identify 40 batches of Luo-Han-Guo samples from different cultivated forms, regions and varieties. Unsupervised hierarchical cluster analysis and principal component analysis were used to classify Luo-Han-Guo samples based on the level of the 20 target compounds, and the supervised learning method of counter propagation artificial neural network was utilized to further separate clusters and validate the established model. As a result, the samples could be clustered into three primary groups, in which correlation with cultivated varieties was observed. The present strategy could be applied to the investigation of other edible plants containing nucleobases, nucleosides, or nucleotides.

Determination of four paraben-type preservatives and three benzophenone-type ultraviolet light filters in seafoods by LC-QqLIT-MS/MS

For the first time, an efficient and sensitive analytical method based on liquid chromatography-quadrupole linear ion trap-tandem mass spectrometry (LC-QqLIT-MS/MS) was developed for the simultaneous determination of four paraben-type preservatives and three benzophenone-type ultraviolet light filters in both plant (Sargassum fusiforme, porphyra, kelp) and animal (hairtail, yellow croaker, shrimp) seafood. The samples were extracted in methanol by pressurized liquid extraction (PLE), and the extracts were then cleaned up by mixed-mode cationic exchange (MCX) solid-phase extraction cartridges. Both isotope-labeled internal standards and matrix-matched calibration standards were used to alleviate and correct for the matrix effects, and the limits of quantification (LOQs) were 10.0μg kg−1 for all target compounds. The average recoveries were in the range of 80.6–107.8% at three spiked concentration levels (10, 50 and 100μgkg−1) with relative standard deviations (RSDs) below 8.5%. The results suggest that very limited contamination of these seven emerging contaminants, mainly associated with PCPs, occurred in these common seafoods.