Performance Liquid Chromatography-triple Quadrupole Mass Research Articles

Effect of Fermentation Duration on the Chemical Compounds of Coffea arabica from Ultra Performance Liquid Chromatography–Triple Quadrupole Mass Spectrometry and Gas Chromatography–Mass Spectrometry Analysis During the Washed Processing

Effect of Fermentation Duration on the Chemical Compounds of Coffea arabica from Ultra Performance Liquid Chromatography–Triple Quadrupole Mass Spectrometry and Gas Chromatography–Mass Spectrometry Analysis During the Washed Processing

Pharmaceuticals and personal care products contamination in the rivers of Chennai city during the COVID-19 pandemic.

Pharmaceuticals and personal care products (PPCPs) monitoring in surface water is crucial to address the escalating threat of antimicrobial resistance and safeguard public health. This study aimed to investigate the occurrence of 21 different PPCPs, including wastewater chemical markers, antibiotics, and parabens in the surface water of Chennai city using Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry (UPLC-MS/MS) analysis. It is noteworthy that chemical markers viz., carbamazepine (CBZ) and caffeine (CAF) were detected in all the sites and contributed to more than 95% of the total PPCPs load indicating a significant intrusion of wastewater. Among the antibiotics, fluoroquinolones were dominant in this study. Interestingly, a significant and strong correlation was seen between fluoroquinolones, CBZ and CAF (R2 = 0.880-0.928, p < 0.05), suggesting similarities in their sources. More than 50% of the sites exhibited a risk for antimicrobial resistance (RQAMR) withRQAMR > 1 for ciprofloxacin, indicating a significantpublic health concern. The ecotoxicological risk assessment of PPCPs showed no risk to any organisms, except for triclosan, which posed a risk to fish and daphnids at one sitenear an open drain in Buckingham canal.

An economically viable stable isotope-enhanced multiple reaction monitoring method for total fatty acid analysis in a mouse model of non-alcoholic fatty liver disease

The complex pathological mechanisms of non-alcoholic fatty liver disease (NAFLD) are closely related to dysregulated lipid metabolism, and the therapeutic effects of the traditional Chinese medicine Zexie-Baizhu Decoction (AA) on NAFLD have been gaining increasing attention. However, research into altered lipid metabolism, especially fatty acids, in NAFLD and the intervention of AA faces technical challenges, especially in the precise quantitative analysis of fatty acids in biological samples. The high complexity of biological matrices, particularly after drug intervention, greatly increases the difficulty of detection. Therefore, this study innovatively developed a simple and economical stable isotope derivatization technique by synthesizing d6N,N-dimethylethylenediamine (d6-DMED) in the laboratory, establishing a simple and economical method for fatty acid quantification. This method employs a chemical reaction under low-temperature conditions to ensure the efficient synthesis of d6-DMED. Using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry technique (UHPLC-MS/MS), combined with optimized chromatographic separation conditions and dynamic multiple reaction monitoring mode, the study established a highly sensitive detection method for 35 fatty acid derivatives. Methodological evaluation showed that the limits of quantification ranged from 0.002 to 0.060 μM, with high linearity of R² > 0.995. Additionally, the relative recovery rates were between 93.14% and 106.63%. To further demonstrate the feasibility of this method for fatty acid quantification, it was applied to measure fatty acids in multiple tissues in a mouse NAFLD model, as well as the effects of AA intervention on fatty acid metabolism. This rapid, simple, and cost-effective detection method not only enhances the understanding of NAFLD mechanisms but also provides a new strategy for evaluating the biological complex system after drug intervention.

Simultaneous determination of 21 perfluorinated and polyfluoroalkyl substances in plant oils by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry

Edible plant oils are a key component of the daily human diet, and the quality and safety of plant oils are related to human health. Perfluorinated and polyfluoroalkyl substances (PFASs) are pollutants that can contaminate plant oil through the processing of raw materials or exposure to materials containing these substances. Thus, establishing a sensitive and accurate analytical method for the determination of PFASs is critical for ensuring the safety of plant oils. In this study, a method based on acetonitrile extraction and solid phase extraction purification combined with ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS) was developed for the simultaneous determination of 21 PFASs, including perfluorocarboxylic acids, perfluoroalkyl sulfonic acids, and fluorotelomer sulfonic acids, in edible plant oils. The chromatographic conditions and MS parameters were optimized, and the influences of the extraction solvents and purification method were systematically studied. Plant oil samples were directly extracted with acetonitrile and purified using a weak anion-exchange (WAX) column. The 21 target PFASs were separated on a reversed-phase C18 chromatographic column and detected using a triple quadrupole mass spectrometer with an electrospray ionization source. The mass spectrometer was operated in negative-ion mode. The target compounds were analyzed in multiple reaction monitoring (MRM) mode and quantified using an internal standard method. The results demonstrated that the severe interference observed during the detection of PFASs in the co-extracted substances was completely eliminated after the extraction mixture was purified using a WAX column. The 21 target PFASs showed good linearity in their corresponding ranges, with correlation coefficients greater than 0.995. The limits of detection (LODs) and limits of quantification (LOQs) of the method were in the range of 0.004-0.015 and 0.015-0.050 μg/kg, respectively. The recoveries ranged from 95.6% to 115.8%, with relative standard deviations (RSDs) in the range of 0.3%-10.9% (n=9). The established method is characterized by simple sample pretreatment, good sensitivity, high immunity to interferences, and good stability, rendering it suitable for the rapid analysis and accurate determination of typical PFASs in edible plant oils.

Combined non-targeted and targeted metabolomics reveals the mechanism of delaying aging of Ginseng fibrous root.

Background: The fibrous root of ginseng (GFR) is the dried thin branch root or whisker root of Ginseng (Panax ginseng C. A. Mey). It is known for its properties such as tonifying qi, producing body fluid, and quenching thirst. Clinically, it is used to treat conditions such as cough, hemoptysis, thirst, stomach deficiency, and vomiting. While GFR and Ginseng share similar metabolites, they differ in their metabolites ratios and efficacy. Furthermore, the specific role of GFR in protecting the body remains unclear. Methods: We employed ultra-high performance liquid chromatography-triple quadrupole mass spectrometry to examine alterations in brain neurotransmitters and elucidate the impact of GFR on the central nervous system. Additionally, we analyzed the serum and brain metabolic profiles of rats using ultra-high performance liquid chromatography-quadrupole-orbitrap mass spectrometry to discern the effect and underlying mechanism of GFR in delaying aging in naturally aged rats. Results: The findings of the serum biochemical indicators indicate that the intervention of GFR can enhance cardiovascular, oxidative stress, and energy metabolism related indicators in naturally aging rats. Research on brain neurotransmitters suggests that GFR can augment physiological functions such as learning and memory, while also inhibiting central nervous system excitation to a certain degree by maintaining the equilibrium of central neurotransmitters in aged individuals. Twenty-four abnormal metabolites in serum and seventeen abnormal metabolites in brain could be used as potential biomarkers and were involved in multiple metabolic pathways. Among them, in the brain metabolic pathways, alanine, aspartate and glutamate metabolism, arginine and proline metabolism, histidine metabolism, and tyrosine metabolism were closely related to central neurotransmitters. Butanoate metabolism improves energy supply for life activities in the aging body. Cysteine and methionine metabolism contributes to the production of glutathione and taurine and played an antioxidant role. In serum, the regulation of glycerophospholipid metabolism pathway and proline metabolism demonstrated the antioxidant capacity of GFR decoction. Conclution: In summary, GFR plays a role in delaying aging by regulating central neurotransmitters, cardiovascular function, oxidative stress, energy metabolism, and other aspects of the aging body, which lays a foundation for the application of GFR.

Unambiguous identification of γ-aminobutyric acid adducts as novel plant biomarkers and their ultra-sensitive detection by UPLC–MS/MS for retrospective analyses of nitrogen mustards exposure

Plants are widely existing in the environments and have been considered as potential sentinel species of toxic chemicals′ exposure. In this study, the deadly toxic chemicals of three nitrogen mustards (NMs, including NH1, NH2 and NH3) were selected as the investigated targets. First, the reactivities of common endogenous plant components with NMs were examined in vitro. Then, the model plant Nicotiana benthamiana Domin was exposed to NMs. Three γ-aminobutyric acid–nitrogen mustard adducts (GABA–NMs) were identified in the living plant by high resolution mass spectrometry and comparison with the synthesized references. A sensitive detection method with the limits of quantification of 0.0500 ng mL−1 was developed using ultrahigh performance liquid chromatography–triple quadrupole mass spectrometry. The GABA–NMs could be detected after 120 days of the exposure and even in the dead leaves without obvious decrease. Furthermore, 20 different plant species grown in diverse climate zones were exposed to HN1, and the adduct of GABA–HN1 was identified in all the leaves. The results showed the good universality and specificity of GABA–NMs as plant biomarkers for NMs exposure. This work provides a new approach for the pollution investigation of toxic chemicals through analysing biomarkers in plant materials.

Simultaneous determination of six benzodiazepine sedatives residue in aquatic products by high performance liquid chromatography-triple quadrupole mass spectrometry

To establish a method for the simultaneous determination of 6 benzodiazepine sedatives residue in aquatic products by high performance liquid chromatography-triple quadrupole mass spectrometry. The samples were extracted with acetonitrile and purified by C_(18 )solid phase extraction column. The sample solution was separated by Waters ACQUITY UPLC BEH C_(18 )column(2.1 mm×50 mm, 1.7 μm) using 0.1% formic acid and methanol as mobile phase for gradient elution, determined in multiple reaction monitoring mode and quantified by internal standard method. Six benzodiazepine sedatives had a good linear relationship in the range of 1.0-50.0 μg/L with r&gt;0.9990, the limits of detection and limits of quantification were 0.3 and 1.0 μg/kg. Average recoveries for the analytes at 3 spiked levels ranged from 74.2%-108.0% with relative standard deviations of 1.1%-6.7%(n=6). The method is simple, rapid, sensitive and accurate, which is suitable for simultaneous determination of 6 benzodiazepine sedatives residue in aquatic products.

Determination of 145 pharmaceuticals and personal care products in eleven categories in water by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry

Pharmaceuticals and personal care products (PPCPs) are emerging contaminants frequently detected in aquatic environments at trace levels. These chemicals have diverse structures and physicochemical properties and includes pharmaceuticals like antibiotics, antihypertensive drugs, antiviral drugs, and psychotropic drugs that are widely used in large quantities worldwide. Considering the large number of pharmaceuticals currently in usage, it is crucial to establish a priority list of PPCPs that should be monitored and/or treated first. An accurate understanding of the occurrence and levels of PPCPs in aquatic environments is essential for providing objective materials for monitoring these emerging contaminants. Therefore, accurate, efficient, sensitive, and high-throughput screening techniques need to be established for determining and quantifying PPCPs. This study developed a method for the determination of 145 PPCPs (grouped into eleven categories: antibiotics, antihypertensive drugs, antidiabetic drugs, antiviral drugs, β-receptor agonists, nitroimidazoles, H2 receptor antagonists, psychotropic drugs, hypolipidemic drugs, non-steroidal anti-inflammatory drugs, and others) in water. The method was based on large volume direct injection without sample enrichment and cleanup and used ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS). Water samples were collected and filtered through a 0.22-μm regenerated cellulose (RC) filter membrane. Subsequently, Na2EDTA was added to the samples to adjust their pH to 6.0-8.0. Internal standards were mixed with the solutions, and because of the addition of Na2EDTA, the interference of metal ions could be eliminated in the determination of compounds, especially for tetracycline and quinolone antibiotics. Among the six filter membranes tested in this study (PES, PFTE-Q, PFTE, MCE, GHP, and RC), RC filter membranes were screened for water sample filtration. The UHPLC-MS/MS parameters were optimized by comparing the results of various mobile phases, as well as by establishing the best instrumental conditions. The 145 PPCPs were separated using an Phenomenex Kinetex C18 column (50 mm×3 mm, 2.6 μm) via gradient elution. The mobile phases were 0.1% (v/v) formic acid aqueous solution containing 5 mmol/L ammonium formate and acetonitrile for positive ion modes, 5 mmol/L aqueous solutions of ammonium formate and acetonitrile for negative ion modes. The samples were quantified using the scheduled multiple reaction monitoring (scheduled-MRM) mode with electrospray ionization in positive and negative ion modes. A standard internal calibration procedure was used to calculate contents of sample. The established method was systematically verified, and it demonstrated a good linear relationship. The average recoveries of the 145 PPCPs at the three spiked levels were in the range of 80.4%-128% with relative standard deviations (RSDs, n=6) of 0.6%-15.6%. The method detection limits (MDLs) ranged from 0.015 to 5.515 ng/L. Finally, the optimization method was applied to analyze the 145 PPCPs in 11 surface water samples and 6 drinking water samples. Overall, 93 (64%) out of the 145 analytes were detected. The total contents of the PPCPs in surface water samples ranged from 276.9 to 2705.7 ng/L. The detection frequencies of antidiabetic, antiviral, and psychotropic drugs were 100%. The total contents of the PPCPs in drinking water samples ranged from 140.5 to 211.5 ng/L, and antibiotics, antidiabetic drugs, and antiviral drugs comprised the largest proportion of analytes (by mass concentration) in drinking water samples. Our method exhibited high analytical speed and high sensitivity. It is thus suitable for the trace analysis and determination of the 145 PPCPs in environmental water and showed improved detection efficiency for PPCPs in water, indicating that it has a high potential for practical applications. This study can extend technical support for further pollution-level analysis of PPCPs in water and provide an objective basis for environmental management.

Rapid electrochemical detection of L-lactate in Baijiu affecting serotonin and dopamine secretion in mice.

Baijiu, a traditional Chinese alcoholic beverage, carries China's rich historical and cultural heritage. Consumers experience varying levels of relaxation and pleasure after consuming different types of Baijiu, with the biological basis of delectation influenced by serotonin and dopamine. In this study, we prepared carbon fiber electrodes modified with surface decorated gold nanoparticles to directly measure the electrochemical response signals in the serum of mice before and after gavage with different types of Baijiu. It was observed that the serum signal change in mice after consuming Baijiu sample 1 (J1) was higher than that of the other two types of Baijiu. Consequently, trace flavor compounds in the Baijiu samples were detected using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS), revealing the highest content of L-lactic acid in J1. Mice were intraperitoneally injected with 200 mg kg-1 of L-lactic acid. The changes in dopamine and serotonin in the serum of the injected mice were monitored using a biosensor, and the results were compared with the results of high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS). The findings confirmed that L-lactic acid could indeed stimulate the secretion of both neurotransmitters in mice, suggesting that the trace components in J1 may even exhibit synergistic effects. This study contributes to a deeper understanding of the effects of Baijiu on the body and provides a scientific basis for the production and consumption of Baijiu.

Determination of 118 pesticide residues in urine by ultra-high performance liquid chromatography-tandem mass spectrometry

Pesticide residues in food and their hazardous effects have attracted much attention given the increased and widespread use of pesticides. The long-term consumption of food containing pesticide residues is an important pathway for the gradual accumulation of pesticides in the human body. Urine is often monitored as a biological sample for low-dose exposure to pesticides, and urine collection is a relatively convenient sampling technique in general population research. In order to effectively monitor residual levels of multiple pesticides in human urine and provide an important technological approach for health risk assessment, a rapid screening and confirmatory detection method for 118 pesticides in urine was established using QuEChERS method as a pretreatment combined with ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS). The 118 pesticides analyzed included organophosphorus, carbamate, neonicotinoid, and strobilurin fungicides and other widely used pesticides. Following systematic optimization of the pretreatment process, LC separation conditions, and MS/MS parameters, 118 pesticides were extracted from urine samples and analyzed within 2 h. In brief, the target analytes in 5 mL urine samples were extracted with 10 mL of acetonitrile and added with 5 g of anhydrous MgSO4 and 1 g of NaCl as water-removal and salting-out agents, respectively. After centrifugation, 6 mL of the supernatant was cleaned using the QuEChERS method with 300 mg of C18, 300 mg of primary secondary amine (PSA) and 900 mg of anhydrous MgSO4 as the purification adsorbent. After nitrogen blowing and solubilization, the 118 target analytes were separated on a ZORBAX Eclipse Plus C18 analytical chromatographic column (100 mm×2.1 mm, 1.8 μm) with gradient elution using (A) 0.01% formic acid aqueous solution (containing 2 mmol/L ammonium formate) and (B) 0.01% formic acid methanol solution (containing 2 mmol/L ammonium formate) as mobile phases. The gradient elution program was as follows: 0-0.5 min, 5%B; 0.5-1.5 min, 5%B-20%B; 1.5-2.5 min, 20%B-50%B; 2.5-8.0 min, 50%B-80%B; 8.0-9.0 min, 80%B-98%B; 9.0-11.0 min, 98%B; 11.0-11.5 min, 98%B-5%B; 11.5-15.0 min, 5%B. The analytes were then determined by UHPLC-MS/MS with positive/negative ion switching in dynamic multiple-reaction monitoring mode and quantified using the external standard method. The results indicated that the proposed method can determine 118 pesticides in urine simultaneously and rapidly. The limits of detection and limits of quantification were 0.10 and 0.50 μg/L, respectively, and the matrix effects were less than 20%for all targeted compounds. The recoveries of the 118 pesticides in urine were between 70.2% and 104% at three spiked levels of 0.50, 1.00, and 5.00 μg/L, and the relative standard deviations ranged from 2.8% to 9.3%. The method was applied to 10 actual urine samples, and the results revealed the presence of six pesticides, including thiamethoxam, clothianidin, acetamiprid, dinotefuran, isoproturon, and dimethomorph, with contents ranging from <LOQ to 3.65 μg/L. Thiamethoxam and clothianidin were detected most often in the samples, and their concentrations were highly correlated with the current use of these pesticides in edible agricultural products. The proposed method has the characteristics of high efficiency, sensitivity, and accuracy and is suitable for the rapid screening and determination of the 118 pesticide residues in human urine samples.

Characteristics and Ecological Risk Assessment of Pharmaceutical Active Compounds in Guangdong Province

Pharmaceutically active compounds(PhACs) have become a class of new pollutants in the environment after extensive production and use of PhACs in China. To investigate the pollution characteristics of PhACs in Guangdong Province, raw sewage was collected from 186 sewage treatment plants in 21 cities, including 178 townships and administrative districts in Guangdong Province. The pollution levels of ten typical PhACs in influent water of sewage treatment plants were analyzed using automatic solid phase extraction and high performance liquid chromatography-triple quadrupole mass spectrometry. The spatial distribution characteristics of PhACs in Guangdong Province were fully revealed, and the potential ecological risks of PhACs were evaluated. The results showed that PhACs were detected in all wastewater plants, and the mass concentration of PhACs ranged from 21.00 to 9558.25 ng·L-1. Metoprolo, acetaminophen, bezafibrate, and caffeine were the main pollutants. In terms of spatial distribution, the average mass concentration of ΣPhACs in various regions of Guangdong Province was in the following order:Pearl River Delta>North Guangdong>East Guangdong≈West Guangdong. When the mass concentration of ΣPhACs was over 2500 ng·L-1 in the influent water of sewage treatment plants, the concentration of PhACs in effluent was estimated according to the sewage disposal technology. The ecological risk of PhACs was carried out based on the effluent. The results revealed that the ecological risk of PhACs was low in Guangdong Province, and the risk of bezafibrate was moderate in the cities of Shaoguan, Jiangmen, and Shenzhen. The highest ecological risk of ΣPhACs was located in Shaoguan.

Quality evaluation of Angelica Sinensis Radix dispensing granules by integrating microvascular activity and chemical analysis

Ethnopharmacological relevanceAs a new form of crude slices of traditional Chinese medicine (TCM), traditional Chinese medicine dispensing granules (TCMDGs) have been used for clinical formula. It is necessary to evaluate whether the chemical composition and biological activity are consistent among the different batches. Angelica Sinensis Radix (ASR), the root of Angelica sinensis (Oliv.) Diels, is one of the most frequently used medicinal materials in gynecology, senile and cardiovascular diseases. In this paper, the quality of TCMDGs is examined taking the Angelica Sinensis Radix dispensing granules (ASRDGs) as a typical case. Aim of the studyIn this study, integrating bioequivalence and chemical analysis was used to evaluate the quality of dispensing granules taking ASRDGs as a typical case. Materials and methodsAccording to the clinical efficacy of ASR, the intestinal absorption liquid of ASRDGs (IAL-ASRDGs) in 15 batches prepared by the everted gut sac (EGS) method was used to evaluate its effects of vasodilatation on isolated vascular rings. Then, the chemical profiling analysis of IAL-ASRDGs from the 15 batches was carried out by ultra-high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS). The components in IAL-ASRDGs were identified using mass spectrometry data and the obtained reference standards. Pearson correlation analysis was further performed for the selection of quality control markers based on the extracted ion chromatograms of the identified compounds and vasodilatory activities of different batches of IAL-ASRDGs. Moreover, the selected chemical components in ASRDGs were further verified by vasodilatory activities and quantitatively analyzed by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQ-MS). ResultsThe IAL-ASRDGs showed favorable vasodilatory activities. There were significant differences among the 15 batches. The ranges of maximum vasodilation rate (%) and EC50 were 69.33 ± 7.16 to 19.52 ± 5.05 and 0.07–25.58 g raw materials/mL, respectively. A total of 46 compounds in IAL-ASRDGs were identified based on accurate mass measurements, fragmentation behavior and the reference standards. Among them, 8 compounds including butylidenephthalide, butylphthalide and senkyunolide A showed positive correlation with vasodilatory effect of IAL-ASRDGs. The 8 compounds were further verified, showing obvious vasodilatory activities. The content of the 8 compounds also showed some differences in 15 batches of ASRDGs. Among them, the content of ferulic acid, senkyunolide I and senkyunolide H varied the most in different batches of ASRDGs. By Pearson correlation analysis, the total content of senkyunolides (senkyunolide A, senkyunolide I and senkyunolide H) in ASRDGs was found to reflect the vasodilatory activity of ASRDGs mostly. ConclusionThis study provides new strategies for the quality assessment by bioequivalence and explore the chemical quality control markers for ASRDGs.

Determination of 13 halobenzoquinone disinfection by-products in drinking water using solid phase extraction-ultra performance liquid chromatography-triple quadrupole mass spectrometry

Disinfection of drinking water is critical to prevent waterborne diseases. An unexpected consequence of water disinfection is the formation of disinfection by-products by the interaction of disinfectants with organic matter (natural or anthropogenic) and halides, which present significant toxicological effects and carcinogenic risks. As an emerging disinfection by-product, halobenzoquinones (HBQs) have attracted increasing attention owing to their severe toxicity and high detection rates. The credible determination of HBQs is essential for further studies on their occurrence, toxicity, and control measures; however, HBQs are usually detected in drinking water at trace levels. Therefore, accurate and efficient analytical techniques are critical for HBQ determination and quantitation. In this study, a method based on solid phase extraction (SPE) combined with ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS) was developed to determine 13 HBQs, including six chlorobenzoquinones, six bromobenzoquinones, and one iodobenzoquinone, in drinking water. One-liter water samples were added with 2.5 mL of formic acid, and 500 mL of each sample was collected for further enrichment. Pretreatment optimization mainly focused on the SPE column, washing solvent, and nitrogen blowing temperature. After extraction using Plexa SPE columns (200 mg/6 mL), the samples were washed with ultrapure water containing 0.25% formic acid combined with 30% methanol aqueous solution containing 0.25% formic acid, eluted with 6 mL of methanol containing 0.25% formic acid, and then nitrogen blown at 30 ℃. The UPLC-MS/MS parameters were optimized by comparing the results of two reversed-phase columns (BEH C18 and HSS T3) and various concentrations of formic acid in the mobile phase, as well as by establishing the best instrumental conditions. The separation of 13 HBQs was performed using an HSS T3 column (100 mm×2.1 mm, 1.8 μm) via gradient elution with a mixture of 0.1% formic acid aqueous solution and methanol as the mobile phase for 16 min. The 13 HBQs were detected using a triple quadrupole mass spectrometer equipped with a negative electrospray ionization source (ESI-) in multiple reaction monitoring (MRM) mode. Matrix-matched calibration curves were used to quantify the HBQs owing to intense matrix inhibitory effects. The results reflected the good linear relationships of the 13 HBQs and yielded correlation coefficients (r) greater than 0.999. The method detection limits (MDLs, S/N=3) were 0.2-10.0 ng/L, while the method quantification limits (MQLs, S/N=10) were 0.6-33.0 ng/L. The recoveries of the 13 HBQs were 56%-88% at three spiked levels (10, 20, 50 ng/L), and the relative standard deviations (RSDs, n=6) were less than or equal to 9.2%. The optimization method was applied to analyze HBQs in five drinking water samples. Four HBQs, namely, 2,6-dichloro-1,4-benzoquinone (2,6-DCBQ), 2,5-dibromo-1,4-benzoquinone (2,5-DBBQ), 2,6-dibromo-1,4-benzoquinone (2,6-DBBQ), and 2,6-dibromo-3,5-dimethyl-1,4-benzoquinone (2,6-DBDMBQ), were detected in the samples with detection rates of 100%, 20%, 80%, and 20%, respectively. The most frequently detected HBQ, 2,6-DCBQ, also exhibited the highest content (15.0-56.2 ng/L). The method showed high sensitivity, stability, accuracy, and efficiency, rendering it suitable for the analysis of 13 HBQs in drinking water. Compared with previous methods that mainly focused on 2,6-DCBQ and 2,6-DBBQ, the developed method achieved higher throughput and enabled the simultaneous analysis of 13 HBQs. The method presented in this study provides an opportunity to explore different types and concentrations of HBQs in drinking water, offers a deeper understanding of the occurrence of HBQs, and facilitates further studies on the health risks and control measures of these compounds.

Effect of BBM gene on callus growth and ginsenoside content in Panax quinquefolius

Baby Boom(BBM) gene is a key regulatory factor in embryonic development and regeneration, cell proliferation, callus growth, and differentiation promotion. Since the genetic transformation system of Panax quinquefolius is unstable with low efficiency and long period, this study attempted to transfer BBM gene of Zea mays to P. quinquefolius callus by gene gunship to investigate its effect on the callus growth and ginsenoside content, laying a foundation for establishing efficient genetic transformation system of P. quinquefolius. Four transgenic callus of P. quinquefolius with different transformation events were obtained by screening for glufosinate ammonium resistance and molecular identification by PCR. The growth state and growth rate of wild-type and transgenic callus were compared in the same growth period. The content of ginsenoside in transgenic callus was determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS). The results showed that transgenic callus growth rate was significantly higher than that of wild-type callus. In addition, the content of ginsenoside Rb_1, Rg_1, Ro, and Re was significantly higher than that in wild-type callus. The paper preliminarily proved the function of BBM gene in promoting growth rate and increasing ginsenoside content, which provided a scientific basis to establish a stable and efficient genetic transformation system for Panax plants in the future.

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

Ethnopharmacological relevanceJiuwei 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 studyThis 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 methodsIn 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). ResultsA 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. ConclusionsJWXF 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.

Identification of Nutritional Ingredients and Medicinal Components of Hawk Tea and Insect Tea Using Widely Targeted Secondary Metabolomics

In this study, the metabolites in insect tea and hawk tea were analyzed by ultra-high performance liquid chromatography–triple quadrupole mass spectrometry. We found 49 metabolites in insect tea and hawk tea that can be used as key active components in traditional Chinese medicine, as well as 98 metabolites that can be used as active components of pharmaceutical preparations for the treatment of cancers, hypertension, cardiovascular diseases, etc. Comparative analysis found that insect tea and hawk tea had significant differences in their metabolic profiles. Insect tea contains more metabolites beneficial to human health than hawk tea; insect tea also has higher antioxidant activity in vitro than hawk tea. The results of this study will contribute to the development of new health foods using insect tea.

Pseudotargeted metabolomics analysis of pine pollen intervention in the liver of premature ovarian failure rats

Premature ovarian failure (POF) is a prevalent gynecological disease. In traditional Chinese medicine, it is believed that POF is directly related to abnormal function of the liver and kidneys. As such, regulation of the liver metabolism through the use of medicinal and edible substances is important for the treatment of POF. Pine pollen, a traditional Chinese medicinal and edible pollen variety, contains various active substances, such as sex hormones and phytohormones, which have been used to inhibit inflammation, regulate the immune system, and protect reproductive tissues. Using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS), this study examined the influence of pine pollen on the liver metabolome of cyclophosphamide-induced POF model Sprague Dawley (SD) rats. The variations in the metabolites present in the liver tissue of control SD rats, model SD rats, and SD rats treated with various doses of pine pollen or estrogen were analyzed using principal component analysis (PCA) in combination with orthogonal partial least squares discriminant analysis (OPLS-DA) and other multivariate statistical methods to reveal the mechanism of pine pollen intervention in the livers of POF SD rats. An animal model experiment was conducted using six groups of ten-week-old rats. Cyclophosphamide was administered intraperitoneally to the model group and four intervention groups at a dosage of 60 mg/kg for 1 d followed by a dosage of 10 mg/kg for 14 d. Within the following four weeks, each of the four intervention groups received the intragastric administration of 0.1, 0.5, or 1.5 g/kg bodyweight (BW) of pine pollen, or 0.075 g/kg BW of conjugated estrogens (positive control). Equal quantities of normal saline were administered to the control and cyclophosphamide-treated model groups. Subsequently, the rat livers were subject to pseudotargeted metabolomics, and a total of 687 liver metabolites were discovered using both positive and negative ions. The metabolites differing in content were screened using the t-test (p<0.05) and the fold change (FC>2 or <0.5) in univariate analysis, and the variable importance in projection (VIP>1) in multivariate analysis. It was found that in comparison with the control group, the contents of 32 metabolites significantly increased, while those of 28 metabolites significantly decreased in the model group. The majority of these metabolites were involved α-linolenic acid metabolism, vitamin B6 metabolism, and purine metabolism, along with the lysine degradation and glycolysis/gluconeogenesis metabolic pathways. Compared with the cyclophosphamide-induced model group, the estrogen group exhibited increased levels of 47 metabolites and decreased levels of 29 metabolites, wherein 34 metabolites were restored to the levels found in the control group. These metabolites mainly involved the vitamin B6, lysine, glycolysis/gluconeogenesis, arginine and proline, and cysteine and methionine metabolic pathways. In the low/medium/high-dose pine pollen groups, the contents of 34/32/34 metabolites increased, the contents of 30/37/24 metabolites decreased, and the contents of 47/38/34 metabolites were restored to the levels found in the control group, respectively. These metabolites were mainly involved in vitamin B6 metabolism, purine metabolism, and the glycolysis/gluconeogenesis metabolic pathway. These results therefore indicate that the restoring effect of pine pollen is equivalent or superior to that of conjugated estrogen. Additionally, based on the known metabolic pathways, it appears that when estrogen interferes with the liver metabolism, the key metabolic pathways that become affected are the arginine and proline metabolism and cysteine and methionine metabolism pathways. In contrast, pine pollen intervention affected existing metabolic pathways that were known to be disordered by cyclophosphamide. The use of pine pollen may therefore restore the levels of many metabolites. It should be noted that 23 overlaps exist between the estrogen-restored metabolites and the pine pollen-restored metabolites, including a variety of acylcarnitines, such as ACar 10∶0. As a result, pine pollen extract may be able to normalize the liver metabolic abnormalities induced by POF. This study therefore establishes a theoretical reference for the development of functional applications for pine pollen and for the treatment of POF.

Determination of 107 typical pesticides and metabolites in raw water and drinking water by online-solid phase extraction coupled with ultra performance liquid chromatography-triple quadrupole mass spectrometry

In order to monitor the risk of pesticide pollutants in drinking water, an analytical method based on online-solid phase extraction coupled with ultra performance liquid chromatography-triple quadrupole mass spectrometry (online-SPE-UPLC-MS/MS) was established for the simultaneous rapid screening and determination of 107 pesticides and metabolites (organophosphorus, organic nitrogen, organic heterocycle, carbamate, amide, benzoyl urea, neonicotinoid, etc.) in raw water and drinking water. Different injection volumes (5, 10, and 15 mL) were compared. The detection response increased with an increase in the injection volume, but the matrix effect also became more pronounced. Under the premise of ensuring the sensitivity of the method and meeting the detection requirements, the injection volume was selected as 5 mL. Accordingly, the samples were filtered through a 0.22-μm hydrophilic polytetrafluoroethylene filter, and then, 5 mL samples were injected into the online-SPE system by the automatic sampler. After adsorption on an X Bridge C18 online-SPE column, the samples were washed with pure water and eluted by gradient elution using acetonitrile and 0.1% formic acid aqueous solution as the mobile phases, with separation on an ACQUITY HSS T3 column. The samples were detected by multiple reaction monitoring with electrospray ionization in positive and negative ion modes, and quantified by an external standard method. Using raw water and drinking water as the sample matrices, the accuracy and precision of the method were verified. The 107 pesticides and metabolites showed good linear relationships in different ranges with correlation coefficients (r2)>0.995. The limits of detection (LODs, S/N=3) of the method were 0.03-1.5 ng/L, and the limits of quantification (LOQs, S/N=10) were 0.1-5.0 ng/L. The target pesticides were spiked at concentration levels of 1, 20, and 50 ng/L. The spiked recoveries of the 107 targets in raw water and drinking water samples were 60.6%-119.8% and 61.2%-119.0%, respectively. The corresponding relative standard deviations (RSDs, n=6) were 0.3%-18.6% and 0.4%-17.1%. The pesticide residues in raw water and drinking water were determined by this method. Amide herbicides, triazine herbicides, triazole insecticides, carbamate insecticides, and neonicotinoid insecticides had high detection rates. The detected concentrations ranged from 0.1 to 97.1 ng/L in raw water and from 0.1 to 93.6 ng/L in drinking water. The sample consumption of online-SPE method was lower than that in the traditional off-line SPE methods, which greatly improved the convenience of sample collection, storage, and transportation. The samples only need to be filtered before injection and analysis. The method is simple to operate and shows good reproducibility. With this online-SPE method, only 23 min were required from online enrichment to detection completion. The developed method has the advantages of high analytical speed and high sensitivity. The method is suitable for the trace analysis and determination of 107 typical pesticides in raw water and drinking water, which effectively improves the detection efficiency of pesticides in water and has high potential for practical application. It can extend technical support for the pollution-level analysis of typical pesticides and metabolites in drinking water and provide an objective basis for human health risk assessment.

Residue behavior and risk assessment of pyraclostrobin and tebuconazole in peppers under different growing conditions.

This study evaluates the residue behavior and risks of pyraclostrobin and tebuconazole in peppers. An analytical method for the simultaneous determination of the concentration of these fungicides in peppers was developed using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry. Pepper samples were extracted with acetonitrile and cleaned with primary secondary amine and graphitized carbon black. The average recoveries of pyraclostrobin and tebuconazole under three fortification levels were 86.7-101.4% and 81.7-104.4%, with relative standard deviations of 4.0-7.2% and 3.8-10.9%, respectively. The limit of quantification of both fungicides in peppers was 0.01mg/kg. The terminal residue trial of 30% pyraclostrobin and tebuconazole suspension concentrate was investigated for samples cultivated in open fields and greenhouses. The results showed that the terminal residues of pyraclostrobin and tebuconazole in peppers were lower than the maximum residue limits established by GB 2763-2021 (0.5mg/kg for pyraclostrobin and 2mg/kg for tebuconazole). The results of a statistical t-test indicated that there was no significant difference between samples grown in open fields and greenhouses. According to the international estimate of short-term intake (IESTI) calculation model, provided by the Joint FAO/WHO Meeting on Pesticide Residues, the acute dietary exposure risk of both fungicides in peppers was acceptable for the general population, with an IESTI of 0-3% and 0-5% of the acute reference dose for pyraclostrobin and tebuconazole, respectively.

Residue levels and risk assessment of pesticides in litchi and longan of China

A total of 57 pesticide residues were analyzed in 155 litchis and 153 longan samples collected from the three main producing provinces of China. They were studied by the QuEChERS extraction method based on analysis using ultra performance liquid chromatography–triple quadrupole mass spectrometry (UPLC–MS/MS) and gas chromatography (GC) techniques. Health risks from litchis and longans for humans were evaluated by hazard quotient (HQ) and hazard index (HI). Sixteen different pesticides were detected in litchi samples. In longan samples, 19 different pesticides were detected. A total of 72% litchi samples and 52% longan samples contained at least one pesticide residue. Carbendazim, chlorbenzuron and diflubenzuron were found in 25.2%, 28.4%, and 34.8% of the litchi samples, respectively, and 21.6%, 11.8%, and 21.6% of the longan samples, respectively. The banned pesticides carbofuran and omethoate were detected in two litchi samples and one longan sample, respectively. Residue concentrations did not exceed the maximum residue levels (MRLs) in all litchi and longan samples except for carbofuran. Dietary risk assessment results showed that the chronic and acute dietary risks (HQc and HQa) of individual pesticides and the cumulative chronic dietary risks (HIc) of all pesticide residues for adults and children were all less than 100%. However, the cumulative acute dietary risk (HIa) of all pesticide residues from litchi samples for children was 99.85%, which was close to the unacceptable level and should be given more attention. The contributions of insecticide to HIc and HIa were 89.3% and 87.3% in litchi samples and 91.7% and 55.0% in longan samples.