Potential mutagenic impurities in Active Pharmaceutical Ingredient, Meropenem Trihydrate were assessed and a novel analytical method for their quantification was developed and validated. This Liquid Chromatographic me...Potential mutagenic impurities in Active Pharmaceutical Ingredient, Meropenem Trihydrate were assessed and a novel analytical method for their quantification was developed and validated. This Liquid Chromatographic method using High Resolution Mass Spectrometer (LC-HRMS) technique is proved to be suitable for simultaneous quantification of all ten identified impurities with required specificity, sensitivity, resolution, precision, accuracy, and other method characteristics as per ICH Guidelines. The acceptable limit of less than 2.9 μg/g was considered for evaluations, based on drug substance dosage and duration of treatment. The method stands most sensitive with a Limit of Detection of 0.35 μg/g, considering the challenge full acceptance criteria as per current regulatory standards.展开更多
A simple and high-throughput method to simultaneously determine selected benzodiazepines (i.e., diazepam, lorazepam, clonazepam, and bromazepam) in urine was developed and validated. The entire methodology consisted o...A simple and high-throughput method to simultaneously determine selected benzodiazepines (i.e., diazepam, lorazepam, clonazepam, and bromazepam) in urine was developed and validated. The entire methodology consisted of the application of an innovative extraction/cleanup procedure, namely liquid-liquid extraction with low-temperature partitioning (LLE-LTP), and analysis by liquid chromatography combined with high-resolution mass spectrometry (LC-HRMS). The LLE-LTP procedure was optimized via factorial design and by evaluating crucial variables, specifically the freezing mode (either slow or fast), the urine/acetonitrile volume ratio, and the sample ionic strength. The benzodiazepines were quantified using matrix-matched calibration curves where the following parameters were assessed by validation protocol: in general, linearity range of 17 - 200 μg?L–1 (r > 0.9957);limits of detection lower than 5 μg?L–1;relative standard deviations (RSD) lower than 12.5%;and accuracy ranging from 72.3 to 117%. To test this procedure’s performance, the method was applied to determine the content of diazepam in actual urine samples. The validation results obtained for the method demonstrated that the present methodology could be potentially applied in proficient laboratories as a routine approach for determining benzodiazepines compounds content in urine.展开更多
N-Methyl bis(2-(alkyloxy-alkylphosphoryloxy)ethyl)amines, which are abbreviated as PNPs, are a series of new skeleton chemicals belonging to schedule 2.B.04 chemicals of Chemical Weapons Convention (CWC). PNPs are imp...N-Methyl bis(2-(alkyloxy-alkylphosphoryloxy)ethyl)amines, which are abbreviated as PNPs, are a series of new skeleton chemicals belonging to schedule 2.B.04 chemicals of Chemical Weapons Convention (CWC). PNPs are important markers of chemical warfare agents because they are structurally relative to both nerve agents and N-mustards. In this study, fragmentation pathways of the most characteristic fragment ions in Q-TOF mass spectrometry were proposed based on the information from accurate mass and secondary fragmentations of product ions scan experiments. Results indicated that the base ion in LC/HRMS was the quasi-molecular ion [M+H]+. In LC-HRMS/MS, it was [M+H-CnH2n+1P(O)(OH)CmH2m+1O]+ fragment ion which was formed by losing an alkyloxy alkylphosphoryloxy group from the quasi-molecular ion. The diagnostic ion m/z84.0814 was identified as [C5H10N]+, which was the group of (CH2=CH)2N+(H)CH3. PNPs have two protonated centers. One is on the N atom, the other is on the O atom (P=O). O-n-propyl PNPs generally exhibited two fragmentation pathways. Firstly, the quasi-molecular ion [M+H]+ lost a propoxy alkylphosphoryloxy group to produce [R1P(OH+)(O-n-C3H7)OCH2CH2N(CH3)CH=CH2]+, which could be fragmented further to produce [C5H10N]+ ion. Secondly, [R1P(OH+)(O-n-C3H7) OCH=CH2]+ ions were produced from [M+H]+ and fragmented further to produce the abundant ions [R1P(OH+)(OH)OCH =CH2]. However, O-isopropyl PNPs characteristically produced weak fragment ions [M+H-C3H6]+, which were presumably formed via loss of CH3CH=CH2 from [M+H]+. Other PNPs showed similar fragmentation pathways as O-n-propyl PNPs. On the summarization of the MS fragmentation pathways of PNPs, LC-HRMS/MS quantitative and qualitative methods were developed and applied to analyze N-Methyl bis(2-(butoxy-methylphosphoryloxy)ethyl]amine in high background organic samples. The analytical results had successfully supported the sample preparation for the 33rd official proficiency test of Organization for Prohibition of Chemical Weapons (OPCW).展开更多
Collagen structure in biological tissues imparts its intrinsic physical properties by the formation of several covalent crosslinks.For the first time,two major crosslinks in the skin dihydroxylysinonorleucine(HLNL)and...Collagen structure in biological tissues imparts its intrinsic physical properties by the formation of several covalent crosslinks.For the first time,two major crosslinks in the skin dihydroxylysinonorleucine(HLNL)and histidinohydroxymerodesmosine(HHMD),were isotopically labelled and then analysed by liquid-chromatography high-resolution accurate-mass mass spectrometry(LC-HRMS)and small-angle neutron scattering(SANS).The isotopic labelling followed by LC-HRMS confirmed the presence of one imino group in both HLNL and HHMD,making them more susceptible to degrade at low pH.The structural changes in collagen due to extreme changes in the pH and chrome tanning were highlighted by the SANS contrast variation between isotopic labelled and unlabelled crosslinks.This provided a better understanding of the interaction of natural crosslinks with the chromium sulphate in collagen suggesting that the development of a benign crosslinking method can help retain the intrinsic physical properties of the leather.This analytical method can also be applied to study artificial crosslinking in other collagenous tissues for biomedical applications.展开更多
目的随着新型合成大麻素的演变,其在体内代谢速度增快,对原药的检测难度增加,为确定是否吸食合成大麻素,其生物标志物可以作为强有力的证据,常见的代谢模型有体内代谢模型和体外代谢模型。方法本研究采用液相色谱-高分辨质谱技术检测斑...目的随着新型合成大麻素的演变,其在体内代谢速度增快,对原药的检测难度增加,为确定是否吸食合成大麻素,其生物标志物可以作为强有力的证据,常见的代谢模型有体内代谢模型和体外代谢模型。方法本研究采用液相色谱-高分辨质谱技术检测斑马鱼体内代谢模型和人肝微粒体体外代谢模型中MDMB-4en-PICA的代谢情况,并使用Trance Finder 4.1通用版进行数据采集和Compound Discoverer 2.1版本进行代谢物鉴定。结果斑马鱼模型代谢产生26种代谢物,人肝微粒体模型代谢产生17种代谢物,总共32种代谢物,其中包括23种Ⅰ相代谢物和9种Ⅱ相代谢物,涉及10种代谢途径。结论结果显示,两种模型中二氢二醇代谢物(M16)的峰面积均最高,其次是羟基化代谢物(M22),而且具有一定的特异性,可以推荐作为MDMB-4en-PICA潜在的生物标志物。展开更多
Sulfonamides(SAs)are one of the most widely used antibiotics and their residuals in the environment could cause some negative environmental issues.Advanced oxidation such as Fenton-like reaction has been widely applie...Sulfonamides(SAs)are one of the most widely used antibiotics and their residuals in the environment could cause some negative environmental issues.Advanced oxidation such as Fenton-like reaction has been widely applied in the treatment of SAs polluted water.Degradation rates of 95%-99.7%were achieved in this work for the tested 8 SAs,including sulfisomidine,sulfameter(SME),phthalylsulfathiazole,sulfamethoxypyridazine,sulfamonomethoxine,sulfisoxazole,sulfachloropyridazine,and sulfadimethoxine,in the Fe_(3)O_(4)/peroxodisulfate(PDS)oxidation system after the optimization of PDS concentration and p H.Meanwhile,it was found that a lot of unknown oxidation products were formed,which brought up the uncertainty of health risks to the environment,and the identification of these unknown products was critical.Therefore,SME was selected as the model compound,from which the oxidation products were never elucidated,to identify these intermediates/products.With liquid chromatography-high resolution tandem mass spectrometry(LC-HRMS/MS),10 new products were identified,in which 2-amino-5-methoxypyrimidine(AMP)was confirmed by its standard.The investigation of the oxidation process of SME indicated that most of the products were not stable and the degradation pathways were very complicated as multiple reactions,such as oxidation of the amino group,SO_(2)extrusion,and potential cross-reaction occurred simultaneously.Though most of the products were not verified due to the lack of standards,our results could be helpful in the evaluation of the treatment performance of SAs containing wastewater.展开更多
A liquid chromatography-high resolution mass spectrometry （LC-HRMS） method was developed and validated for the determination of piperine （PPR） on dried blood spots （DBS）. DBS samples were prepared by spiking the...A liquid chromatography-high resolution mass spectrometry （LC-HRMS） method was developed and validated for the determination of piperine （PPR） on dried blood spots （DBS）. DBS samples were prepared by spiking the whole blood with analyte to produce 30 μL of blood spots on specimen collection cards. Chromatographic separation was achieved on an Atlantis dC18 column using acetonitrile and water （0.1% formic acid） （85：15, v/v） as mobile phase in an isocratic mode of elution at a flow rate of 0.75 mL/min. MS detection was carried out in electrospray positive ion mode for the target ions and monitored at m/z 286.1465 for PPR and 272.1303 for the internal standard （IS）. The developed method exhibited a linear dynamic range over 0.01-2000 ng/mL for PPR on DBS. The overall extraction recovery of PPR from DBS was 92.5%. Influence of hematocrit and spot volume on DBS was also evaluated and found to be well within the acceptable limits. The method was successfully applied to pharmacokinetic studies of PPR in rats.展开更多
Background: Negative experiences in early life can induce long-lasting effects on the welfare, health, and performance of farm animals. A delayed placement of chicks in rearing houses has negative effects on their per...Background: Negative experiences in early life can induce long-lasting effects on the welfare, health, and performance of farm animals. A delayed placement of chicks in rearing houses has negative effects on their performance, and results in fecal-specific odors detectable by rats. Based on this observation, the volatile organic compounds(VOCs) and metabolites from the feces of 12-day-old chickens were screened for early markers of response to negative events using gas-chromatography and liquid-chromatography coupled with mass spectrometry(GC-MS, LC-HRMS).Results: The low reproducibility of solid-phase micro-extraction of the VOCs followed by GC-MS was not suitable for marker discovery, in contrast to liquid extraction of metabolites from freeze-dried feces followed by GC-MS or LC-HRMS analysis. Therefore, the fecal metabolome from 12-day-old chicks having experienced a normal or delayed placement were recorded by GC-MS and LC-HRMS in two genotypes from two experiments. From both experiments, 25 and 35 metabolites, respectively explaining 81% and 45% of the difference between delayed and control chickens, were identified by orthogonal partial least-squares discriminant analysis from LC-HRMS and GC-MS profiling.Conclusion: The sets of molecules identified will be useful to better understand the chicks’ response to negative events over time and will contribute to define stress or welfare biomarkers.展开更多
文摘Potential mutagenic impurities in Active Pharmaceutical Ingredient, Meropenem Trihydrate were assessed and a novel analytical method for their quantification was developed and validated. This Liquid Chromatographic method using High Resolution Mass Spectrometer (LC-HRMS) technique is proved to be suitable for simultaneous quantification of all ten identified impurities with required specificity, sensitivity, resolution, precision, accuracy, and other method characteristics as per ICH Guidelines. The acceptable limit of less than 2.9 μg/g was considered for evaluations, based on drug substance dosage and duration of treatment. The method stands most sensitive with a Limit of Detection of 0.35 μg/g, considering the challenge full acceptance criteria as per current regulatory standards.
基金Conselho Nacional de Desenvolvimento Cientifico e Tecnologico(CNPq)and Fundacao de Amparo a Pesquisa do Estado de Minas Gerais(FAPEMIG)for financial support and research fellowships.
文摘A simple and high-throughput method to simultaneously determine selected benzodiazepines (i.e., diazepam, lorazepam, clonazepam, and bromazepam) in urine was developed and validated. The entire methodology consisted of the application of an innovative extraction/cleanup procedure, namely liquid-liquid extraction with low-temperature partitioning (LLE-LTP), and analysis by liquid chromatography combined with high-resolution mass spectrometry (LC-HRMS). The LLE-LTP procedure was optimized via factorial design and by evaluating crucial variables, specifically the freezing mode (either slow or fast), the urine/acetonitrile volume ratio, and the sample ionic strength. The benzodiazepines were quantified using matrix-matched calibration curves where the following parameters were assessed by validation protocol: in general, linearity range of 17 - 200 μg?L–1 (r > 0.9957);limits of detection lower than 5 μg?L–1;relative standard deviations (RSD) lower than 12.5%;and accuracy ranging from 72.3 to 117%. To test this procedure’s performance, the method was applied to determine the content of diazepam in actual urine samples. The validation results obtained for the method demonstrated that the present methodology could be potentially applied in proficient laboratories as a routine approach for determining benzodiazepines compounds content in urine.
文摘N-Methyl bis(2-(alkyloxy-alkylphosphoryloxy)ethyl)amines, which are abbreviated as PNPs, are a series of new skeleton chemicals belonging to schedule 2.B.04 chemicals of Chemical Weapons Convention (CWC). PNPs are important markers of chemical warfare agents because they are structurally relative to both nerve agents and N-mustards. In this study, fragmentation pathways of the most characteristic fragment ions in Q-TOF mass spectrometry were proposed based on the information from accurate mass and secondary fragmentations of product ions scan experiments. Results indicated that the base ion in LC/HRMS was the quasi-molecular ion [M+H]+. In LC-HRMS/MS, it was [M+H-CnH2n+1P(O)(OH)CmH2m+1O]+ fragment ion which was formed by losing an alkyloxy alkylphosphoryloxy group from the quasi-molecular ion. The diagnostic ion m/z84.0814 was identified as [C5H10N]+, which was the group of (CH2=CH)2N+(H)CH3. PNPs have two protonated centers. One is on the N atom, the other is on the O atom (P=O). O-n-propyl PNPs generally exhibited two fragmentation pathways. Firstly, the quasi-molecular ion [M+H]+ lost a propoxy alkylphosphoryloxy group to produce [R1P(OH+)(O-n-C3H7)OCH2CH2N(CH3)CH=CH2]+, which could be fragmented further to produce [C5H10N]+ ion. Secondly, [R1P(OH+)(O-n-C3H7) OCH=CH2]+ ions were produced from [M+H]+ and fragmented further to produce the abundant ions [R1P(OH+)(OH)OCH =CH2]. However, O-isopropyl PNPs characteristically produced weak fragment ions [M+H-C3H6]+, which were presumably formed via loss of CH3CH=CH2 from [M+H]+. Other PNPs showed similar fragmentation pathways as O-n-propyl PNPs. On the summarization of the MS fragmentation pathways of PNPs, LC-HRMS/MS quantitative and qualitative methods were developed and applied to analyze N-Methyl bis(2-(butoxy-methylphosphoryloxy)ethyl]amine in high background organic samples. The analytical results had successfully supported the sample preparation for the 33rd official proficiency test of Organization for Prohibition of Chemical Weapons (OPCW).
基金financially supported by the Ministry of Business,Innovation and Employment(MBIE)through LASRA Strategic Science Investment Funding(SSIF).Grant Number LSRX1801.
文摘Collagen structure in biological tissues imparts its intrinsic physical properties by the formation of several covalent crosslinks.For the first time,two major crosslinks in the skin dihydroxylysinonorleucine(HLNL)and histidinohydroxymerodesmosine(HHMD),were isotopically labelled and then analysed by liquid-chromatography high-resolution accurate-mass mass spectrometry(LC-HRMS)and small-angle neutron scattering(SANS).The isotopic labelling followed by LC-HRMS confirmed the presence of one imino group in both HLNL and HHMD,making them more susceptible to degrade at low pH.The structural changes in collagen due to extreme changes in the pH and chrome tanning were highlighted by the SANS contrast variation between isotopic labelled and unlabelled crosslinks.This provided a better understanding of the interaction of natural crosslinks with the chromium sulphate in collagen suggesting that the development of a benign crosslinking method can help retain the intrinsic physical properties of the leather.This analytical method can also be applied to study artificial crosslinking in other collagenous tissues for biomedical applications.
文摘目的随着新型合成大麻素的演变,其在体内代谢速度增快,对原药的检测难度增加,为确定是否吸食合成大麻素,其生物标志物可以作为强有力的证据,常见的代谢模型有体内代谢模型和体外代谢模型。方法本研究采用液相色谱-高分辨质谱技术检测斑马鱼体内代谢模型和人肝微粒体体外代谢模型中MDMB-4en-PICA的代谢情况,并使用Trance Finder 4.1通用版进行数据采集和Compound Discoverer 2.1版本进行代谢物鉴定。结果斑马鱼模型代谢产生26种代谢物,人肝微粒体模型代谢产生17种代谢物,总共32种代谢物,其中包括23种Ⅰ相代谢物和9种Ⅱ相代谢物,涉及10种代谢途径。结论结果显示,两种模型中二氢二醇代谢物(M16)的峰面积均最高,其次是羟基化代谢物(M22),而且具有一定的特异性,可以推荐作为MDMB-4en-PICA潜在的生物标志物。
基金supported by the National Natural Science Foundation of China(Nos.21607020,21876022,41977197)。
文摘Sulfonamides(SAs)are one of the most widely used antibiotics and their residuals in the environment could cause some negative environmental issues.Advanced oxidation such as Fenton-like reaction has been widely applied in the treatment of SAs polluted water.Degradation rates of 95%-99.7%were achieved in this work for the tested 8 SAs,including sulfisomidine,sulfameter(SME),phthalylsulfathiazole,sulfamethoxypyridazine,sulfamonomethoxine,sulfisoxazole,sulfachloropyridazine,and sulfadimethoxine,in the Fe_(3)O_(4)/peroxodisulfate(PDS)oxidation system after the optimization of PDS concentration and p H.Meanwhile,it was found that a lot of unknown oxidation products were formed,which brought up the uncertainty of health risks to the environment,and the identification of these unknown products was critical.Therefore,SME was selected as the model compound,from which the oxidation products were never elucidated,to identify these intermediates/products.With liquid chromatography-high resolution tandem mass spectrometry(LC-HRMS/MS),10 new products were identified,in which 2-amino-5-methoxypyrimidine(AMP)was confirmed by its standard.The investigation of the oxidation process of SME indicated that most of the products were not stable and the degradation pathways were very complicated as multiple reactions,such as oxidation of the amino group,SO_(2)extrusion,and potential cross-reaction occurred simultaneously.Though most of the products were not verified due to the lack of standards,our results could be helpful in the evaluation of the treatment performance of SAs containing wastewater.
文摘A liquid chromatography-high resolution mass spectrometry （LC-HRMS） method was developed and validated for the determination of piperine （PPR） on dried blood spots （DBS）. DBS samples were prepared by spiking the whole blood with analyte to produce 30 μL of blood spots on specimen collection cards. Chromatographic separation was achieved on an Atlantis dC18 column using acetonitrile and water （0.1% formic acid） （85：15, v/v） as mobile phase in an isocratic mode of elution at a flow rate of 0.75 mL/min. MS detection was carried out in electrospray positive ion mode for the target ions and monitored at m/z 286.1465 for PPR and 272.1303 for the internal standard （IS）. The developed method exhibited a linear dynamic range over 0.01-2000 ng/mL for PPR on DBS. The overall extraction recovery of PPR from DBS was 92.5%. Influence of hematocrit and spot volume on DBS was also evaluated and found to be well within the acceptable limits. The method was successfully applied to pharmacokinetic studies of PPR in rats.
基金supported financially by a “crédits incitatifs” grant from the department of Animal Physiology and Livestock Systems(PHASE)at INRAa grant from the Integrated Management of Animal Health metaprogram of INRA for the “GISA-WHELP” project(www.gisa.inra.fr/en)
文摘Background: Negative experiences in early life can induce long-lasting effects on the welfare, health, and performance of farm animals. A delayed placement of chicks in rearing houses has negative effects on their performance, and results in fecal-specific odors detectable by rats. Based on this observation, the volatile organic compounds(VOCs) and metabolites from the feces of 12-day-old chickens were screened for early markers of response to negative events using gas-chromatography and liquid-chromatography coupled with mass spectrometry(GC-MS, LC-HRMS).Results: The low reproducibility of solid-phase micro-extraction of the VOCs followed by GC-MS was not suitable for marker discovery, in contrast to liquid extraction of metabolites from freeze-dried feces followed by GC-MS or LC-HRMS analysis. Therefore, the fecal metabolome from 12-day-old chicks having experienced a normal or delayed placement were recorded by GC-MS and LC-HRMS in two genotypes from two experiments. From both experiments, 25 and 35 metabolites, respectively explaining 81% and 45% of the difference between delayed and control chickens, were identified by orthogonal partial least-squares discriminant analysis from LC-HRMS and GC-MS profiling.Conclusion: The sets of molecules identified will be useful to better understand the chicks’ response to negative events over time and will contribute to define stress or welfare biomarkers.
