Identification of components and metabolites of traditional Chinese medicines(TCMs)employing liquid chromatography-quadrupole time-of-flight mass spectrometry(LC-Q-TOF MS)techniques with information-dependent acquisit...Identification of components and metabolites of traditional Chinese medicines(TCMs)employing liquid chromatography-quadrupole time-of-flight mass spectrometry(LC-Q-TOF MS)techniques with information-dependent acquisition(IDA)approaches is increasingly frequent.A current drawback of IDA-MS is that the complexity of a sample might prevent important compounds from being triggered in IDA settings.Sequential window acquisition of all theoretical fragment-ion spectra(SWATH)is a dataindependent acquisition(DIA)method where the instrument deterministically fragments all precursor ions within the predefined m/z range in a systematic and unbiased fashion.Herein,the superiority of SWATH on the detection of TCMs’components was firstly investigated by comparing the detection efficiency of SWATH-MS and IDA-MS data acquisition modes,and sanguisorbin extract was used as a mode TCM.After optimizing the setting parameters of SWATH,rolling collision energy(CE)and variable Q1 isolation windows were found to be more efficient for sanguisorbin identification than the fixed CE and fixed Q1 isolation window.More importantly,the qualitative efficiency of SWATH-MS on sanguisorbins was found significantly higher than that of IDA-MS data acquisition.In IDA mode,18 kinds of sanguisorbins were detected in sanguisorbin extract.A total of 47 sanguisorbins were detected when SWATH-MS was used under rolling CE and flexible Q1 isolation window modes.Besides,26 metabolites of sanguisorbins were identified in rat plasma,and their metabolic pathways could be deduced as decarbonylation,oxidization,reduction,methylation,and glucuronidation according to their fragmental ions acquired in SWATH-MS mode.Thus,SWATH-MS data acquisition could provide more comprehensive information for the component and metabolite identification for TCMs than IDA-MS.展开更多
Polyoxyethylene glycerol ricinoleate(PGR) serves as a solubilizer/emulsifier that is commonly used in pharmaceutical formulations despite being associated with severe anaphylactoid hypersensitivity reactions.Cremophor...Polyoxyethylene glycerol ricinoleate(PGR) serves as a solubilizer/emulsifier that is commonly used in pharmaceutical formulations despite being associated with severe anaphylactoid hypersensitivity reactions.Cremophor EL?(CrEL) is the most representative PGR produced from reacting ethylene oxide with castor oil.To help clarify the cause of side effects and potentially improve the safety of PGR-based drug delivery vehicle,we have developed separate but related analytical methods for the quantitation of CrEL and its main metabolites,glycerol ethoxylate(GE) and ricinoleic acid(RA).Since CrEL and GE are highly disperse mixtures of polymers that are not amenable to analysis by conventional liquid chromatographytandem mass spectrometry(LC-MS/MS),we used liquid chromatography-triple-quadrupole-time-of-flight mass spectrometry(LC-Q-TOF MS) combined with product ion data acquisition by MSALLand sequential window acquisition of all theoretical fragments mass spectrometry(SWATH MS),respectively to perform the analysis.In contrast,RA is a single molecular entity that could be readily analyzed using conventional LC-HR MS/MS.Selection of specific fragment ions for CrEL,GE,RA and their internal standards enabled a precise quantitation of such a complex analytes system in rat plasma after a single and simple sample preparation method.Assay validation indicated linearity for CrEL,GE and RA over the concentration ranges 0.2~20.0 μg/mL,0.1~10.0 μg/mL and 0.1~20.0 μg/m L,respectively with satisfactory results for other validation parameters.A subsequent pharmacokinetic study involving single intravenous 200 mg/kg injections of CrEL to rats showed the methods enable comprehensive and high throughput quantitation of CrEL and its metabolites in a biological matrix.Our combination of assays provides effective application in investigating the cause of the hypersensitivity reaction of PGR and potentially to improve its safety for using as a vehicle in drug formulations.展开更多
基金the National Natural Science Foundation of China(81573559,81530098)the Ministry of National Science and Technique(Grant No.2017ZX09309027)。
文摘Identification of components and metabolites of traditional Chinese medicines(TCMs)employing liquid chromatography-quadrupole time-of-flight mass spectrometry(LC-Q-TOF MS)techniques with information-dependent acquisition(IDA)approaches is increasingly frequent.A current drawback of IDA-MS is that the complexity of a sample might prevent important compounds from being triggered in IDA settings.Sequential window acquisition of all theoretical fragment-ion spectra(SWATH)is a dataindependent acquisition(DIA)method where the instrument deterministically fragments all precursor ions within the predefined m/z range in a systematic and unbiased fashion.Herein,the superiority of SWATH on the detection of TCMs’components was firstly investigated by comparing the detection efficiency of SWATH-MS and IDA-MS data acquisition modes,and sanguisorbin extract was used as a mode TCM.After optimizing the setting parameters of SWATH,rolling collision energy(CE)and variable Q1 isolation windows were found to be more efficient for sanguisorbin identification than the fixed CE and fixed Q1 isolation window.More importantly,the qualitative efficiency of SWATH-MS on sanguisorbins was found significantly higher than that of IDA-MS data acquisition.In IDA mode,18 kinds of sanguisorbins were detected in sanguisorbin extract.A total of 47 sanguisorbins were detected when SWATH-MS was used under rolling CE and flexible Q1 isolation window modes.Besides,26 metabolites of sanguisorbins were identified in rat plasma,and their metabolic pathways could be deduced as decarbonylation,oxidization,reduction,methylation,and glucuronidation according to their fragmental ions acquired in SWATH-MS mode.Thus,SWATH-MS data acquisition could provide more comprehensive information for the component and metabolite identification for TCMs than IDA-MS.
基金supported by grants from the National Natural Science Foundation of China (Nos.81872831 and 82030107)the National Science and Technology Major Projects for “significant new drugs creation” of the 13th five-year plan (Nos.2017ZX09101001 and 2018ZX09721002007, China)。
文摘Polyoxyethylene glycerol ricinoleate(PGR) serves as a solubilizer/emulsifier that is commonly used in pharmaceutical formulations despite being associated with severe anaphylactoid hypersensitivity reactions.Cremophor EL?(CrEL) is the most representative PGR produced from reacting ethylene oxide with castor oil.To help clarify the cause of side effects and potentially improve the safety of PGR-based drug delivery vehicle,we have developed separate but related analytical methods for the quantitation of CrEL and its main metabolites,glycerol ethoxylate(GE) and ricinoleic acid(RA).Since CrEL and GE are highly disperse mixtures of polymers that are not amenable to analysis by conventional liquid chromatographytandem mass spectrometry(LC-MS/MS),we used liquid chromatography-triple-quadrupole-time-of-flight mass spectrometry(LC-Q-TOF MS) combined with product ion data acquisition by MSALLand sequential window acquisition of all theoretical fragments mass spectrometry(SWATH MS),respectively to perform the analysis.In contrast,RA is a single molecular entity that could be readily analyzed using conventional LC-HR MS/MS.Selection of specific fragment ions for CrEL,GE,RA and their internal standards enabled a precise quantitation of such a complex analytes system in rat plasma after a single and simple sample preparation method.Assay validation indicated linearity for CrEL,GE and RA over the concentration ranges 0.2~20.0 μg/mL,0.1~10.0 μg/mL and 0.1~20.0 μg/m L,respectively with satisfactory results for other validation parameters.A subsequent pharmacokinetic study involving single intravenous 200 mg/kg injections of CrEL to rats showed the methods enable comprehensive and high throughput quantitation of CrEL and its metabolites in a biological matrix.Our combination of assays provides effective application in investigating the cause of the hypersensitivity reaction of PGR and potentially to improve its safety for using as a vehicle in drug formulations.
