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1.
    
Rosmarinic acid (RA) is a caffeic acid derivative and one of the most abundant and bioactive constituents in Java tea (Orthosiphon stamineus), which has significant biological activities. However, relatively few studies have been conducted to describe this compound's metabolites in vivo. Therefore, an ultra‐high‐performance liquid chromatography coupled to quadrupole‐time‐of‐flight tandem mass spectrometry (UHPLC–QTOF–MS/MS) analysis with a three‐step data mining strategy was established for the metabolic profile of RA. Firstly, the exogenously sourced ions were filtered out by the MarkerView software and incorporated with Microsoft Office Excel software. Secondly, a novel modified mass detects filter strategy based on the predicted metabolites was developed for screening the target ions with narrow, well‐defined mass detection ranges. Thirdly, the diagnostic product ions and neutral loss filtering strategy were applied for the rapid identification of the metabolites. Finally, a total of 16 metabolites were reasonably identified in urine, bile and feces, while metabolites were barely found in plasma. The metabolites of RA could also be distributed rapidly in liver and kidney. Glucuronidation, methylation and sulfation were the primary metabolic pathways of RA. The present findings might provide the theoretical basis for evaluating the biological activities of RA and its future application.  相似文献   

2.
    
Tilianin is an active flavonoid glycoside found in many medical plants. Data are lacking regarding its pharmacokinetics and disposition in vivo. The objective of this study was to develop a sensitive, reliable and validated ultra‐high‐performance liquid chromatography with tandem mass spectrometry (UHPLC–MS/MS) method to simultaneously quantify tilianin and its main metabolites and to determine its pharmacokinetics in wild‐type and breast cancer resistance protein knockout (Bcrp1−/−) FVB mice. Chromatographic separation was accomplished on a C18 column by utilizing acetonitrile and 0.5 mm ammonium acetate as the mobile phase. Mass spectrometric detection was performed using electrospray ionization in both positive and negative modes. The results showed that the precision, accuracy and recovery, as well as the stability of tilianin and its metabolites in mouse plasma, were all within acceptable limits. Acacetin‐7‐glucuronide and acacetin‐7‐sulfate were the major metabolites of tilianin in mouse plasma. Moreover, systemic exposure of acacetin‐7‐sulfate was significantly higher in Bcrp1 (−/−) FVB mice compared with wild‐type FVB mice. In conclusion, the fully validated UHPLC–MS/MS method was sensitive, reliable, and was successfully applied to assess the pharmacokinetics of tilianin in wild‐type and Bcrp1 (−/−) FVB mice. Breast cancer resistance protein had a significant impact on the elimination of the sulfated metabolite of tilianin in vivo.  相似文献   

3.
    
Carbenoxolone is a derivative of glycyrrhetinic acid found in the root of Glycyrrhiza glabra, colloquially known as licorice. It has been used as a treatment for peptic and oral ulcers. In recent years, carbenoxolone has been utilized in basic research for its ability to block gap junctional communication. Better understanding the distribution of carbenoxolone after systemic administration can lead to a better understanding of its potential sites of action. Presented is an ultra high‐performance liquid chromatography tandem mass spectrometer (UHPLC–MS/MS) method for the identification and quantification of carbenoxolone in mouse blood and brain tissue. Twenty mice were injected intraperitoneally with 25 mg/kg carbenoxolone and brain tissue and blood were collected for analysis. Blood concentrations (mean ± SD) at 15, 30, 60 and 120 min were determined to be (n = 5) 5394 ± 778, 2636 ± 836, 1564 ± 541 and 846 ± 252 ng/mL, respectively. Brain concentrations (mean ± SD) at 15, 30, 60 and 120 mins were determined to be (n = 5) 171 ± 62, 102 ± 35, 55 ± 10 and 27 ± 9 ng/g, respectively. The analysis of these specimens at the four different time points resulted in blood and brain half‐lives in mice of ~43 and 41 min, respectively. The UHPLC–MS/MS method was determined to be sensitive and robust for quantification of carbenoxolone.  相似文献   

4.
    
Quizartinib is a highly potent inhibitor of the fms‐like tyrosine kinase receptor, which is one of the most commonly mutated genes in acute myeloid leukemia. Quizartinib has shown a significant antileukemic clinical influence among relapsed/refractory acute myeloid leukemia patients. This study aimed at developing and validating an analytical method for the measurement of quizartinib in rat plasma using liquid chromatography–tandem mass spectrometry (LC–MS/MS). The method was validated according to US Food and Drug Administration guidelines, and the results obtained in this work met the set criteria. Liquid–liquid extraction was used and chromatographic separation was achieved on a BEHTM C18 column. Detection of quizartinib was achieved in multiple reaction monitoring mode using positive‐ion mode electrospray ionization. The MS/MS ion transitions at mass‐to‐charge ratios (m/z) of 561.129/114.09 and 441.16/84.03 were monitored for quizartinib and ibrutinib, respectively. The linear detection range was 2–1000 ng/mL (r > 0.998), with intra‐ and inter‐day assay precisions ≤13.07 and 13.17%, respectively. This rapid, simple and sensitive method was validated and successfully applied to the pharmacokinetic study of quizartinib in rat samples.  相似文献   

5.
    
Farrerol is a 2,3‐dihydro‐flavonoid isolated from rhododendron. In this study, a sensitive and selective ultra‐high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method was developed for the determination of farrerol in rat plasma. Liquid–liquid extraction by ethyl ether was used for sample preparation. Chromatographic separation was achieved on an Agilent UHPLC XDB‐C18 column (2.1 × 100 mm, 1.8 μm) with water and methanol (30:70, v /v) as the mobile phase. An electrospray source was applied and operated in negative ion mode; selection reaction monitoring was used for quantification using target fragment ions m/z 299 → 179 for farrerol and m/z 267 → 252 for internal standard. Calibration plots were linear in the range of 2.88–1440 ng/mL for farrerol in rat plasma. Intra‐ and inter‐day precisions were <11.6%, and the accuracy ranged from −13.9 to 11.9%. The UHPLC–MS/MS method was successfully applied in pharmacokinetics and bioavailability studies of farrerol in rats.  相似文献   

6.
Silodosin (SLD) is a novel α1‐adrenoceptor antagonist which has shown promising clinical efficacy and safety in patients with benign prostatic hyperplasia (BPH). However, lack of information about metabolism of SLD prompted us to investigate metabolic fate of SLD in rats. To identify in vivo metabolites of SLD, urine, feces and plasma were collected from Sprague–Dawley rats after its oral administration. The samples were prepared using an optimized sample preparation approach involving protein precipitation followed by solid‐phase extraction and then subjected to LC/HR‐MS/MS analysis. A total of 13 phase I and six phase II metabolites of SLD have been identified in rat urine which includes hydroxylated, N‐dealkylated, dehydrogenated, oxidative, glucosylated, glucuronide and N‐sulphated metabolites, which are also observed in feces. In plasma, only dehydrogenated, N‐dealkylated and unchanged SLD are observed. The structure elucidation of metabolites was done by fragmentation in MS/MS in combination with HRMS data. The potential toxicity profile of SLD and its metabolites were predicted using TOPKAT software and most of the metabolites were proposed to show a certain degree of skin sensitization and occular irritancy. Copyright © 2016 John Wiley & Sons, Ltd.  相似文献   

7.
    
Celosin A (CA), a natural compound isolated from Celosia argentea L., has been shown significant hepatoprotective effect on AHNP‐induced liver injury. This study described a rapid and sensitive ultra‐high‐pressure liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) assay for determination of CA in rat plasma. Methanol‐mediated precipitation was used for sample pretreatment. Chromatographic separation was achieved on a T3 column with gradient elution using water and acetonitrile as mobile phase. Determination was obtained using an electrospray ionization source in negative selected reaction monitoring mode at the transitions of m/z 793.3 → m/z 661.2 and m/z 955.6 → m/z 793.2 for CA and IS, respectively. The assay was linear over the concentration range 0.25–2500 ng/mL (r > 0.995) with a lowest limit of quantification (LLOQ) of 0.25 ng/mL. The intra‐ and inter‐day precisions (RSD) were 1.65–9.84 and 2.46–13.49%, respectively, while accuracy (RR) ranged from 96.21 to 99.45%, respectively. The recovery ranged from 95.09 to 102.22% and the matrix effect from 98.29 to 100.13%. The analyte was stable under the tested storage conditions. The method has been successfully applied to a preclinical pharmacokinetic study in rats after a single intravenous (2 mg/kg) or oral (50 mg/kg) administration. The oral bioavailability of CA was ~1.94%; in addition, there was no difference between male and female rats. This is the first time of the use of an UHPLC–MS/MS method for determination of CA concentration in rat plasma and for evaluation of its pharmacokinetic behavior.  相似文献   

8.
A simple, rapid, and selective method for determination of plasma biotin was developed using ultra‐high‐performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS). After single‐step protein precipitation with methanol, biotin and stable isotope‐labeled biotin as an internal standard (IS) were chromatographed on a pentafluorophenyl stationary‐phase column (2.1 × 100 mm, 2.7 μm) under isocratic conditions using 10 mm ammonium formate–acetonitrile (93:7, v/v) at a flow rate of 0.6 mL/min. The total chromatographic runtime was 5 min for each injection. Detection was performed in a positive electrospray ionization mode by monitoring selected ion transitions at m/z 245.1/227.0 and 249.1/231.0 for biotin and the IS, respectively. The calibration curve was linear in the range of 0.05–2 ng/mL using 300 μL of plasma. The intra‐ and inter‐day precisions were all <7.1%. The accuracy varied from ?0.7 to 8.2%. The developed UHPLC–MS/MS method was successfully applied to determine plasma biotin concentrations in hemodialysis patients. Copyright © 2016 John Wiley & Sons, Ltd.  相似文献   

9.
    
Omarigliptin is a novel long‐acting dipeptidyl peptidase‐4 inhibitor used for the treatment of type 2 diabetes. In this work, a sensitive and selective ultra‐high pressure liquid chromatography tandem mass spectrometry method was developed and validated for the determination of omarigliptin in rat plasma. Sample preparation was performed by protein precipitation with acetonitrile. Chromatographic separation of analytes was achieved on an RRHD Eclipse Plus C18 column (2.1 × 50 mm, 1.8 μm), using gradient mobile phase (0.1% formic acid–acetonitrile) at a flow rate of 0.4 mL/min. Detection was performed in multiple reaction monitoring mode, with target fragment ions m/z 399.1 → 152.9 for omarigliptin and m/z 237.1 → 194 for the internal standard. The total run time was 4 min. Retention time of omarigliptin and internal standard was 1.25 and 2.12 min, respectively. Relative standard deviation (%) of the intra‐ and inter‐day precision was below 10.0%, and accuracy was between 97.9% and 105.3%. Calibration curve was established over the range 2–5000 ng/mL with good linearity. The lower limit of quantification and limit of detection of omarigliptin were 2 and 0.25 ng/mL, respectively. Mean recoveries were in the range 87.3–95.1% for omarigliptin. No matrix effect was observed in this method. This novel method has been successfully applied to a pharmacokinetic study of omarigliptin in rats. The absolute bioavailability of omarigliptin was identified as high as 87.31%.  相似文献   

10.
    
In previous studies, it was revealed that ethyl acetate (EtOAc) extracts from Sophora flavescens Ait. improved glucose tolerance, reduced hyperglycemia, and restored insulin levels in diabetic patients. The aim of this study was to develop an accurate and sensitive UHPLC–MS method for simultaneous determination of flavonoids in EtOAc extracts of Kushen in rat plasma. Ethyl acetate–acetonitrile (2:1) was selected as the solvent to extract the four flavonoids from rat plasma. A BEH C18 column (2.1 mm × 100 mm, 1.7 μm) with a C18 guard cartridge was chosen as the separation plant using a gradient elution with acetonitrile (solvent A) and 0.1% formic acid (solvent B) in water. For all four analytes, the method showed good linearity (r2 > 0.991) in 1–500 ng/mL. The inter‐ and intra‐day accuracy ranged from ?13.78 to 7.19%, and the precision (RSD) was <8.75%. Recoveries of all four flavonoids ranged from 85.9 to 101.3%. According to the results of multitarget pharmacokinetic studies, four active flavonoids in EtOAc extracts from Kushen have similar absorption kinetics but very different metabolic kinetics, and a double peak phenomenon was observed in the concentration–time curve of norkurarinone, which is different from the previous study. In conclusion, detection and multitarget pharmacokinetic studies successfully determined active flavonoids after oral administration of EtOAc extracts from Kushen by an efficient, sensitive and selective UHPLC–MS method, and the results may provide a foundation for future studies of Kushen.  相似文献   

11.
    
A sensitive and reliable ultra‐high‐performance liquid chromatography with tandem mass spectrometry (UHPLC–MS/MS) method was developed and validated for simultaneous determination of l ‐tetrahydropalmatine (l ‐THP) and its active metabolites l ‐isocorypalmine (l ‐ICP) and L ‐corydalmine (l ‐CD) in rat plasma. The analytes were extracted by liquid–liquid extraction and separated on a Bonshell ASB C18 column (2.1 × 100 mm; 2.7 μm; Agela) using acetonitrile–formic acid aqueous as mobile phase at a flow rate of 0.2 mL/min in gradient mode. The method was validated over the concentration range of 4.00–2500 ng/mL for l ‐THP, 0.400–250 ng/mL for l ‐ICP and 1.00–625 ng/mL for l ‐CD. Intra‐ and inter‐day accuracy and precision were within the acceptable limits of <15% at all concentrations. Correlation coefficients (r ) for the calibration curves were >0.99 for all analytes. The quantitative method was successfully applied for simultaneous determination of l ‐THP and its active metabolites in a pharmacokinetic study after oral administration with l ‐THP at a dose of 15 mg/kg to rats.  相似文献   

12.
    
In the present study, a simple, rapid and reliable ultrahigh‐performance liquid chromatography–tandem mass spectrometric (UHPLC–MS/MS) method was developed and validated to determine simultaneously epalrestat (EPA) and puerarin (PUE) in rat plasma for evaluation of the pharmacokinetic interaction of these two drugs. Both the analytes and glipizide (internal standard, IS) were extracted using a protein precipitation method. The separation was performed on a C18 reversed phase column using acetonitrile and 5 mmol/L ammonium acetate in water as the mobile phase with a gradient elution program. The analytes, including IS, were quantified with multiple reaction monitoring under negative ionization mode. The optimized mass transition ion pairs (m /z ) were 318.1 → 274.0 for EPA, 415.1 → 266.9 for PUE and 444.2 → 166.9 for IS. The linear calibration curves for EPA and PUE were obtained in the concentration ranges of 10–4167 and 20–8333 ng/mL, respectively (r > 0.99). The current method was successfully applied for the pharmacokinetic interaction study in rats following administration of EPA and PUE alone or co‐administration (EPA 15 mg/kg, oral; PUE 30 mg/kg, intravenous). The results showed that the combination of EPA and PUE could increase t 1/2 of EPA and reduce T max of EPA. These changes indicated that EPA and PUE might cause drug–drug interactions when co‐administrated.  相似文献   

13.
    
Carboxylesterase and UDP‐glucuronosyltransferase‐mediated metabolism of irinotecan (CPT‐11) has long been proposed to be responsible for its anti‐tumor activity and toxicity, like delayed‐onset diarrhea. However, recent studies failed to gain more comprehensive in vivo and in vitro pharmacokinetic profiles of irinotecan. Herein, we use rat plasma, human liver microsomes and immortalized HepG2 cell as experimental subjects to describe a sensitive and versatile UHPLC–MS/MS method for simultaneously quantifying CPT‐11 and its metabolites, including SN‐38 and SN‐38G. The method was applied to investigate the pharmacokinetic and metabolic behavior of CPT‐11 in the biological samples. Calibration curves for all bio‐matrices showed acceptable linearity (r2 > 0.99). The intra‐ and inter‐day precisions (RSD, %) were within 15% and the excellent accuracy (RE) was between 2.96 and 14.12%. In addition, the specificity, matrix effect and extraction recovery all met the requirements of biological sample analysis. We successfully applied this method to investigate the pharmacokinetics of irinotecan in various biological samples, mediated by carboxylesterase and UDP‐glucuronosyltransferase. This method could be employed in monitoring the metabolic status and clinical efficacy of irinotecan in the future.  相似文献   

14.
Tinoridine is a nonsteroidal anti‐inflammatory drug and also has potent radical scavenger and antiperoxidative activity. However, metabolism of tinoridine has not been thoroughly investigated. To identify in vivo metabolites, the drug was administered to Sprague–Dawley rats (n = 5) at a dose of 20 mg kg?1, and blood, urine and feces were collected at different time points up to 24 h. In vitro metabolism was delved by incubating the drug with rat liver microsomes and human liver microsomes. The metabolites were enriched by optimized sample preparation involving protein precipitation using acetonitrile, followed by solid‐phase extraction. Data processes were carried out using multiple mass defects filters to eliminate false‐positive ions. A total of 11 metabolites have been identified in urine samples including hydroxyl, dealkylated, acetylated and glucuronide metabolites; among them, some were also observed in plasma and feces samples. Only two major metabolites were formed using liver microsomal incubations. These metabolites were also observed in vivo. All the 11 metabolites, which are hitherto unknown and novel, were characterized by using ultrahigh‐performance liquid chromatography–quadrupole time‐of‐flight tandem mass spectrometry in combination with accurate mass measurements. Finally, in silico toxicological screening of all metabolites was evaluated, and two metabolites were proposed to show a certain degree of lung or liver toxicity. Copyright © 2015 John Wiley & Sons, Ltd.  相似文献   

15.
    
To investigate the characteristic of neuromyelitis optica spectrum disorder (NMOSD) in plasma and cerebrospinal fluid, UHPLC–MS was used to identify metabolites by metabolite extraction and on‐machine detection. Multivariate analysis methods were used to complete differential metabolite screening and metabolic pathway analysis. The content of eight substances, such as tryptophan and l ‐glutamic acid in the plasma of NMOSD patients, was higher than that of the healthy control group. Moreover, no differential metabolite was found in the plasma of patients with AQP‐4 antibody‐positive and antibody‐negative NMOSD. The content of five substances including 3‐hydroxybutyric acid in cerebrospinal fluid of patients with NMOSD was reduced. We demonstrated that the distribution of metabolites in plasma between NMOSD patients and healthy counterparts was significantly different. However, there is no significant difference in plasma metabolites between AQP‐4 antibody positive and negative NMOSD. There were some differences in metabolites between the cerebrospinal fluid of NMOSD patients and that of healthy controls. A variety of amino‐acid abnormalities, sphingomyelin dysfunction, energy metabolism and mitochondrial dysfunction are involved in the pathogenesis of NMOSD.  相似文献   

16.
    
A simple, sensitive and specific UHPLC–MS/MS method for quantification of plantagoguanidinic acid (PGA) in rat plasma was applied to investigate the pharmacokinetic behavior in vivo , using protopine as internal standard. The chromatography was separated on a Phenomenex® Luna‐C18 column (2.1 × 150 mm, 3.0 μm) within 7.0 min using a mobile phase consisting of acetonitrile–0.1% formic acid solution under gradient elution at a flow rate of 0.4 mL/min. Prepared samples were monitored by multiple reaction monitoring mode, with the target fragmentions m/z 226.2 → 84.2 for PGA and m/z 354.2 → 188.9 for IS in positive electrospray ionization. The calibration curve of PGA was linear throughout the range 1–1000 ng/mL (r = 0.9962). The lower limit of quantitation in plasma for PGA was 0.1 ng/mL, and the recovery was >88.6%. Intra‐ and interday accuracy ranged from −8.6 to 4.9%. Furthermore, this validated method was successfully used for a pre‐clinical pharmacokinetic study of PGA at a single dose of 20 and 5 mg/kg in rats via oral and intravenous administration. The study showed that PGA was absorpted rapidly and eliminated gradually with a greater absolute oral bioavailability of 70.1% in rats.  相似文献   

17.
    
Traditional herbal medicine consists of multiple components. There are interactions among the components, which affect both potency and toxicity. The preparation of herbal medicines can be a cause of interactions between multicomponents in herbs. To demonstrate the differences in multiherb interactions based on the preparation methods, the changes in the active components in the different preparations of Socheongryong‐tang (SCRT) were evaluated using metabolomics profiling. We performed multicomponent profiling of the decoction of SCRT (SCRTD) and individual herb mixture (SCRTM) using ultra‐high‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry (UPLC–QTOF–MS). Active compounds from SCRTD and SCRTM were identified using multivariate analysis, and the activities between the two groups were compared. We also evaluated the anti‐inflammatory effect of SCRT through investigating the protein expression of iNOS and COX‐2 in lipopolysaccharide‐induced macrophage RAW 264.7 cells in both groups. From the multivariate analysis, 53 active compounds that have different intensities between SCRTD and SCRTM were identified. The intensities of those components, such as ephedrines, glycyrrhizic acid, 6‐gingerol and (2E,4E,8Z,10E)‐N‐isobutyl‐2,4,8,10‐dodecatetraenamide, which is newly identified in Asiasarum heterotropoides, were mostly higher in SCRTD than in SCRTM, which was related to the anti‐inflammatory effect. From the iNOS inhibition test, it was found that SCRTD had a stronger anti‐inflammatory effect than SCRTM. It was demonstrated that multicomponent interactions can be changed by the preparation method, and finally the anti‐inflammatory effect in SCRT can be affected.  相似文献   

18.
    
Lomatogonium rotatum (L.) Fries ex Nym (L. rotatum), a member of Gentianaceae, is an important mongolian medicine in China used to treat febrile diseases in liver and gallbladder. The aim of present study was to investigate the chemical constituents and metabolites of the 50% ethanol fraction of L. rotatum (50EtLR). Firstly, the extract of L. rotatum was partitioned by macroporous resin to obtain the target fraction (50EtLR), then several compounds were isolated from 50EtLR to obtained the standards for further analysis of chemical constituents of 50EtLR. Secondly, the chemical constituents of 50EtLR were characterized using the ultra‐high performance liquid chromatography coupled with quadrupole–time‐of‐flight mass spectrometry (UHPLC–Q‐TOF–MS/MS). Finally, prototype constituents and related metabolites were analyzed after orally administerng 50EtLR to rats. As a result, a new compound, 6‐O‐[β‐d ‐xylopyranosyl‐(1 → 6)‐O‐β‐d ‐glucopyranosyl]‐1,4,8‐trimethoxyxanthone ( 6 ) along with seven known compounds ( 1–5 , 7 and 8 ) were isolated from the 50EtLR, 92 components were either unambiguously or tentatively identified. Additionally, 34 prototype constituents and 112 metabolites in rat plasma along with 32 prototype constituents and 53 metabolites in rat liver were tentatively identified. Therefore, xanthones and flavonoids were the main chemical constituents of 50EtLR and sulfation and glucuronidation are the main enzyme‐induced metabolic pathways involved post‐administration.  相似文献   

19.
    
A fully valid UHPLC–MS/MS method was developed for the determination of etoposide, gemcitabine, vinorelbine and their metabolites (etoposide catechol, 2′,2′‐difluorodeoxyuridine and 4‐O ‐deacetylvinorelbine) in human plasma. The multiple reaction monitoring mode was performed with an electrospray ionization interface operating in both the positive and negative ion modes per compound. The method required only 100 μL plasma with a one‐step simple de‐proteinization procedure, and a short run time of 7.5 min per sample. A Waters ACQUITY UPLC HSS T3 column (2.1 × 100 mm, 1.8 μm) provided chromatographic separation of analytes using a binary mobile phase gradient (A, 0.1% formic acid in acetonitrile, v /v; B, 0.1% formic acid in water, v /v). Linear coefficients of correlation were >0.995 for all analytes. The relative deviation of this method was <10% for intra‐ and inter‐day assays and the accuracy ranged between 86.35% and 113.44%. The mean extraction recovery and matrix effect of all the analytes were 62.07–105.46% and 93.67–105.87%, respectively. This method was successfully applied to clinical samples from patients with lung cancer.  相似文献   

20.
    
Plasma concentrations of nicotine and its active metabolite cotinine are highly correlated with its biological effects. A UHPLC–MS/MS method was developed, validated and applied for nicotine and cotinine analysis in mice plasma. Chromatographic separation was achieved on a BEH HILIC column using acetonitrile (0.1% formic acid) and 10 mm ammonium formate as mobile phase. The gradient elution was performed at 0.4 mL/min with a run time of 3.6 min. The quantitative ion transition was m/z 163.1 > 130.0 for nicotine, m/z 177.1 > 80.0 for cotinine and m/z 167.1 > 134.0 for nicotine‐D4 (internal standard, IS). For both nicotine and cotinine, the calibration range was 5–500 ng/mL with 5 ng/mL as the lower limit of quantitation, and the intra‐ and inter‐day bias and imprecision were ?4.61–12.00% and <11.12%. The IS normalized recovery was 90.62–98.95% for nicotine and 89.18–101.53% for cotinine, and the IS normalized matrix factor was 106.00–116.44% for nicotine and 100.34–109.85% for cotinine. Both nicotine and cotinine were stable under conventional storage conditions. The validated method has been applied to a pharmacokinetic study in mice to calculate the pharmacokinetic parameters for both analytes.  相似文献   

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