ABSTRACT
OBJECTIVE: To evaluate the effectiveness and safety of Asarum insigne Diels and develop a sensitive, accurate, and efficient ultra-fast liquid chromatography coupled with triple quadrupole mass spectrometer (UFLC-MS/MS) method for simultaneous quantification of five compounds. METHODS: Chromatographic separation was performed on a Shimadzu shim-pack XR-ODS III column(2.0 mm×75 mm,1.6 μm). The mobile phase was composed of 0.1% aqueous formic acid and methanol and the flow rate was 0.2 mL•min-1. Using the established method, all components could be easily separated within 14 min. All components were detected in multiple reaction monitor mode after positive atmospheric pressure chemical ionization. RESULTS: The method was validated, and had good precision, linearity, lower limit of detection, lower limit of quantification, recovery, and stability. This method was used to determine and compare the contents of five components in different medicinal parts of 11 samples, and the contents of 3,4,5-trimethoxytoluene, 3,5-dimethoxytoluene, methyleugenol in different medicinal parts of Asarum insigne were reported for the first time, which (in ng•mg-1) were as follows: 3,4,5-trimethoxytoluene: 1.5-16.71; 3,5-dimethoxytoluene: 28.59-177.20; methyleugenol: 2.46-22.48; AL-I: 46.39-324.04; 7-OCH3-AL-IV: 12.95-251.04. The content of 7-OCH3-AL-IV and total contents of two aristololactams in roots were lower than those in leaves or flower parts, and the contents of AL-I in flowers (171.9-324.0 ng•mg-1) were higher than those in roots (78.44-124.56 ng•mg-1). For methyleugenol, the contents in roots were lower than those in the other parts. CONCLUSION: It is recommended to remove the leaves, flowers, and rhizomes when using Asarum insigne and use it with extreme caution. The UFLC-MS/MS method established in this study can provide guidance for the quality evaluation and safe clinical use of traditional Chinese medicine derived from Asarum insigne.
ABSTRACT
In this study, we accurately collected the embryonic parenchyma cells and endocarp stone cells of Arctii Fructus at five different growth stages by laser microdissection. Quantitative analyse of caffeic acid, arctiin and arctigenin in these cells were performed using ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS). The results showed that a large amount of arctiin was produced and accumulated in embryonic parenchyma cells from the late flowering stage to mature stage, while much lower content of arctiin was produced and accumulated in endocarp stone cells at these stages. It suggested that the biosynthetic pathways of arctiin were different in embryonic parenchyma cells from endocarp stone cells of Arctii Fructus. Arctigenin was found to be produced and accumulated in both embryonic parenchyma cells and in endocarp stone cells from the late flowering stage to mature stage, but it reached a peak in endocarp stone cells at late flowering stage, then decreased slowly. The concentration of arctigenin was far less than that of arctiin regardless of embryonic parenchyma cells or endocarp stone cells. These results have validated the new method for analysis of dynamic accumulation of arctiin in Arctii Fructus by UFLC-MS/MS with frozen sections and microdissection.
ABSTRACT
Objective To establish a QTRAP-UFLC-MS/MS method for the determination of 15 kinds of amino acids and 10 kinds of nucleosides in the seeds of Annona squamosa from different producing areas, toanalyze and investigate the difference of the 25 components in samples from five habitats, and provide scientific basis for its comprehensive utilization. Methods The analysis was carried out on a Waters XBridge Amide column (100 mm × 2.1 mm, 3.5 μm) with elution by mobile phase of 0.2% formic acid in water (A)-0.2% formic acid in acetonitrile (B) at a flow rate of 0.6 mL/min. The column temperature was maintained at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. The comprehensive evaluation of the seeds from A. squamosa in five habitats was carried out by PCA and TOPSIS analysis. Results All 15 kinds of amino acids and 10 kinds of nucleosides showed good linearity (r2 > 0.994 7). The RSDs of the precision, repeatability, and stability tests were less than 3.87%. The average recovery rates were in the range from 94.90% to 98.78%, RSDs were in the range from 0.84% to 3.01%. There are differences in the composition and content of amino acids and nucleosides in the seeds of A. squamosa from different habitats, among which Hainan produced the highest comprehensive quality. Conclusion The method is simple, accurate, and reliable, which can provide the reliable basis for quality evaluation of the seeds from A. squamosa, and samples from different habitats can be optimized according to the target component.
ABSTRACT
A method of ultra flow liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) was developed to elucidate the impurity of linezolid tablets. Linezolid was subjected to forced degradation under hydrolytic (acid, base and neutral), oxidative, photolytic and thermal. The structure identification of the degradation products and the fragmentation patterns for the related impurities were analyzed. A total of four degradation impurities were characterized, impurity 1 is (S)-1-amino-3-((3-fluoro-4-morpholinophenyl)amino)propan-2-ol, impurity 2 is (S)-4-(4-(5-(acetamidomethyl)-2-oxo-oxazolidin-3-yl)-2-fluorophenyl)morpholine 4-oxide, impurity 3 is (S)-5-(aminomethyl)-3-(3-fluoro-4-morpholinophenyl)oxazolidin-2-one, impurity 4 is (R)-N-(3-((3-fluoro-4-morpholinophenyl)amino)-2-hydroxypropyl)acetamide. Acid degradation induced impurity 3 and impurity 4, base degradation induced impurity 1 and impurity 4, oxidation degradation induced impurity 2, hydrolysis degradation induced impurity 4. The study also determined calibration factor using impurity references, and the calibration factors were found to be 1.3, 1.4, 0.9 and 1.1, respectively. The toxicity of the degradation impurities was predicted by web-based prediction system. The results from this study provide an important reference in quality control and evaluation of linezolid.
ABSTRACT
An effective herbal medicinal prescription of Shengjiang Xiexin decoction (SXD) was used in treating the inflammatory bowel disease in clinic. In this study, an ultrafast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was developed to separate and to simultaneously determine 14 major active ingredients in SXD. Chromatographic separation was successfully accomplished on an Acquity BEH C18 (100 mm×2.1 mm, 1.7 μm) column using gradient elution with 0.1% () formic acid water (A) and 0.1% () formic acid in methanol (B). Negative and positive electrospray ionization tandem mass spectrometry was used to detect the 14 analytes using its selective reaction monitoring (SRM) mode. A good linear regression relationship for each analyte was obtained over the range from 3.88 ng/mL to 4080 ng/mL. The precision was evaluated by intra- and inter-day assays with a relative standard deviation (RSD) of less than 6.25%. The recovery measured at three concentration levels varied from 98.72% to 103.47%. The overall limits of quantification (LOQ) ranged from 2.05 ng/mL to 4.72 ng/mL. The method was successfully implemented in the qualitative and quantitative analyses of the 14 chemical constituents in SXD. The results showed that the developed UFLC-MS/MS method was linear and accurate. The method could be used reliably as a quality control method for SXD.
ABSTRACT
A rapid and accurate method of UFLC-Q-TOF-MS/MS combined with multivariate statistical analysis was established for the identification of Ainsliaea fragrans from different origins in this study. The A. fragrans from different producing areas of Jiangxi, Yunnan, Henan and Jiangsu were determined by UFLC-Q-TOF-MS/MS in the negative ion mode. And the data of the study were analyzed by the Markerview and other software for the PCA and OPLS-DA cluster analysis as well as t test. The results of the principal component analysis(PCA)showed that the main components from different origins were well distinguished. And the results of multivariate statistical showed the differences and similarities between different producing areas. Besides, 40 different compounds were identified in the negative ion mode. This method for identifying A. fragrans from different producing areas has the advantages of rapid accuracy and simplicity, which laid the foundation for the evaluation of the quality of the A. fragrans.
ABSTRACT
To establish a UFLC-MS/MS method for the determination of valproate acid in human plasma. Methods:The sample was precipitated by methanol. The analysis of valproate acid and diclofenac sodium ( internal standard) was carried out on a Shim-pack XR-ODS II C18 column (2. 0 × 75 mm,2. 2 μm). Gradient elution was adopted with acetonitrile and water (containing 5 mmol·L-1 ammonium acetate) at a flow rate of 0. 3 ml·min-1. The detection was performed with multiple reactions monitoring (MRM) using nega-tive electrospray ionization (ESI) at m/z 142. 8→142. 8 for valproate acid and m/z 294. 0→249. 8 for diclofenac sodium. Results: The calibration curve of valproate acid was linear over the range of 8.4-168.0 μg·ml-1(r=0.997 5). Inter- and intra-day RSDs were less than 15%, and the analysis was proven to be stable. Totally 30 samples determined by EMIT were assayed by the method. And the results of the two methods analyzed by independent t-test showed no statistical significance. Conclusion:The method is rapid,sensitive and spe-cific,which can be applied in the determination of valproate acid in human plasma.
ABSTRACT
A rapid and sensitive method based on ultrafast liquid chromatography-tandem mass spectrometry was developed and validated for simultaneous determination of Sudan Ⅰ, Sudan Ⅱ, Sudan Ⅲ, and Sudan Ⅳ levels in rat whole blood. Cleanert C18 mixed-mode polymeric sorbent was used for effective solid-phase extraction cleanup. Separation was carried out on a reversed-phase C18 column (100 mm × 2.1 mm, 1.8 μm) using 0.1% (v/v) formic acid in water/0.1% (v/v) formic acid in acetonitrile as the mobile phase in gradient elution. Quantification was performed by an electrospray ionization source in the positive multiple reaction monitoring mode using D5-Sudan I as the internal standard. Calibration curves showed good linearity between 0.2 and 20.0 μg/L, with correlation coefficients higher than 0.9990. The average recovery rates were between 93.05% and 114.98%. The intra- and inter-day relative standard deviations were within 6.2%. The lower limit of quantification was 0.2 μg/L. All the analytes were found to be stable in aseries of stability studies. The proposed method was successfully applied to a pharmacokinetic study of four Sudan dyes after oral administration to rats.
ABSTRACT
OBJECTIVE: To develop a method for determining the concentration of lamivudine in placental perfusate by employing ultra fast liquid chromatography with tandem mass spectrometry (UFLC-MS/MS). METHODS: Famotidine was used as the internal standard. After protein precipitation with acetonitrile, the chromatography was performed on a Shim-pack XR-ODS II C18 column (2.0 mm × 75 mm, 2.2 μm) with mobile phase consisting of methanol and 10 mmol · L-1 ammonium acetate, gradient elution was used, and the flow rate was 0.4 mL · min-1. Tandem mass spectrometric detection was conducted by using multiple reaction monitoring (MRM) in positive ionization mode with an electrospray ionization interface. RESULTS: The calibration curve of lamivudine was linear in the range of 1-2500 ng · mL-1. The intra- and inter-batch RSDs were less than 10%. The method recovery was more than 72%. CONCLUSION: The method is rapid and accurate, which is suitable for the determination of lamivudine in placental perfusate.
ABSTRACT
OBJECTIVE: To establish a UPLC-MS/MS method for the determination of antipyrine in placental perfusate. METHODS: Sample was precipitated with acetonitrile. Analysis of antipyrine and estazolam(internal standard) were carried out on a Shim-pack XR-ODS II C18 column(2.0 mm × 75 mm, 2.2 μm). Gradient elution was adopted using acetonitrile and water (containing 0.1% formic acid, V/V) at a flow rate of 0.4 mL · min-1. Detection was performed with multiple reactions monitoring (MRM) using positive electrospray ionization(ESI) at m/z 189.2→56.2 for antipyrine and m/z 295.2→205.2 for estazolam. RESULTS: The calibration curve of antipyrine was linear over the range of 1.2 -1200 ng · mL-1 (r =0.9993). Inter- and intra-day RSDs were less than 15%, and the analytes were proven to be stable. CONCLUSION: This method is rapid, sensitive and specific, and can be applied to the determination of antipyrine in placental perfusate.
ABSTRACT
OBJECTIVE: To develop a UFLC-MS/MS method for the determination of simvastatin in human plasma, so as to investigate the relative bioavailability of two kinds of simvastatin tablets. METHODS: After liquid-liquid extraction by methyl tert-butyl ether, the analytes were seperated on a Shim-pack XR-ODS column with a gradient elution by the mobile phase of water (A) and aceto-nitrile (B). Conditions of the gradient elution were as follows: 0.01-3 min, B:80%-90%; 3-3.5 min, B:90%-95%, 3.51-5 min, B:80%. Multiple reaction monitor (MRM) was used to determine the concentration of simvastatin (m/z 441.2 → 325.1) and the internal standard, lovastatin (m/z 427.2 → 325.2). A single oral dose of 40 mg test or reference preparation was given to 20 healthy volunteers in a randomized crossover design. Blood samples were obtained and analyzed by the validated UFLC-MS/MS method, and the pharmacokinetics and bioavailability were evaluated by Phoenix WinNonlin 6.0 software. RESULTS: The calibration curve of simvastatin was linear over 0.1-20 ng · mL-1 (r=0.9954) with the lower limit of quantity of 0.1 ng · mL-1. The absolute recoveries were between 63.9% and 83.8%, the extraction recoveries were 63.9%-83.8%, and inter- and intra-day RSDs were less than 7.1% and 10.0%, respectively. The main pharmacokinetic parameters of simvastatin of the test and reference tablets were as follows: t1/2 were (3.96 ± 1.65) and (3.77 ± 1.75)h; ρmax were (8.3641 ±4.9898) and (8.4399 ± 4.8566) ng · mL-1; tmax were (1.71 ± 1.19) and (174 ± 1.10) h; AUC0-t were (29.74 ± 13.05) and (31.16 ± 13.92) ng · h · mL-1; AUC0-∞ were (32.35 ±14.56) and (33.39 ± 14.55)ng · h · mL-1, respectively. The relative bioavailability of the test to reference tablets was (97.3 ± 24.3)%. CONCLUSION: The two preparations are bioequivalent. The established method is successfully used for the bioequivalence study of simvastatin preparations.
ABSTRACT
Monolithic silica spin column extraction （MonoSpin-SPE） was developed as a simple, sensitive, and eco-friendly pretreatment method which combined with ultra-fast liquid chromatography-mass spectrometry （UFLC-MS） to determine the levels of six phthalate esters, dimethyl- （DMP）, diethyl- （DEP）, dipropyl- [ DPrP], butyl-benzyl- （BBP）, dicyclohexyl- （DcHP）, and di-n-octyl-（DOP） phthalate in physiological saline samples. Under optimized experimental conditions, the method was linear in the following ranges： 0.2- 50 μg/L for DMP, DEP, DPrP, DcHP and DOP; 5- 100 μg/L for BBP. The correlation coefficients （R2 ） were in the range of 0. 9951 - 0. 9995 for all the analytes and the limits of detection （LODs） and limits of quantification （LOQs） were in the ranges of 0.02 - 0.9 μg/L and 0.08 - 2.7μg/L, respectively. The pretreatment process showed good reproducibility with inter-day and intra-day relative standard deviations （RSDs） below 8.5% and 11.2%, respectively. This method was used to determine the levels of six phthalate esters in physiological saline samples and the recoveries ranged from 71.2% to 107.3%. DMP and DEP were found in actual physical saline samples （brand A and brand B）.
ABSTRACT
Monolithic silica spin column extraction (MonoSpin-SPE) was developed as a simple,sensitive,and eco-friendly pretreatment method which combined with ultra-fast liquid chromatography-mass spectrometry (UFLC-MS) to determine the levels of six phthalate esters,dimethyl-(DMP),diethyl-(DEP),dipropyl-[DPrP],butyl-benzyl-(BBP),dicyclohexyl(DcHP),and di-n-octyl-(DOP) phthalate in physiological saline samples.Under optimized experimental conditions,the method was linear in the following ranges:0.2 - 50 μg/L for DMP,DEP,DPrP,DcHP and DOP; 5- 100,μg/L for BBP.The correlation coefficients (R2 ) were in the range of 0.9951- 0.9995 for all the analytes and the limits of detection (LODs) and limits of quantification (LOQs) were in the ranges of 0.02- 0.9 μg/L and 0.08- 2.7 μg/L,respectively.The pretreatment process showed good reproducibility with inter-day and intra-day relative standard deviations (RSDs) below 8.5％ and 11.2％,respectively.This method was used to determine the levels of six phthalate esters in physiological saline samples and the recoveries ranged from 71.2％ to 107.3％.DMP and DEP were found in actual physical saline samples (brand A and brand B).