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Objective@#To establish a method for rapid determination of 47 volatile organic compounds in the air of workplace using portable gas chromatography-mass spectrometer(GC-MS).@*Methods@#The mixed standard gas with different concentration levels was made by using the static gas distribution method with the high purity nitrogen as dilution gas. The samples were injected into the GC-MS by a hand-held probe. Retention time and characteristic ion were used for qualitative analysis,and the internal standard method was usd for quantitation.@*Results@#The 47 poisonous substances were separated and determined well. The linear range of this method was 0.2-16.0 mg/m3,and the relative standard deviation of 45 volatile ovganic compounds was 3.8%-15.8%. The average recovery was 79.3%-119.0%.@*Conclusion@#The method is simple,accurate,sensitive,has good separation effect,short analysis period, can be used for qualitative and quantitative analysis of volatile organic compounds in the workplace, and also supports the rapid identification and detection of occupational hazards.
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Objective To analyze the efficacy of the combination of MALDI-TOF and immunoadsorption to detect new biomarkers for lupus. Methods Twenty lupus patients at active stage (SLE group), 10 SLE patients in remission (SLE control group), 10 RA patients and 10 PSS patients (other rheumatic disease control group) and 20 healthy volunteers (healthy control group) were enrolled. The serum samples before and after immunoadsorption from SLE group and those from the control groups were co-incubated with activated chitosan copper derivative nano material. The adsorbed nano material was spotted onto the matrix used in MALDI-TOF for analysis by the Axima-CFR plus MALDI-TOF mass spectrometer. T-test was used for statistical analysis. Results MALDI-TOF MS screening showed that three potential protein biomarkers of mass-to-charge (m/z) ratio 3136, 3264, 3326 were found to be very specific for lupus patients: All of them were expressed before immunoadsorption in high quantity and none of them could be detected both after immunoadsorption and in all the three control groups. None of them (<10 000) were in the molecular weight range of the biomarkers used nowadays such as auto antibodies and complement (>50 000). Conclusion The combination of MALDI-TOF and immunoadsorption is effective in the detection of new serum protein biomarkers for lupus and it may be helpful in the screening of SLE patients at active stage from healthy people.
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Objective: To establish a liquid chromatography-mass spectrometry (LC-MS) method for determining the concentration of amlodipine besylate in human plasma and to evaluate the bioequivalence of 2 kinds of amlodipine besylate tablets. Methods: Twenty healthy male volunteers were enrolled into a single crossover study. A single dose of the suspension equivalent to 10 mg amlodipine besylate or a reference preparation was given in a crossover way. The plasma concentrations of amlodipine besylate were determined by LC-MS method in the volunteers at different time points; the pharmacokinetic parameters and relative bioavailability were calculated and the bioequivalence of the 2 preparations were evaluated. Results: The pharmacokinetic parameters for experimental and the reference preparations were: Cmax (6.21±1.88) vs (6.03±1.08) ng/ml; AUC0-120 (250.68±52.61) vs (246.14±52.11) ng h/ml; Tmax (6.0±2.3) vs (6.1±2.5) h; t1/2 (40.45±6.68) vs (43.74± 9.05) h, respectively. The linear range of the present method was 0.1-20.0 ng/ml; the lowest detectable concentration of amlodipine besylate was 0.1 ng/ ml. There was no significant difference in pharmacokinetic parameters between the 2 tablets. Conclusion: The present method is simple to use, fast, and accurate. The 2 preparations of amlodipine besylate are bioequivalent.
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Objective To evaluate the diagnostic value of surface enhanced laser desorption/ionization time of flight mass spectrom-etry(SELDI-TOF-MS) for Paget disease. Methods The relative contents of serum proteins of 15 healthy people,15 patients with chronic eczema and 20 patients with Paget were detected by Weak cation exchanger protein chip (WCx2) and SELDI-TOF-MS. Results The con-tents of two proteins (3868Da,8876Da) had significant difference in healthy people and Paget disease patients(P<0.01). 291IDa, 3868Da and 5097Da protein peaks had significant difference in chronic eczema and Paget disease(P<0.05). Conclusion It shows great potentiality for early diagnosis and screening the tumor biomarkers of Paget disease with SELDI-TOF-MS.
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Objective To develop a candidate reference method for the measurement of urea in human serum based on isotope dilution/gas chromatography/mass spectrometry.Methods [13C,15N2]Urea used as internal standard Was added to the serum sample and equilibrated with endogenous nonlabeled urea.The serum samples were treated with anhydrous ethanol to emove proteins by precipitation.The serum urea and labeled urea were converted into a trimethylsilyl derivative of 2-hydroxypyrimidine and analyzed by gas chromatography/mass spectrometry system with selected ion monitoring.The concentration of serum ureaWas calculated by the theory of bracketing method.Results The average value of within-run oefficient of vailation(CV),between-run CV and total CV of the procedure were 0.38%(ranged from 0.12%to 0.47%),O.62%(ranged form 0.49% to 0.87%)and 0.73%(ranged from 0.51% to 0.93%).Respectively.The analytical recoveries ranged from 99.37% to 100.95%.The resuhs of analyzing the certified refefence material SRM909b(Level Ⅰand Ⅱ)showed a bias less than 0.2%.Conclusion The procedure for measuring urea in serum is a highly accurate and precise method and can be used as a candidate reference method for serum urea assays.
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Objective To evaluate the matrix effect of processed materials in serum total glycerol measurement and to assess the accuracy of routine test systems.Methods With an isotope dilution liquid chromatography tandem mass spectrometry method as the comparative method，matrix effects of 28 processed materials on 8 routine test systems were evaluated ccording to the NCCLS EP 14 protocol.The processed materials and 20 flesh patient specimens were analyzed with both the comparative method and each of the evaluated methods and results obtained with the two methods were plotted.Two-tailed 95% prediction intervals for the mean of the flesh patient specimen were computed and results on the processed aterials were compared with these intervals for evaluation of matrix effect.Results with the two methods on fresh samples were also compared for assessment of the accuracy of the routine test systems.Results Some of the processed samples showed matrix effects on some of the routine test systems.The observed matrix effects were system-specific and aused either positive or negative biases.Calibration biases were also observed on some test systems.Conclusion Matrix effect and calibration bias have been observed in serum total glycerol measurements.Continued efforts are needed for improving the accuracy of serum total glycerol measurements.
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Objective To develop a method for measuring serum total glycerol by isotope dilution (ID)liquid chromatography tandem mass spectrometry(LC/MS/MS).Methods Serum samples were mixed with [13C3]-glycerol(internal standard)and treated with alcoholic potassium hydroxide to hydrolyze serum glycerides to glycerol.The natural glycerol and the internal standard were benzoylated with the Schotten-Baumann reaction and the benzoates were analyzed by LC/MS/MS.Results The correlation coefficients between the peak area ratios and glycerol concentrations were 0.999 9 and higher.The withinrun coefficients of variation(CV)for serum total glycerol analysis averaged 0.94%(ranged 0.21%～2.62%)and the total CV 1.15%(0.62%～2.00%).Results on certified reference materials(SRM 909b Level Ⅰ and LevelⅡ;GBW 09146 and GBW 09147)showed an averaged bias of 0.20%(-0.20%～1.06%).Conclusions An ID-LC/MS/MS method for serum total glycerol has been developed.The method iS highly specific and precise and may be used as a candidate reference method for serum total glycerol measurements.
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Objective To investigate the value of detecting hepatitis B virus (HBV) YMDD variants by matrix-assisted laser de-sorption time of flight mass spectrometry (MALDI-TOF MS). Methods The assay is based on polymerase chain reaction (PCR) amplifica-tion and mass measurement of oligonucleotides containing sites of mutation of the YMDD motif. Result The MALDI-TOF MS-based genoty-ping assay was sufficiently sensitive to detect as few as 100 copies of HBV genome per milliliter of serum, and this method had superior spe-cificity for determining mixtures of wild-type and variant viruses. When sera of 40 patients were analyzed, the MALDI-TOF MS-based assay correctly identified known viral variants and additional viral quasi-species not detected by previous methods, as well as their'relative abun-dance. Conclusion The sensitivity, specificity and amenability to high-throughput analysis make MALDI-TOF MS-based assay suitable for mass screening of HBV infected patients who are receiving lamivudine.
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AIM To identify differentially expressed proteins in the plasma that may be used as biomarkers for heroin addiction through a two-dimensional (2-D)gel electrophoresis/mass spectrometry approach. METHODS Following removal of albumin and IgG,the plasma from 5 heroin abusers and 5 normal controls was separated by 2-D gel electrophoresis using immobi-lized pH gradients 4 -7 drystrip and PAGE. Gel ima-ges were analyzed using ImageMaster Elit 5.0. Differ-ential proteins were selected and analyzed through tan-dem mass spectrometry. RESULTS Average number for samples was 350±21 protein spots. In them there were 5 spots that differed by more than 1.5 fold be-tween the two groups obtained through image analysis.Through tandem mass spectrometry above spots were identified as fibrinogen γ (increased by 5 fold), hu-man α1-B-glycoprotein (decreased to 1/1.8 of control group), uncleaved α1-antitrypsin (increased by 2.5 fold), chain of transthyretin (decreased to 1/2 of con-trol group ) and ccruloplasmin (increased by 6.6 fold ). CONCLUSION There are differences between heroin abusers and normal controls in the blood plasma proteome. Some proteins may have a role in the damage to central nervous system through heroin abuse. Such proteins may provide novel biomarkers for diagnosis and therapeutic targeting, as well as clues forunderstanding the mechanism of heroin abuse.
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Objective: To screen differentiated expressed proteins in plasma of ovarian serous cystadenocarcinoma patients by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) associated with bioinformatic support vector machines (SVM) and discuss how to establish algorithmic logical model for diagnosis of ovarian serous cys-tadenocarcinoma and its significance. Methods: SELDI-TOF-MS and CM10 chip were used to analyze the plasma samples from 26 ovarian serous cystadenocarcinoma women and 51 control women including 12 cases of ovarian cyst, 31 cases of uterous leiomyoma, 8 cases of ovarian benign cystadenoma. The data was analyzed by Biomarker Wizard software. The plasma proteomic diagnostic model for ovarian serous cys-tadenocarcinoma patients and control subjects were established by using SVM (a bioinformatic method). Results: Seventy-one differentiated protein peaks were screened by Biomarker Wizard software which were captured by SELDI-TOF-MS from CM10 chip (P <0.01). The proteomic profiling for ovarian serous cystadenocarcinoma was optimized by SVM re-screening. The key m/z value of these 7 proteins was 4 099, 4 477, 4 123, 4 081 and 3 938 (up-regulated), 8 785 and 13 783 (down-regulated). Three-fold cross validation followed by blinded determination demonstrated that the sensitivity and specificity of the established model were 84.62% and 96.08% separately, and the positive predictive value was 92.21% for differential diagnosis of ovarian serous cystadenocarcinoma patients. Conclusion: ProteinChip-mass spectrometry technology can rapidly and effectively screen differentiated proteins from the plasma of ovarian serous cystadenocarcinoma patients. Combined with SVM, a diagnostic model was generated from proteomic patterns of ovarian serous cystadenocarcinoma, which had potential significance for establishing diagnostic methods for ovarian cancer.
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Objective: To compare the proteomic differences in early cervical carcinoma tissues with various radiosensitivities and provide the basis for determining of radiosensitivity-associated proteins in cervical carcinoma. Methods: The fresh carcinoma tissues were collected from 10 untreated cervical carcinoma patients and preserved at - 80C. According to WHO Criterion of Therapeutical Effect of Solid Carcinoma, the tissues were classified into two groups: high sensitive group (HS) and low sensitive group (LS). The total protein was extracted and separated by using two-dimensional gel electrophoresis (2-DE). The PD-quest 7.0 software was used to perform image matching and difference analysis to identify the differentially expressed protein-spots between the two groups. The identified protein spot was cut in electrophoretic gel in situ, digested with trypsin, and analyzed by Matrix-assisted Laser Desorption/ Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS). The peptide mass fingerprintings (PMF) was obtained. The proteins were identified by data searching in the Mascot database. Some differentially expressed proteins were confirmed by Western blotting. Results: The constructed two-dimensional gel electrophoresis profiles had high resolution and good reproducibility for both HS and IS groups. There were (754 + 64) differentially expressed protein spots (n = 5) in HS group and (777 ± 48) in LS group (n = 5). Some protein spots with differential ratio more than 2-fold were selected and analyzed by mass spectrometry. The average matching rate was 87.6% between the two groups. Eight differentially expressed proteins were successfully identified among which five proteins had overexpression and three proteins had weak expression in HS group. The results of Western blotting were consistant with proteomic screening. Conclusion: There was significant difference in protein profilings between HS and LS early cervical carcinoma after radiotherapy. The differentially expressed proteins may be radiosensitivity-associated proteins.
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AIM: To compare the pharmacokinetics and relative bioavailability of the domestic and imported sustained-release tablets of gliclazide in healthy volunteers. METHODS:The study was performed by an four-period crossover design with singledose and multiple-dose administration. The plasmadrug concentrations of twenty male healthy volunteers were determined by liquid chromatography with mass spectrum detector method (LC-MS). RESULTS:The pharmacokinetic parameters after a single oral dose of the domestic and imported gliclazide tablets were (7.2+s 1.5) h and (6.9 +1.4) h for tmax, (13.4 ±1.2) h and (13.7 +1.3) h for t1/2, (2.4 +0.8) mg ·L-1and (2.3 ±0.6) mg· L-1 forcmax, (48 ±14)mg · h · L-1 and (48 +14) mg· h · L-1 forAUC0-60,(51+15) mg· h· L-1 and (50±14) mg· h· L-1for AUC0-∞, (22.4 ± 1.9 ) h and (22.8 ± 1.9 ) h for MRT, respectively. The steady state pharmacokinetic parameters after multiple doses of the domestic and imported gliclazide tablets were (6. 1 ± 1.4) h and (6.5+1.4) h for tmax, (4.6±0.9) mg· L-1 and (4.7±1.1) mg· L-1 for cmax, (0.23 ±0.08) mg ·L-1and (0.26±0.08) mg· L-1 forcmin, (1.6±0.3) mg·L-1 and (1.6±0.3) mg · L-1 for mean value of steady plasma-drug concentration (cav),(94±19) mg· h · L-1 and (95 ±20) mg · h · L-1forAUCss, (282 ±33)% and (283 ±43)% for degree of fluctuation DF ), respectively. The relative bioavailability of the domestic gliclazide tablet to the imported gliclazide tablet following a single and multiple dose were ( 102 ± 9) % and (99 ± 10 ) %, respectively. Main pharmacokinetic parameters between the two formulations in both single and multiples dose studies showed no statistical difference ( P >0.05 ). CONCLUSION: The result of two one side t-test shows that the two formulations are bioequivalent.
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Objective:To find out markers of endometriosis. Methods: The two dimensional gel images of proteins extracted from eutopic endometrium from endometriosis patients and controls were analyzed by software Phoretix 2D,and the proteins expressed differently were identified primarily by query of data base.The proteins extracted from ectopic endometrium of ovarian endometriosis were transferred from two dimensional gel onto nitrocellulose membranes,followed by incubation with sera from women with and without endometriosis. Analyzed by MALDI-TOF-MS,the proteins hybridized differently were identified through their Peptide Mass Footprints. Results: Having compared the reproducible two dimensional gel images of proteins from eutopic endometrium of women with and without endometriosis,we obtained 11 proteins expressed differently.Through Western Blot technique,we found three proteins hybridized differently which were identified as vimentin,?-actin and ATP synthase ? subunit respectively. Conclusion: The protein expression spectra of eutopic endometrium from patients with endometriosis are significantly different from those of the controls,and the anti-endometrial autoantibodies against vimentin, ?-actin and ATP synthase ? subunit may be induced.
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Objective: To screen out serum protein profiling of polycystic ovary syndrome(PCOS) by surface - enhancedlaser desorption/ ionization time of flight mass spectrometry (SELDI-TOF MS) for discovering the discriminatory proteins. Methods: Thirty one women with PCOS and thirty healthy women were detected by Weak Cation Exchange(WCX2)chip. ProteinChip reader and Biomarker Wizard software from Ciphergen Inc were combined with a bioinformatics method (support vector machines, SVM ) to analyze protein fingerprinting. Results: There were 4 proteins which were obviously different between the PCOS group and the control. Three of them were up-regulated, and one down-regulated. To set up a analysis model by SVM using the 4 proteins could successfully distinguish between PCOS and the normal control. The corresponding sensitivity, specificity and positive predict value were 86.7%, 83.3%, 87.2%, resectively. Conclusion: Using ProteinChip technology can screen out serum discriminatory proteins quickly and efficiently. Combined with SVM, an optimal fingerprinting model has been set up, which can easily predict PCOS. In disease state of PCOS, there are significant variations which consist of four proteins. To investigate those four discriminatory proteins, especially the protein m/z 6 628 may be of benefit to pathogenic study and the development of biomarkers for PCOS.
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As a crucial technique for proteome analysis, mass spectrometry (MS) has developed rapidly in the last two decades. MS has established itself as a powerful tool in life science by its high sensitivity, resolution, mass accuracy and the ability to be coupled with chromatogram and isotope labeling. This review discusses the principles and instrumentation developments of MS, especially focuses on the applications of MS in proteomic studies.
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Objective:To establish a high performance liquid chromatography/tandem mass spectrometry(HPLC-MS) method for determination of cyclosporin A level in the whole blood.Methods: A Hypersil C_(8) column(65 ℃,4.6 mm ?150 mm,5 ?m) was used with a mobile phase of acetonitrile-acetic acid(pH 4.50) 7030 at a flow rate of 1.0 ml?min~(-1).Atomospheric pressure electronic spray ionization was used to quantify cyclosporin A,with the capillary voltage being 4 000 V,the flow of drying gas being 8.0 L?min~(-1),nebulizer pressure being 30 psi,and fragment voltage being 140 V.Results: The calibration curve of cyclosporin A was linear in a range of 26.68-1 067 ?g?L~(-1): y=0.001 1x-0.004 6(r=0.999 3).The lowest limit of detection was 4.35 ?g?L~(-1).The method recoveries of low,medium and high concentrations cyclosporin A were((89.81?4.91)%,)(104.5?3.73)%,and(95.17?2.33)%,respectively;the extraction recoveries of 3 concentrations were(77.64?4.38)%,(78.01?3.79)%,and(83.02?1.59)%,respectively.The intra-day and inter-day RSD was less than 6%,and the RSD was less than 5% in 150 days.Conclusion: The present method is fast,sensitive and selective.It is suitable for clinical monitoring of cycolsporin A.
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Objective:To analyze the chemical constituents of the ether extracts from the fruits of 3 species of Chaenomeles . Methods: The chemical constituents were analyzed by GC MS. Results: Sixty one constituents were firstly isolated and identified from the fruits of 3 species of Chaenomeles . Forty compounds in Chaenomeles lagenaria , thirty four compounds in Xuan mugua ( C.lagenaria from Xuanzhou) and 23 compounds in C.sinensis were isolated and identified respectively. There were 10 compounds in all 3 species of Chaenomeles , but their contents were very much different. They were acetic acid, benzaldehyde, n caproic acid, 14 methyl pentadecanoic methyl ester, glycerin, benzoic acid, 6,9 octadecadienoic methyl ester, squalane, stearic acid and oleic eicosyl ester. Conclusion: The composition and content vary with different species and district of Chaenomeles . The result may be useful for the evaluation of Chaenomeles fruits quality. [
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Se realiza el estudio mediante técnicas cromatográficas, de un grupo de cepas de Mycobacterium tuberculosis aisladas de un brote en pacientes infectados con el virus VIH. Se utilizaron como referencia un grupo de cepas de M. tuberculosis de pacientes sintomáticos respiratorios (SR+14) y cepas patrones de la colección del laboratorio, con el objetivo de comparar cualitativamente los patrones cromatográficos descritos por las cepas aisladas de pacientes. Se obtuvieron y compararon los perfiles cromatográficos de las cepas aisladas de pacientes (ST+) y de VIH+ por la técnica de cromatografía en capa delgada. Se identificó cada uno de los ácidos grasos presentes por la técnica de cromatografía de gases acoplada a espectrometría de masas. Todas las cepas estudiadas fueron clasificadas como Mycobacterium tuberculosis. Por resultados obtenidos se demuestra la utilidad de las técnicas cromatográficas como técnicas alternativas para el diagnóstico micobacteriano.
A group of strains of Mycobacterium tuberculosis isolated from an outbreak in HIV-infected patients was studied by chromatographic techniques.A group of strains of M. Tuberculosis from symptomatic respiratory patients (SR + 14) and patterns strains from the laboratory collection were used as a reference aimed at making a qualitative comparison of the chromatographic patterns described by the strains isolated from patients. The chromatographic profiles of the strains isolated from patients (SR +) and fro HIV + were obtained and comparede by thin layer chromatography (TLC). Each of the present fatty acids was identified by using the gas chromatography technique (GC) coupled to mass spectrum analysis. All the studied strains were classified as Mycobacterium tuberculosis. According to the results attained, the usefulness of the chromatographic techniques as alternative techniques for the mycobacterial diagnosis is demonstrated.
Subject(s)
Humans , AIDS-Related Opportunistic Infections/microbiology , Disease Outbreaks , HIV-1 , Mycobacterium tuberculosis/chemistry , Tuberculosis, Pulmonary/microbiology , Cuba , Gas Chromatography-Mass Spectrometry/methods , Gas Chromatography-Mass Spectrometry/statistics & numerical data , Mycobacterium tuberculosis/isolation & purification , Reference StandardsABSTRACT
Objective To establish a simple, sensitive and accurate analytical method to determine 15 elements in drinking water-related products soak test. Methods Inductively coupled plasma-mass spectrometry (ICP-MS) was employed to determine 15 elements in the soaking liquid. Sc, Ge, In and Bi were used as the internal standard substances. Results The test results showed the linear correlation coefficients for all elements were more than 0.999, the recovery rate were 80.6%-120.0% and precision was fine, RSD was lower than 6.0 %, the test results also showed a close agreement with the reference values in test of reference materials of GSBZ 50009-88, GSBZ 500019-90, GSBZ 50004-88 and GSBZ50039-95. Conclusion This method is simple, rapid, sensitive, accurate and suitable for determination of 15 elements in drinking water-related products soak test.
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In recent years,great progress has been made in the theory and technology of proteomics.As embranchment of proteomics,serum proteomics has been payed more and more attention also.The advanced techniques have been applied in serum proteomics research,which promote the development of the methodology.A great advance has been made in serum proteomics research for seeking associated tumor markers,pharmacy and some non-cancer diseases.