Sciex API

Perkin-Elmer Sciex API 3000 LC/MS/MS system Perkin-Elmer Sciex Turbo lonSpray electrospray interface... 

Using the Tomtec Quadra 96 workstation, 0.1 mL of the ethyl acetate layer was transferred to a 96-well collection plate containing 0.4 mL of acetonitrile in each sample well. The solution was mixed 10 times by aspiration and dispersion on the Tomtec. The plate was then covered with a sealing mat and stored at 2 to 8°C until LC/MS/MS analysis. The HILIC-MS/MS system consisted of a Shimadzu 10ADVP HPLC system and Perkin Elmer Sciex API 3000 and 4000 tandem mass spectrometers operating in the positive ESI mode. The analytical column was Betasil silica (5 fim, 50 x 3 mm) and a mobile phase of acetonitrile water formic acid with a linear gradient elution from 95 5 0.1 to 73.5 26.5 0.1 was used for 2 min. The flow rate was 1.0 mL/min for the API 3000 and 1.5 mL/min for the API 4000 without any eluent split. The injection volume was 10 jjL and a run time of 2.75 min was employed. 

A six-port valve was used in both manual and semi-automated SPME interfaces and PEEK tubing used to connect the HPLC system to the SPME probe. A Cohesive HTLC 2300 with dual pumps along with a Sciex API 3000 mass spectrometer was used for LC/MS/MS and a Symmetry Shield RP-18 (5 ji, 50 x 2.1 mm) for HPLC. A quaternary pump with flow switching was used for desorption chamber flushing along with MS make-up flow and a binary pump for LC/MS/MS. Acetoni-trile/0.1% acetic acid in water (90 10, solvent B) and 10 90 acetonitrile/0.1% aqueous acetic acid (solvent A) were used, with 10% B for 0.5 min ramped to 90% B in 2 min and held at this concentration for 1.5 min before returning to 10% B for 1 min at a flow rate of 0.5 mL/min. 

Recently, Chu et al. reported an ultra-fast LC/MS method for analysis of cytochrome P450 3A4 and 2D6 inhibition assaysd Testosterone and dextromethorphan were used as the specific substrates for CYP3A4 and CYP 2D6, respectively. LC/MS analyses were performed on a Sciex API 3000 mass spectrometer equipped with a Shimadzu LC-lOAdvp pump and a PE 200 autosampler. A Phenomenex Luna CIS (4.6x30 mm) column was used along with very steep gradients. Each sample analysis was completed in 0.5 min. 

SCIEX API 365 equipped with a TurboIonSpray interface operated in positive ion mode. The calibration range 0.124-497 ng/mL was readily validated with a negligible carry-over effect from this system. The method offered a total cycle time of 8 min and completely eliminated the manual sample preparation. ... 

Mass spectra were recorded on a mass spectrometer (PE Sciex API III triple-quadrupole) interfaced with a Sciex Ion-Spray probe. Liquid chromatography was performed with a pump (Perkin Elmer Binary 250) and a PDA Detector (LC480 Auto Scan). The separation was achieved using a standard linear gradient (80% 2 mM ammonium acetate solution,... 

Sciex API 5000, triple quadrupole Turbo lonSpray/positive... 

Figure 1.14. MRM chromatograms of SCH 29851 (383.0. 337.0) and SCH 34117 (311.1 259.1) obtained using Sciex API 3000 (triple-stage quadrupole) and Sciex QSTAR pulsar (Q-TOF). Comparison of MRM chromatograms of SCH 29851 and SCH 34117 obtained at the LOQ (1 ng/mL) using the API 3000 mass spectrometer with those from the Q-TOF mass spectrometer indicated that the S/N ratio is at least 10-20 times better on the API 3000 mass spectrometer. However, the MRM chromatograms from the API 3000 mass spectrometer do not provide the option to further examine the MS/MS spectra whereas the full-scan MS/MS spectra from a Q-TOF based quantitative bioanalysis assay allows one to easily eliminate any questions about false-positive data. (Rephnted with permission from Yang et a ., 2001b.)...

An atmospheric pressure chemical ionization (APCI-LC-MS) (Sciex API 150 EX) method was developed for the determination of zaleplon and zolpidem in the whole blood. After single-step LLE, the hypnotics were separated by gradient-elution with an ammonium formate buffer/acetonitrile eluent on an Inertsil ODS-3 column. Methaqualone was used as IS. The recovery was higher than 70% for both hypnotics and the IS. The method was successfully applied to forensic cases [11]. 

A PE-Sciex API-I lonSpray mass spectrometer (PE-Sciex, Thornhill, Ontario, Canada) was used to acquire all mass spectra. The Sciex API-Ill mass spectrometer and Integral micro analytical workstation were coupled through a Im piece of fused silica tubing (75 im i.d.) at the exit of the capillary flow cell detector using a 250pm i.d. teflon sleeve as described previously (8). The source... 

N-terminal sequence analyses were performed using ABI 477/120A or 473A protein sequencers. For mass spectrometric analyses, aliquots of the collected peak fractions were also concentrated in a Savant SpeedVac to 20 pmol/pL and infused using a Harvard syringe pump into a PE SCIEX API III electrospray mass spectrometer (ESI-MS) operating in the positive mode with an orifice potential of 80 volts. 

Electrospray mass spectra were recorded on a Perkin-Elmer Sciex API-Ill triple quadrupole mass spectrometer (Thornhill, Canada) with general MS and LC setup as previously described (5-8). Modifications of the instrument which improve sensitivity for negative-ion mode detection of phosphorylation and sulfation have also been described before (7,8). Because phosphorylated or sulfated peptides produce predominantly negatively charged marker ions while glycosylation or acylation yield positively charged species, these two classes of modification cannot both be detected in a single LCMS experiment. 

Protein Sequencing Mass Spectrometry Proteins were sequenced on an Applied Biosystems 477A protein sequencer. Polypeptide molecular weights were determined after analysis by a Sciex API-Ill electrospray ioniziation mass spectrometer. 

All nES/MS and nES/MS/MS measurements were made on a Perkin-Elmer Sciex API-in triple quadrapole mass spectrometer (Thornhill, Canada) equipped with the nanoelectrospray source designed by Matthias Wilm and Matthias Mann at the European Molecular Biology Laboratory, Germany (9). The long signal duration allowed the instrumental parameters to be optimi for each peptide interrogated by nES/MS/MS. Argon gas was used as the collision... 

The solution of [15N]r-metHuLeptin (0.3 mg/ml) in PBS (0.1 M sodium phosphate, 0.1 M sodium chloride, pH 7.2) was incubated with endoproteinase Asp-N at an enzyme-to-substrate ratio of 1 75 (w/w) at 25 °C for 5 h. The digestion was terminated by adding 5 )tL of 5% TFA to the reaction. Peptides were separated by a Vydac C4 reverse-phase analytical column (4.6x250 mm) using a Hewlett Packard HPLC (Model 1090), which is on-line connected to a PE-Sciex API-100 electrospray mass spectrometer. The column was initially equilibrated with 95% mobile phase A (0.1%TFA) and 5% mobile phase B (90% acetonitrile in 0.1% TFA). A linear gradient from 10 to 50% mobUe phase B was run over a period of 85 minutes at a flow rate of 0.5 raL/min. The splitting of the flow (9 1) was achieved post UV cell, allowing 50 iL/min of the eluent to be analyzed by the electrospray mass spectrometer. 

Observed mass on Sciex API 100 Plus with ES source. 

The molecular mass of protein and peptide samples was determined by electrospray ionization mass spectrometry using a Perkin-Elmer Sciex API 100 mass spectrometer. The sample was introduced either by infusion or by on-line liquid chromatography/mass spectrometry (LC/MS) using a splitter. The data were obtained by scanning from 450 to 2000 Da with a scan time of 5 s and a step size of 0.25 Da with 1.0 ms dwell time per mass step. The molecular mass of the sample was obtained using the software provided by the instrument manufacturer. 

Mass Spectrometry and Other Analytical Procedures. Liquid chromatography electrospray mass spectrometry (LC-ESMS) was performed on approximately 1 pg protein samples with a Perkin Elmer Sciex API-300 triple quadrupole mass spectrometer fitted with an articulated ion spray source and set to scan over a range of 400-3000... 

Thermodynamics of Enzyme and Inhibitor Interactions SCIEX API III mass spectrometer. 

Electrospray mass spectra were recorded in a PE Sciex API-111 mass spectrometer as described previously (10, 16). A capillary HPLC was connected on-line to the electrospray interface of the mass spectrometer. 

Liquid chromatography-mass spectrometry (LC-ESI-MS) was performed on a PE-Sciex API-Ill triple quadrupole mass spectrometer with an lonSpray source. The samples were separated on the Nucleosil Cl 8 column with gradient 1 a splitting device was used to direct a portion of the effluent into the mass spectrometer. Data were acquired in the range of m/z 400-1000, with sufficient resolution to detect the isotope peaks for singly-charged ions at m/z 1000. 

By collecting residues from the curtain plate in a Sciex API source (Figure 5.6) and re-injection, it was demonstrated that nonvolatiles present in the liquid flow, originating from nonvolatile mobile-phase additives or sample constituents actually increase the amount of analyte found at the curtain plate. The nonvolatiles prevent the preformed analyte ions to desorb from the droplet into the gas phase. This study emphasizes the importance of removing nonvolatile sample constituents during sample pretreatment [76]. It was concluded that APCI is less prone to matrix effects than ESI, which is also confirmed by others, e.g., [77-79], but one should certainly not exclude the occurrence of matrix effects in APCI-MS. 

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