6410B Triple Quadrupole LC/MS from Agilent Technologies

May 20, 2013

Overall

Performance

Ease-of-Optimization

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Mercer University, Pharmacy Sciences
Pharmaceutical
PhD Pharmaceutics
Shown in the figure is a standard seven point calibration curve showing relative peak abundance response ratio using sodium lauryl sulfate d-25 as an internal standard from an extracted batch of sodium lauryl sulfate from a topical ointment formulation demonstrating excellent linearity from 250-2000 ng/mg.

Company:

Agilent Technologies

Product Name:

Agilent 6410B Triple Quadrupole LC/MS

Catalog Number:

6410B
Image
  • Company: Agilent Technologies
  • Product Name: Agilent 6410B Triple Quadrupole LC/MS
  • Catalog Number: 6410B

Much of our current research involves drug formulation for optimized drug delivery systems in the pharmaceutical sciences. With this goal in mind, accurate quantification for evaluation of active pharmaceutical agents in our delivery vesicles is essential. As a gold standard in modern pharmaceutical analysis, liquid chromatography tandem mass spectrometry plays a large role in the analysis of known and unknown compounds, polar and nonpolar alike, and confers selective advantages such as high sensitivity and selectivity for the target analyte. The Agilent 6410B LC/MS is a triple quadruple mass spectrometer.

Experimental Design and Results Summary

Application

Single and multiple reaction monitoring

Starting Material

Sample extracts, biologics, peptides. A specific example of a drug we assayed was an inactive excipient Sodium Lauryl Sulfate present in topical lotions and ointment as a surface active agent or surfactant.

Protocol Overview

The Agilent 6410B LC/MS is a triple quadruple instrument where analytes are introduced into the ion source from a liquid eluent and are further converted into gaseous state molecules by the process of desorption. These molecules are then transferred through a vacuum into an ion transfer capillary and passed through the first quadrupole for mass filtering. From the first quadrupole, a precursor ion may be selected; this ion is further allowed to pass into a collision cell where collision induced dissociation (CID) occurs and the precursor ion is fragmented into various product ions. The third quadrupole acts as the secondary mass filter for selection of target ions after CID. The overall monitoring of a selected transition from precursor to product ion is known as single reaction monitoring, when multiple ions are monitored the term multiple reaction monitoring is used.

Tips

For method development, a nice new feature is the Masshunter optimizer where the analyte can be infused directly into the ionization source or by a series of injections, the software can pickup on selected product ions as well as optimize fragmentor voltage and collision energies from a mix of analytes assayed at once, you can simply walk away, come back and have all the parameters available.

Results Summary

See graph below

Features Summary

The instrument has a nice core software package, Masshunter, which is very easy to use with a user-friendly interface. When starting the data acquisition software, four panes are immediately loaded: instrument status pane, a real time acquisition pane showing a total ion chromatogram and the mass spectra, a method pane for LC and MS instrumental parameters, followed by a worklist pane detailing the sample set. With Agilent’s user-friendly software, a new run can be setup in a matter of minutes. To run a sample, data acquisition is loaded whereby a method is then loaded, a sample set is entered into a new worklist, and the injections begin. For data analysis and interpretation, two software packages are included with the instrument: Masshunter qualitative analysis and Masshunter quantitative analysis. For real-time monitoring of data acquisition, including TIC and mass spectra, data acquisition software is used to monitor up to 60 minutes of real-time data. The core package used for assessment of sample worklists and calibration is quantitative analysis. The software works by first selecting a new batch and by adding samples, a quantitative method is then built with the series of standards and an appropriate internal standard is selected, the method is saved and the batch is analyzed. Agilent provides a series of peak integration scripts for the calibration analysis. The software is very easy to use and can be learned quickly for new users. Another nice thing about the instrument compared to other instruments is how easily the system tunes up. Before running analytical assays, we routinely run a check-tune and periodically run an auto-tune to ensure that the mass spectrometer is functioning correctly. For instruments from other vendors, it can sometimes be troublesome to meet the criteria from daily standard operating procedure requirements before running sample batches. In the case of the Agilent 6410B, only minor problems occurred and 95% of the time the instrument tunes up to spec. Another nice feature is the autotune polymer mix for QQQ and QTOFs, the solution does not have to be prepared fresh or diluted, with no manual infusion involved. There is a storage region for the solution and the instrument will auto inject the tune mix under tune mode from data acquisition. Tuning the mass spec could be quite time consuming with periodical challenges, but Agilent simplifies the procedure with ease.

Additional Notes

Over a period of a week, a very accommodating Agilent service engineer specialist brought the instrument to our department for installation and evaluation. We tested the sensitivity of the instrument immediately using a series of Reserpine injections.

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Shown in the figure is a standard seven point calibration curve showing relative peak abundance response ratio using sodium lauryl sulfate d-25 as an internal standard from an extracted batch of sodium lauryl sulfate from a topical ointment formulation demonstrating excellent linearity from 250-2000 ng/mg.

Summary

The Good

Rapid, accurate, precise, sensitive, selective, high reproducibility, multiple ionization sources available.

The Bad

Expensive to maintain, gas consumption, need a dedicated user, repairs may be very costly.

The Bottom Line

The instrument is overall very well designed. If looking for an instrument to run any sort of pharmaceutical bio-analytical work, try out the 6410B QQQ; your laboratory will not be disappointed.

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