Improved Mass Determination of Poly(ethylene glycols) by Electrospray Ion-Mobility Time-of-Flight Mass Spectrometry Coupled with Ion–Molecule Reactions - Pharmaceutical Technology

Latest Issue
PharmTech

Latest Issue
PharmTech Europe

Improved Mass Determination of Poly(ethylene glycols) by Electrospray Ion-Mobility Time-of-Flight Mass Spectrometry Coupled with Ion–Molecule Reactions
The authors developed a method to accurately measure the average molecular weight of large poly(ethylene glycols) (PEGs) using ion-mobility time-of-flight mass spectrometry coupled with gas-phase ion–molecule reactions.


Pharmaceutical Technology


Experiment

Chemicals and reagents. Methanol (CH3OH) and isopropanol (IPA) were purchased from Fisher Scientific (Pittsburgh, PA). Ultrapure water (18.2 M) was obtained from a Milli-Q water purifier (Millipore Corporation, Medford, MA). Ammonium acetate, cesium iodide and superbase, 1,8-diazabicyclo[5.4.0]-undec-7-ene (DBU), were purchased from Sigma (St. Louis, MO). PEG 4450 was obtained from Waters Corporation (Milford, MA). PEG-aldehyde 20K was purchased from NOF Corporation (Tokyo, Japan). All materials were used without additional purification.

Sample preparation. All polymer samples were prepared at analyte concentrations of 1–2 mg/mL (10-4 M based on the nominal molecular mass) either in a 50/50 (v/v) solution of water and methanol or 10 mM ammonium acetate and methanol. The solutions were directly infused into the ESI source at 10 L/min by a syringe pump (Harvard Apparatus, Holliston, MA).

Mass spectrometry. All experiments were performed on a Synapt high-definition mass spectrometer, a hybrid quadrupole ion-mobility TOF mass spectrometer (Waters Corp.). Detailed descriptions of the instrument was found elsewhere (17, 18). A schematic diagram of the instrument is shown in Figure 1a.


Figure 1a & 1b
The instrument was operated in positive-ion mode with a capillary voltage of 3.0 kV. The sampling cone voltage was set at 40 V. The ion source block and nitrogen desolvation gas temperatures were set to 100 C and 250 C, respectively. Nitrogen was used as both the nebulizing gas and the desolvation gas. The desolvation gas was set to a flow rate of 250 L/h. The quadrupole was operated in nonresolving mode to transmit a wide m/z range.

The instrument can be operated in two modes: TOF mode and TOF-mobility mode. For TOF-mobility experiments, the ion mobility cell pressure was maintained at 0.5 mbar (N2) and separations were optimized by tuning the wave pulse height from 5 to 14 V. The transfer cell pressure was maintained at 0.02 mbar unless stated otherwise. Pressures in the trap and IMS cell were the direct readbacks from the active pirani gauges equipped with the instrument. The RF amplitude used on the trap, ion mobility cell, and transfer cell was 320 V peak-to-peak. Ion mobility cell traveling wave velocities of 240 m/s were used.

The instrument was externally mass calibrated with cesium iodide (1.0 mg/mL in 50% IPA–50% water). Data acquisition and processing were controlled by MassLynx 4.1 software (Waters Corp). Mass spectra were acquired every 1 s in TOF mode and every 2 s in TOF-mobility mode from m/z 100 to m/z 6000 for PEG 4450 samples and from m/z 500 to m/z 10,000 for PEG-aldehyde 20K for 6 min. Additional data processing to yield the information on the average molecular weight and the polydispersity of the PEGs followed the methods described below.


ADVERTISEMENT

blog comments powered by Disqus
LCGC E-mail Newsletters

Subscribe: Click to learn more about the newsletter
| Weekly
| Monthly
|Monthly
| Weekly

Survey
FDASIA was signed into law two years ago. Where has the most progress been made in implementation?
Reducing drug shortages
Breakthrough designations
Protecting the supply chain
Expedited reviews of drug submissions
More stakeholder involvement
Reducing drug shortages
29%
Breakthrough designations
10%
Protecting the supply chain
43%
Expedited reviews of drug submissions
10%
More stakeholder involvement
10%
View Results
Jim Miller Outsourcing Outlook Jim Miller Health Systems Raise the Bar on Reimbursing New Drugs
Cynthia Challener, PhD Ingredients Insider Cynthia ChallenerThe Mainstreaming of Continuous Flow API Synthesis
Jill Wechsler Regulatory Watch Jill Wechsler Industry Seeks Clearer Standards for Track and Trace
Siegfried Schmitt Ask the Expert Siegfried SchmittData Integrity
Sandoz Wins Biosimilar Filing Race
NIH Translational Research Partnership Yields Promising Therapy
Clusters set to benefit from improved funding climate but IP rights are even more critical
Supplier Audit Program Marks Progress
FDA, Drug Companies Struggle with Compassionate Use Requests
Source: Pharmaceutical Technology,
Click here