Application Note 01546

Residual Solvent Analysis Using the Teledyne Tekmar HT3 Headspace Autosampler and Varian 4000-MS GC/MS Ion Trap Mass Spectrometer
Ron Honnold, adapted by Tiffany Payne from a Tekmar application note1 Varian, Inc.

Introduction
Many solvents are used in the manufacturing of pharmaceutical products, substances and excipients. In order to protect the health of the patient, pharmaceutical companies have adopted methods that will reduce the toxicity of the residual solvents. The International Conference on Harmonization (ICH) of Technical Requirements for Registration of Pharmaceuticals for Human Use has published guidelines on a list of three separate classes of residual solvents and their daily exposure limits.2 Class 1. These residual solvents are known carcinogens or environmental hazards and are avoided whenever possible in pharmaceutical manufacturing. Class 2. This class causes some reversible or irreversible toxicity but is less toxic than Class 1. These solvents are limited in their use in pharmaceutical manufacturing. Class 3. These residual solvents have a low toxic potential or have no health related exposure limits. The Varian 4000 GC/MS Quadrupole Ion Trap provides sensitive and accurate analysis for residual solvents.

GC/MS Conditions
Injector Temp: 280 oC Split State: 100:1 Carrier Gas: Helium at 1.5 mL/min, EFC, constant flow mode Column Temp: 40 C, 3.0 min hold, 5.0 C/min to 60 C, 8.0 C/min to 140 C, 30 C/min to 280 C, 1.0 min hold Total Run Time: 23.4 min Column Type: FactorFour 60 m x 0.25 mm fused silica column coated with 0.25 micron VF-5ms (Varian part no. CP4989) Trap Temp: 220 oC Manifold Temp: 45 oC Transfer Line Temp: 310 oC Filaments: 10 A Multiplier: -200 V Target: 8000 counts Max Ion Time: 15000 sec Scan Speed: Fast

Instrumentation
Varian 4000 GC/MS Quadrupole Ion Trap using internal ionization mode Varian CP-1177 Split/Splitless (S/SL) Injector Teledyne Tekmar HT3 headspace autosampler

Results & Discussion

Table 1 provides calibration statistics for a calibration from 50 to 1000 ppm.
Table 1. Calibration curve from 50-1000 ppm.
Compound Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Compounds Methyl Alcohol Ethanol Acetonitrile Acetone Isopropyl Alcohol Methylene Chloride 1-Propanol 2-Butanone Ethyl Acetate Trichloromethane Tetrahydrofuran 1,2-Dichloroethane 1-Butanol Benzene Cyclohexane Trichloroethylene 1,4-Dioxane MIK Toluene N,N-Dimethylformamide Chlorobenzene N,N-Dimethylacetamide DMSO %RSD 20.34 12.10 13.42 4.74 8.79 7.96 13.80 7.58 5.56 9.24 5.77 6.69 10.59 8.62 6.21 5.25 5.82 7.72 5.93 6.37 5.05 13.71 (r2) 0.9979 0.9988 0.9986 0.9993 0.9993 0.9991 0.9972 0.9987 0.9992 0.9993 0.9993 0.9988 0.9971 0.9992 0.9973 0.9993 0.9993 0.9991 0.9994 0.9985 0.9989 0.9967

Sample Preparation
To extract and analyze the residual solvents from the drug product, the drug product itself must first be dissolved. For this method dimethyl sulfoxide (DMSO) was used as the solvent. Once the drug compound is dissolved in a headspace vial, the vial itself is placed on the HT3 headspace autosampler. The static headspace sample introduction technique allows residual solvents in DMSO to diffuse into a gas phase within a sealed vial until equilibrium is reached. The gas phase is then transferred to the GC/MS for analysis. There is minimal interference from the drug substance because only the gas phase is transferred to the analytical system. A working standard of 2500 ppm residual solvents was prepared in DMSO and dilutions were made from that working standard also using DMSO. A five-point calibration curve was analyzed using 50, 100, 250, 500 and 1000 ppm standards (5 mL of each standard was added to a 22-mL headspace vial). The loop volume used on the HT3 (injection volume into the GC/MS) was 0.25 mL.
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Repeatability studies showed that for six samples at 500 ppm, the %RSD for the target analytes ranged from 3.3% (isopropyl alcohol) to 4.8% (trichloromethane). In addition, MDL studies showed that for seven samples at 50 ppm the %RSD ranged from 0.03% (methyl alcohol and tetrahydrofuran) to 0.06% (acetonitrile and acetone). The MDL values ranged from 5.46 ppm (tetrahydrofuran) to 10.24 ppm (acetonitrile).

Figure 3. Blank injection chromatogram after a 1000 ppm injection. *Represents dimethylsufone, not one of the target analytes for this analysis.

Conclusion
The Tekmar HT3 headspace autosampler with the Varian 4000 GC/MS Ion Trap Mass Spectrometer provides a sensitive, robust and quantitative analytical system for the analysis of residual solvents in pharmaceutical products. Headspace provides a cleaner alternative to methods that involve direct injections. A cleaner analysis means less GC maintenance required, which ultimately reduces downtime and increases productivity.

Figure 1. Sample chromatogram at 50 ppm. Numbers correspond to compound numbers in Table 1. *Denotes dimethylsufone, not a target analyte.

References
1. Residual Solvents by Teledyne Tekmar HT3 Headspace Autosampler and Varian 4000 GCMS. An application note by Tekmar, a Teledyne Technologies company. February 15, 2007. 2. International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (Impurities: Guideline for Residual Solvents). Recommended for Adoption at Step 4 of the ICH Process on 17 July 1997 by the ICH Steering Committee.
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2008 Varian, Inc.

Figure 2. Sample chromatogram at 1000 ppm. Numbers correspond to compound numbers in Table 1. *Denotes dimethylsufone, not a target analyte.

These data represent typical results. For further information, contact your local Varian Sales Office. Varian, Inc. www.varianinc.com North America: 800.926.3000 925.939.2400 Europe The Netherlands: 31.118.67.1000 Asia Pacific Australia: 613.9560.7133 Latin America Brazil: 55.11.3238.0400
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