Category: Formulation and Quality
Purpose: Microcrystalline cellulose is used as an excipient in the pharmaceutical formulations. Excipients play a major role in the performance and stability of the drug product. Many review articles reveal that the presence of trace level of reactive impurities like formaldehyde and acetaldehydes in pharmaceutical excipients may cause drug product instability leading to the formation of degradants, or loss in the drug product performance. Hence, the determination of the presence of trace levels of formaldehyde and acetaldehyde in microcrystalline cellulose is of importance. There are few literature exampless available for the determination of formaldehyde and acetaldehyde content by HPLC and GC method. The objective of this study was to develop and validate an HPLC method to measure the formaldehyde and acetaldehyde content in microcrystalline cellulose.
Methods: The present developed method is based on the reaction of formaldehyde and acetaldehyde with 2,4-dinitrophenylhydrazine (2,4-DNPH). The method involved weighing of 0.2 grams of microcrystalline cellulose sample into centrifuge tubes and addition of 5 mL of 2,4-DNPH solution, vortexed for 2 minutes. The sample tubes are placed into a water bath maintained at 60 °C for one hour with intermittent shaking at every 15 minutes. After one hour, the sample tubes are removed, vortexed and allowed to cool to room temperature. The samples are centrifuged, and a clear supernatant solution is transferred to an HPLC vial for analysis. The samples are analyzed by HPLC method using mobile phase of water, methanol, acetonitrile and flush phase consisting of acetonitrile and acetone with gradient program and C18 HPLC column at wavelength of 360 nm. The quantification was done by an external standard method based on peak area.
Results: The method has been successfully developed and validated for determination of formaldehyde and acetaldehyde content in microcrystalline cellulose. The method has been validated for accuracy, precision, linearity, limit of detection and limit of quantification. The method was found to be linear in the range of 0.35 ppm to 150.67 ppm with correlation coefficient of 0.999 for formaldehyde and 0.03 ppm to 145.72 ppm with correlation coefficient of 0.999 for acetaldehyde, respectively.
Conclusion: A sensitive and accurate method has been developed and validated for determination of formaldehyde and acetaldehyde content using 2,4-dinitrophenylhydrazine as derivatizing agent. This method has been successfully applied for the formaldehyde and acetaldehyde content estimation in microcrystalline cellulose samples.
Kevin O'Donnell– R&D Manager, DuPont Pharma Solutions, Midland, Michigan
Rathnakar Palarapu– Hyderabad, Telangana, India
Sumakala Sibyala– Hyderabad, Telangana, India
Vinay Muley– Team Leader, Dow Dupont, Hyderabad, Telangana, India
Markus Mintert– Bomlitz, Hamburg, Germany
Barbara Serr– Midland, Michigan