"Investigating UPC2 Method Development Strategies and Applying them to Impurity Profiling of Pharmaceutical Products"
Michael D. Jones
Waters Corporation


Converging Chromatographic Approaches:  A Critical Evaluation of Sub-2µm particles and Supercritical CO2 mobile phases
Michael D. Jones
Waters Corporation



Schedule:

"Investigating UPC2 Method Development Strategies and Applying them to Impurity Profiling of Pharmaceutical Products"
Michael D. Jones
Waters Corporation


Abstract
Innovator and generic pharmaceutical companies invest much time and resources in R&D focused on impurity analysis to fulfill regulatory requirements for NDA and ANDA submissions.  The ICH guidelines outline thresholds for impurities in pharmaceutical drug substances and drug products for reporting, identification and qualification.  Analytical techniques for the quantification of impurities must be capable of detection levels as low as 0.05% relative to the active pharmaceutical ingredient.  In addition, pharmaceutical companies often demonstrate drug safety by using orthogonal analytical techniques to assure the separation of all substances.  For generic pharmaceutical companies, the impurity profiles of their drug are compared to the innovator's pharmaceutical product by means of orthogonal analytical techniques to prove equivalence.  Integrating detection methods such as photodiode array and mass spectrometry provide the spectral documentation necessary for the industry to demonstrate the quality of the drug substance and drug product.  Recently, a new separation technique has emerged addressing this often challenging application.

UltraPerformance Convergence Chromatography (UPC2) exploits the benefits of a sub-2µm particle size stationary phase, with carbon dioxide as the primary mobile phase component.  The use of convergence chromatography is an attractive alternative to liquid chromatography that provides different selectivity, low solvent use, high efficiencies and aqueous-free environments.  However, while method development strategies in LC and GC chromatography are well defined, that is not the case for convergence chromatography.  In order to streamline this process, a systematic approach to achiral convergence chromatography method development requires research. 

In this presentation, columns and solvents were screened to determine suitable method conditions providing separation of the analytes yielding optimal peak shape.  We will evaluate the chromatographic behavior and correlate the results to the chemical properties of the analyte of interest.  The proof of concept approach will be used to characterize optimal operating conditions when implementing ACQUITY UPC2 method development activities.  

As an example, ACQUITY UPC2 was utilized to analyze Metoclopramide and related impurities at limits specified by ICH Q3A and Q3B guidelines.  Metoclopramide is an antiemetic drug used for the treatment of heartburn, healing of ulcers, and nausea when undergoing chemotherapy.  Various methods of analysis for metoclopramide use reversed-phase and hydrophilic interaction chromatographic techniques depending on the dosage form .2-3    The European Pharmacoepoeia indicates six known impurities with additional impurities commercially available from reference standards providers.4  The method development approach described above will be used to screen columns and solvents to determine suitable method conditions providing separation of the analytes and optimal peak shape.  The integrity of the spectra at low detection levels were evaluated for UV and MS, facilitating peak tracking, purity and identification. In addition, the methodology was compared to reversed-phase results to assess the orthogonality of the two chromatographic techniques.

REFERENCES:
1.        http://www.ftc.gov/opa/2002/12/baxter_wyeth.shtm
2.        SHIDHAYE, S.. Development and Validation of Stability Indicating HPLC Method for Estimation of Metoclopramide Hydrochloride From A Novel Formulation. Journal of Pharmacy Research, UAE, 2, aug. 2009
3.        Lee, H. W., H. Y. Ji, et al. (2009). "Determination of metoclopramide in human plasma using hydrophilic interaction chromatography with tandem mass spectrometry." Journal of Chromatography B 877(18-19): 1716-1720.
4.        LGC Standards


Converging Chromatographic Approaches: 
A Critical Evaluation of Sub-2µm particles and Supercritical CO2 mobile phases
Michael D. Jones
Waters Corporation


Abstract
The use and benefits of sub-2µm chromatographic particles in reversed phase chromatography (RPLC) is well established.   Often, packed column supercritical fluid chromatography (pSFC) is performed using 5µm particles typically with columns of 150-250 mm in length.  Although many users have had success using this approach, limitations in analytical SFC instrument design and column availability have prevented the adoption of small particle pSFC.   However, the use of SFC has seen a resurgence of use within the pharmaceutical industry due to the orthogonality to HPLC as well as its "Green" chemistry aspects [1].   pSFC has been readily adopted for chiral and more recently, achiral separations for purification of compounds.  The driving factors have been decreases in run time; increase in chromatographic efficiencies, less solvent usage and shorter evaporation times and [2-5].   With respect to analytical pSFC, many systems identified in the literature utilize configurations limited by large system volume and extra column volumes affecting the ability to achieve optimal chromatographic performance.  The scope of this work explores the impact of a low dwell volume and low extra column volume instrument design on chromatographic efficiency, resolution, sensitivity for sub 2um particles stationary phases. The advantages of overcoming these instrumental performance limitations will allow a true assessment of pSFC for compatible sub-2µm stationary phases.

 In this presentation, we will evaluate the evolution of supercritical fluid chromatography to the convergence of liquid and gas chromatographic techniques.  We will highlight recent academic research helping to shape our theoretical foundation of knowledge as it pertains to supercritical fluids as mobile phases.  We study column efficiencies using columns packed with different particle size using the latest commercially available instrumentation, UltraPerformance Convergence Chromatography (UPC2).  We will address technical aspects of this new technique such as flow/pressure regimes and the impact of  pressure/temperature drops across the various column configurations.  We will also discuss the implementation of UPC2 for the analysis of small molecule pharmaceutical analysis, lipidomics, and traditional medicine profiling.

REFERENCES:
1.        Harris CM. The SFC comeback. Pharmaceuticals give supercritical fluid chromatography a fighting chance. Anal Chem. 2002 Feb 1;74(3):87A-91A
2.        B. Bolanos, M. Greig, M. Ventura, W. Farrell, C.M. Aurigemma,H. Li, T.L. Quenzer, K. Tivel, J.M.R. Bylund, P. Tran, C. Pham, D.Phillipson, Int. J. Mass Spectrom. 238 (2004) 85.
3.        M. Ventura, W. Farrell, C. Aurigemma, K. Tivel, M. Greig, J. Wheatley,A. Yanovsky, K.E. Milgram, D. Dalesandro, R. DeGuzman, P.Tran, L. Nguyen, L. Chung, O. Gron, C.A. Koch, J. Chromatogr. A
1036 (2004) 7.
4.         T. Wang, M. Barber, I. Hardt, B. Kassel, Rapid Commun. Mass Spectrom. 15 (2001) 2067.
5.         C. White, J. Burnett, J. Chromatogr A. 1074 (2005) 175-185


Biography
Michael Jones is a graduate of the University of Rhode Island obtaining a Bachelors of Science degree in Chemistry in 1998.  His undergraduate research focused on CE analysis of fullerenes and HPLC analysis of nucleotides under the guidance of Phyllis Brown.

Mr. Jones was employed by AstraZeneca in 1998 with responsibilities of QC finished products analysis of various injectables and inhalation suspensions.  His responsibilities increased over the four years with AstraZeneca to include long term stability testing, method transfer, and drug product quality investigations.

Currently, Michael is a senior scientist within the Pharmaceutical Life Sciences Organization at Waters Corporation.  Mr. Jones started in Waters in 2001 and primarily worked to develop automated method development solutions.  Since 2003, Michael's responsibilities shifted to ACQUITY UPLC applications focusing on pharmaceutical impurity analysis covering a wide range of application space related to LC and MS technologies resulting in numerous peer reviewed publications.  His experience with UPLC have facilitated collaborations involving Pharma and their goals to globally adopt UPLC through system effectiveness education. 

Recently, Michael was accepted to King's College for graduate research study focusing on analytical scale sub-2µm packed column supercritical fluid techniques.  The goal of the body of work will be to gain a better understanding of sub-2µm pSFC chromatographic behavior when performing experiments using systems that transcend the limitations of today's SFC technology.  Although small molecule pharmaceutical analysis is intended to define much of the research direction, other areas such as lipid analysis, drugs of abuse, natural products, and traditional medicines will be explored to determine feasibility of sub-2um analytical pSFC as a viable analytical choice within those respective disciplines.