Abstract
One-dimensional liquid chromatography (1D-LC) is not always capable of efficiently separating complex samples. This drawback is not solely due to the lack of column efficiency, but is mainly due to insufficient selectivity and the need to separate the analytes of interest with orthogonal retention mechanisms. In this regard, two-dimensional liquid chromatography (2D-LC) is currently attracting much interest for its markedly higher resolving power compared to one-dimensional separation. In this work, three applications of 2D-LC from the pharmaceutical industry are presented with the goal not only to increase peak capacity, but also to support investigations. In the first application, the retention times of peaks of interest are matched under different mobile phase conditions for the purpose of transferring the method from a mass spectrometry (MS) incompatible buffer to an MS compatible buffer. The second application includes developing a method for simultaneous detection and quantitation of degradants and aggregates in a biologics and small molecule combination product. The third application supports method development by confirming the purity of separated peaks using orthogonal separation conditions in the first and second dimensions and to investigate mass balance issues where some peaks are expected to elute in the solvent front.
Similar content being viewed by others
References
Haidar Ahmad IA, Soliven A, Allen RC, Filgueira M, Carr PW (2015) J Chromatogr A 1386:31–38
Allen RC, Barnes BB, Haidar Ahmad IA, Filgueira MR, Carr PW (2014) J Chromatogr A 1361:169–177
Stoll DR, Cohen JD, Carr PW (2006) J Chromatogr A 1122:123–137
Li X, Stoll DR, Carr PW (2008) Anal Chem 81:845–850
Potts LW, Stoll DR, Li X, Carr PW (2010) J Chromatogr A 1217:5700–5709
Pirok BWJ, Gargano AFG, Schoenmakers PJ (2017) J Sep Sci. https://doi.org/10.1002/jssc.201700863
Scoparo CT, de Souza LM, Dartora N, Sassaki GL, Gorin PAJ, Iacomini M (2012) J Chromatogr A 1222:29–37
Li Y, Gu C, Gruenhagen J, Zhang K, Yehl P, Chetwyn NP, Medley CD (2015) J Chromatogr A 1393:81–88
Dugo P, Fawzy N, Cichello F, Cacciola F, Donato P, Mondello L (2013) J Chromatogr A 1278:46–53
Wang S, Li J, Shi X, Qiao L, Lu X, Xu G (2013) J Chromatogr A 1321:65–72
Montero L, Herrero M, Ibáñez E, Cifuentes A (2013) J Chromatogr A 1313:275–283
Kittlaus S, Schimanke J, Kempe G, Speer K (2013) J Chromatogr A 1283:98–109
Vanhoenacker G, Vandenheede I, David F, Sandra P, Sandra K (2015) Anal Bioanal Chem 407:355–366
Matejicek D (2012) J Chromatogr A 1231:52–58
Petersson P, Haselmann K, Buckenmaier S (2016) J Chromatogr A 1468:95–101
Zhang K, Li Y, Tsang M, Chetwyn NP (2013) J Sep Sci 36:2986–2992
Maloney DRSTD (2017) Recent advances in two-dimensional liquid chromatography for pharmaceutical and biopharmaceutical analysis. LCGC North America 35(9):680–687
Striegel A, Yau WW, Kirkland JJ, Bly DD (2009) Modern size-exclusion liquid chromatography: practice of gel permeation and gel filtration chromatography. Wiley, Hoboken
Boone MA, Striegel AM (2006) Macromolecules 39:4128–4131
Caltabiano AM, Foley JP, Striegel AM (2018) J Chromatogr A 1532:161–174
Lee C, Zang J, Cuff J, McGachy N, Natishan TK, Welch CJ, Helmy R, Bernardoni F (2013) Chromatographia 76:5–11
Hajslova J, Zrostlikova J (2003) J Chromatogr A 1000:181–197
Lincoln D, Fell AF, Anderson NH, England D (1992) J Pharm Biomed Anal 10:837–844
Keller HR, Massart DL (1991) Anal Chim Acta 246:379–390
Wiberg K, Andersson M, Hagman A, Jacobsson SP (2004) J Chromatogr A 1029:13–20
Ahmad IAH, Hrovat F, Soliven A, Clarke A, Boswell P, Tarara T, Blasko A (2017) Chromatographia 80:1143–1159
Haidar Ahmad IA (2017) Chromatographia 80:705–730
Dolan J (2011) Selectivity in reversed-phase LC separations, part III: column-type selectivity. In: LCGC North America, pp 236–244
Snyder LR, Dolan JW, Carr PW (2004) J Chromatogr A 1060:77–116
Venkatramani CJ, Girotti J, Wigman L, Chetwyn N (2014) J Sep Sci 37:3214–3225
Zhang K, Wang J, Tsang M, Wigman L, Chetwyn NP (2013) Am Pharm Rev 16:39–44
Wilson NS, Nelson MD, Dolan JW, Snyder LR, Wolcott RG, Carr PW (2002) J Chromatogr A 961:171–193
Acknowledgements
The authors would like to thank the Agilent for the provision of loan instrument. We would like also to acknowledge Bob Giuffre, Kritsin Swanson, and Lisa Zhang from Agilent Technologies for their helpful suggestions and discussions. We would like to thank members of the Analytical Network at Novartis Pharmaceuticals for their helpful discussions.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors have no potential conflicts of interest to declare.
Human and animal rights statement
The research presented here did not involve human participants and/or animals.
Rights and permissions
About this article
Cite this article
Ahmad, I.A.H., Blasko, A., Clarke, A. et al. Two-Dimensional Liquid Chromatography (2D-LC) in Pharmaceutical Analysis: Applications Beyond Increasing Peak Capacity. Chromatographia 81, 401–418 (2018). https://doi.org/10.1007/s10337-018-3474-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10337-018-3474-8