ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Loop-Based Multiple Heart-Cutting Two-Dimensional Liquid Chromatography for Target Analysis in Complex Matrices

View Author Information
Dow Deutschland Anlagengesellschaft mbH, Analytical Technology Center (ATC), Industriestr. 1, 77836 Rheinmuenster, Germany
Agilent Technologies, Research and Development, Hewlett-Packard-Str. 8, 76337 Waldbronn, Germany
*E-mail: [email protected]. Phone: +49 7227 913699.
Cite this: Anal. Chem. 2015, 87, 10, 5310–5317
Publication Date (Web):April 21, 2015
Copyright © 2015 American Chemical Society

    Article Views





    Other access options


    Abstract Image

    Loop-based multiple heart-cutting (MHC) two-dimensional liquid chromatography (2D-LC) is presented as a solution to quantify target components in complex matrices, such as additives in polymers, at very high chromatographic resolution. The determination of hexabromocyclododecane (HBCD) in polystyrene (PS) is described. One dimensional (1D) LC analysis with UV detection did not allow quantitation of the main isomers of HBCD due to peak overlap with polymer components. MHC 2D-LC analysis provided the separation power, accuracy, and repeatability needed for quantitative analysis of the additives of interest. Heart-cuts from peaks of the 1D-chromatogram or entire regions of interest are sampled into loops, where they remain parked until their sequential reinjection onto the second dimension (2D) column. A column set consisting of phenyl (1D) and C18 (2D) stationary phases gave baseline separation in 2D between HBCD and PS background. Linearity for spiked polymer samples was achieved over a range of 0.02–1.00 wt % HBCD relative to the amount of polymer. The limit of quantitation was estimated at 0.01 wt % HBCD in PS. A peak area RSD of 0.7% obtained for ten replicates of a real sample demonstrated excellent repeatability of the analysis. MHC 2D-LC is an elegant solution for quantitative analyses of difficult-to-separate samples when conventional 1D separation fails.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Cited By

    This article is cited by 58 publications.

    1. Ryan Karongo, Jeannie Horak, Michael Lämmerhofer. Comprehensive Online Reversed-Phase × Chiral Two-Dimensional Liquid Chromatography-Mass Spectrometry with Data-Independent Sequential Window Acquisition of All Theoretical Fragment-Ion Spectra-Acquisition for Untargeted Enantioselective Amino Acid Analysis. Analytical Chemistry 2022, 94 (49) , 17063-17072.
    2. Mireia Rodriguez-Zubiri, François-Xavier Felpin. Analytical Tools Integrated in Continuous-Flow Reactors: Which One for What?. Organic Process Research & Development 2022, 26 (6) , 1766-1793.
    3. Jelle De Vos, Magali Dams, Ken Broeckhoven, Gert Desmet, Burkhard Horstkotte, Sebastiaan Eeltink. Prototyping of a Microfluidic Modulator Chip and Its Application in Heart-Cut Strong-Cation-Exchange-Reversed-Phase Liquid Chromatography Coupled to Nanoelectrospray Mass Spectrometry for Targeted Proteomics. Analytical Chemistry 2020, 92 (3) , 2388-2392.
    4. Aogu Furusho, Reiko Koga, Takeyuki Akita, Masashi Mita, Tomonori Kimura, Kenji Hamase. Three-Dimensional High-Performance Liquid Chromatographic Determination of Asn, Ser, Ala, and Pro Enantiomers in the Plasma of Patients with Chronic Kidney Disease. Analytical Chemistry 2019, 91 (18) , 11569-11575.
    5. Bob W. J. Pirok, Dwight R. Stoll, Peter J. Schoenmakers. Recent Developments in Two-Dimensional Liquid Chromatography: Fundamental Improvements for Practical Applications. Analytical Chemistry 2019, 91 (1) , 240-263.
    6. Dawei Qi, Ting Fei, Hong Liu, Heming Yao, Da Wu, and Baizhan Liu . Development of Multiple Heart-Cutting Two-Dimensional Liquid Chromatography Coupled to Quadrupole-Orbitrap High Resolution Mass Spectrometry for Simultaneous Determination of Aflatoxin B1, B2, G1, G2, and Ochratoxin A in Snus, a Smokeless Tobacco Product. Journal of Agricultural and Food Chemistry 2017, 65 (45) , 9923-9929.
    7. Dwight R. Stoll, Konstantin Shoykhet, Patrik Petersson, and Stephan Buckenmaier . Active Solvent Modulation: A Valve-Based Approach To Improve Separation Compatibility in Two-Dimensional Liquid Chromatography. Analytical Chemistry 2017, 89 (17) , 9260-9267.
    8. Chandan L. Barhate, Erik L. Regalado, Nathan D. Contrella, Joon Lee, Junyong Jo, Alexey A. Makarov, Daniel W. Armstrong, and Christopher J. Welch . Ultrafast Chiral Chromatography as the Second Dimension in Two-Dimensional Liquid Chromatography Experiments. Analytical Chemistry 2017, 89 (6) , 3545-3553.
    9. Shuangyuan Wang, Xianzhe Shi, and Guowang Xu . Online Three Dimensional Liquid Chromatography/Mass Spectrometry Method for the Separation of Complex Samples. Analytical Chemistry 2017, 89 (3) , 1433-1438.
    10. Dwight R. Stoll and Peter W. Carr . Two-Dimensional Liquid Chromatography: A State of the Art Tutorial. Analytical Chemistry 2017, 89 (1) , 519-531.
    11. Yilan Ouyang, Yangyang Zeng, Yinxiu Rong, Yue Song, Lv Shi, Bo Chen, Xinlei Yang, Naiyu Xu, Robert J. Linhardt, and Zhenqing Zhang . Profiling Analysis of Low Molecular Weight Heparins by Multiple Heart-Cutting Two Dimensional Chromatography with Quadruple Time-of-Flight Mass Spectrometry. Analytical Chemistry 2015, 87 (17) , 8957-8963.
    12. Xiao-yan Xu, Mei-ting Jiang, Yu Wang, He Sun, Qi Jing, Xiao-hang Li, Bei Xu, Ya-dan Zou, He-shui Yu, Zheng Li, De-an Guo, Wen-zhi Yang. Multiple heart-cutting two-dimensional liquid chromatography/charged aerosol detector assay of ginsenosides for quality evaluation of ginseng from diverse Chinese patent medicines. Journal of Chromatography A 2023, 1708 , 464344.
    13. Ryan Karongo, Jeannie Horak, Michael Lämmerhofer. Comprehensive reversed‐phase×chiral two‐dimensional liquid chromatography coupled to quadrupole‐time‐of‐flight tandem mass spectrometry with post‐first dimension flow splitting for untargeted enantioselective amino acid analysis. Journal of Separation Science 2023, 46 (18)
    14. Ziqing Lin, Qinggang Wang, Yiyang Zhou, Jonathan G. Shackman. Trapping mode two-dimensional liquid chromatography for quantitative low-level impurity enrichment in pharmaceutical development. Journal of Chromatography A 2023, 1700 , 464043.
    15. Irene Caño‐Carrillo, Bienvenida Gilbert‐López, Lidia Montero, Ana B. Martínez‐Piernas, Juan F. García‐Reyes, Antonio Molina‐Díaz. Comprehensive and heart‐cutting multidimensional liquid chromatography–mass spectrometry and its applications in food analysis. Mass Spectrometry Reviews 2023, 1614
    16. Irene Caño‐Carrillo, Bienvenida Gilbert‐López, Lidia Montero, Ana B. Martínez‐Piernas, Juan F. García‐Reyes, Antonio Molina‐Díaz. Comprehensive and heart‐cutting multidimensional liquid chromatography‐mass spectrometry and its applications in food analysis. Mass Spectrometry Reviews 2023, 1614
    17. Ina Varfaj, Simone Moretti, Federica Ianni, Carolina Barola, Ghaid W. A. Abualzulof, Andrea Carotti, Lina Cossignani, Roberta Galarini, Roccaldo Sardella. Multiple Heart-Cutting Two-Dimensional HPLC-UV Achiral–Chiral Analysis of Branched-Chain Amino Acids in Food Supplements under Environmentally Friendly Conditions. Separations 2023, 10 (1) , 45.
    18. Ruhul Amin, Faruk Alam, Biplab Kumar Dey, Jithendar Reddy Mandhadi, Talha Bin Emran, Mayeen Uddin Khandaker, Sher Zaman Safi. Multidimensional Chromatography and Its Applications in Food Products, Biological Samples and Toxin Products: A Comprehensive Review. Separations 2022, 9 (11) , 326.
    19. Jiawen CHENG, Jiping MA, Shuang LI, Yong TIAN. Progress in sample pretreatment and detection methods of hexabromocyclododecanes. Chinese Journal of Chromatography 2022, 40 (10) , 872-881.
    20. Yunle Huang, Rui Min Vivian Goh, Aileen Pua, Shao Quan Liu, Kim Huey Ee, Benjamin Lassabliere, Bin Yu. Characterisation of catechins and their oxidised derivatives in Ceylon tea using multi-dimensional liquid chromatography and high-resolution mass spectrometry. Journal of Chromatography A 2022, 1682 , 463477.
    21. Matthias Olfert, Stefanie Bäurer, Marc Wolter, Stephan Buckenmaier, Edmundo Brito-de la Fuente, Michael Lämmerhofer. Comprehensive profiling of conjugated fatty acid isomers and their lipid oxidation products by two-dimensional chiral RP×RP liquid chromatography hyphenated to UV- and SWATH-MS-detection. Analytica Chimica Acta 2022, 1202 , 339667.
    22. Yunle Huang, Vivian Rui Min Goh, Aileen Pua, Shao Quan Liu, Kim Huey Ee, Benjamin Lassabliere, Bin Yu. Characterisation of Catechins and Their Oxidised Derivatives in Ceylon Tea Using Multi-Dimensional Liquid Chromatography and High-Resolution Mass Spectrometry. SSRN Electronic Journal 2022, 1362
    23. Mingzhe Sun, Peter Schoenmakers. A compound post-column re-focusing approach in supercritical fluid chromatography. Journal of Chromatography A 2021, 1660 , 462642.
    24. Amanda Suárez Fernández, Pablo Rodríguez-González, Lydia Álvarez, Montserrat García, Héctor González Iglesias, J. Ignacio García Alonso. Multiple heart-cutting two dimensional liquid chromatography and isotope dilution tandem mass spectrometry for the absolute quantification of proteins in human serum. Analytica Chimica Acta 2021, 1184 , 339022.
    25. Ryan Karongo, Min Ge, Christian Geibel, Jeannie Horak, Michael Lämmerhofer. Enantioselective multiple heart cutting online two-dimensional liquid chromatography-mass spectrometry of all proteinogenic amino acids with second dimension chiral separations in one-minute time scales on a chiral tandem column. Analytica Chimica Acta 2021, 1180 , 338858.
    26. Dwight R. Stoll, Ph.D.. 2 D ‐ HPLC – Method Development for Successful Separations. 2021, 1-21.
    27. Guangying Sun, Munire Abuduaini, Guliqire Adili, Yongxin Zhao, Haji Akber Aisa. Dual-tautomerism separation method based on asymmetric transformation: Gram-scale preparation of high-purity punicalagin from pomegranate peel wastes. Journal of Chromatography A 2021, 1651 , 462281.
    28. Jianmin LI, Yue ZHUO, Yida ZHANG, Na LI, Jianlin WU. Size exclusion-reverse liquid column chromatography-mass spectrometry and its application in the identification of post-translationally modified proteins in rat kidney. Chinese Journal of Chromatography 2021, 39 (1) , 87-95.
    29. Jee Seong Kwak, Nour Bizarri, Sepideh Sharif, Wenyao Peter Zhang, Debasis Mallik, Michael G. Organ. The Synthesis of Warfarin Using a Reconfigurable‐Reactor Platform Integrated to a Multiple‐Variable Optimization Tool. Chemistry – A European Journal 2020, 26 (67) , 15505-15508.
    30. Ryan Karongo, Tohru Ikegami, Dwight R. Stoll, Michael Lämmerhofer. A selective comprehensive reversed-phase×reversed-phase 2D-liquid chromatography approach with multiple complementary detectors as advanced generic method for the quality control of synthetic and therapeutic peptides. Journal of Chromatography A 2020, 1627 , 461430.
    31. Christian P. Haas, Simon Biesenroth, Stephan Buckenmaier, Tom van de Goor, Ulrich Tallarek. Automated generation of photochemical reaction data by transient flow experiments coupled with online HPLC analysis. Reaction Chemistry & Engineering 2020, 5 (5) , 912-920.
    32. Donatella Nardiello, Maria Teresa Melfi, Carla Pignatelli, Diego Centonze. Enhancing online protein isolation as intact species from soy flour samples by actively modulated two-dimensional liquid chromatography (2D-LC). Journal of Pharmaceutical and Biomedical Analysis 2020, 179 , 112976.
    33. Wangjie Lv, Xianzhe Shi, Shuangyuan Wang, Guowang Xu. Multidimensional liquid chromatography-mass spectrometry for metabolomic and lipidomic analyses. TrAC Trends in Analytical Chemistry 2019, 120 , 115302.
    34. Canan Armutcu, Erdoğan Özgür, Tunca Karasu, Engin Bayram, Lokman Uzun, Mehmet Emin Çorman. Rapid Analysis of Polycyclic Aromatic Hydrocarbons in Water Samples Using an Automated On-line Two-Dimensional Liquid Chromatography. Water, Air, & Soil Pollution 2019, 230 (10)
    35. Gert Desmet, Ken Broeckhoven. Extra-column band broadening effects in contemporary liquid chromatography: Causes and solutions. TrAC Trends in Analytical Chemistry 2019, 119 , 115619.
    36. Peilin Yang, Wei Gao, Tianlan Zhang, Matthias Pursch, Jim Luong, Wesley Sattler, Anurima Singh, Scott Backer. Two‐dimensional liquid chromatography with active solvent modulation for studying monomer incorporation in copolymer dispersants. Journal of Separation Science 2019, 42 (17) , 2805-2815.
    37. Yan Zhou, Haobo Zhang, Xianying Wang, Dawei Qi, Wenbo Gu, Da Wu, Baizhan Liu. Development of a heart-cutting supercritical fluid chromatography-high-performance liquid chromatography coupled to tandem mass spectrometry for the determination of four tobacco-specific nitrosamines in mainstream smoke. Analytical and Bioanalytical Chemistry 2019, 411 (13) , 2961-2969.
    38. Zheng Wang, Ting-Ting Xie, Xia Yan, Shi Xue, Jing-Wei Chen, Zhen Wu, Ying-Kun Qiu. Gradual Gradient Two-Dimensional Preparative Liquid Chromatography System for Preparative Separation of Complex Natural Products. Chromatographia 2019, 82 (2) , 543-552.
    39. Yingzhuang Chen, Yajun Wu, Xiaoming Liu, Bowen Li, Deping Hu, Si Huang, Ming Ma, Bo Chen. Pulsed elution modulation for on-line comprehensive two-dimensional liquid chromatography coupling reversed phase liquid chromatography and hydrophilic interaction chromatography. Journal of Chromatography A 2019, 1583 , 98-107.
    40. Gino Groeneveld, Melissa N. Dunkle, Marian Rinken, Andrea F.G. Gargano, Ayako de Niet, Matthias Pursch, Edwin P.C. Mes, Peter J. Schoenmakers. Characterization of complex polyether polyols using comprehensive two-dimensional liquid chromatography hyphenated to high-resolution mass spectrometry. Journal of Chromatography A 2018, 1569 , 128-138.
    41. Yang Yang, Yongmin Zhang, Chong Wei, Jing Li, Wenji Sun. Silver ion chromatography for peak resolution enhancement: Application to the preparative separation of two sesquiterpenes using online heart-cutting LC-LC technique. Talanta 2018, 187 , 252-258.
    42. Jucai Xu, Lin Zheng, Lianzhu Lin, Baoguo Sun, Guowan Su, Mouming Zhao. Stop-flow reversed phase liquid chromatography × size-exclusion chromatography for separation of peptides. Analytica Chimica Acta 2018, 1018 , 119-126.
    43. Matthias Pursch, Antje Wegener, Stephan Buckenmaier. Evaluation of active solvent modulation to enhance two-dimensional liquid chromatography for target analysis in polymeric matrices. Journal of Chromatography A 2018, 1562 , 78-86.
    44. Joe M. Davis, Dwight R. Stoll. Likelihood of total resolution in selective comprehensive two-dimensional liquid chromatography with parallel processing: Simulation and theory. Journal of Chromatography A 2018, 1537 , 43-57.
    45. C.J. Venkatramani, Shu Rong Huang, Mohammad Al-Sayah, Ila Patel, Larry Wigman. High-resolution two-dimensional liquid chromatography analysis of key linker drug intermediate used in antibody drug conjugates. Journal of Chromatography A 2017, 1521 , 63-72.
    46. Jucai Xu, Dongxiao Sun-Waterhouse, Chaoying Qiu, Mouming Zhao, Baoguo Sun, Lianzhu Lin, Guowan Su. Additional band broadening of peptides in the first size-exclusion chromatographic dimension of an automated stop-flow two-dimensional high performance liquid chromatography. Journal of Chromatography A 2017, 1521 , 80-89.
    47. Samuel H. Yang, Jenny Wang, Kelly Zhang. Validation of a two-dimensional liquid chromatography method for quality control testing of pharmaceutical materials. Journal of Chromatography A 2017, 1492 , 89-97.
    48. Abasha Williams, Erik K. Read, Cyrus D. Agarabi, Scott Lute, Kurt A. Brorson. Automated 2D-HPLC method for characterization of protein aggregation with in-line fraction collection device. Journal of Chromatography B 2017, 1046 , 122-130.
    49. Matthias Pursch, Paul Lewer, Stephan Buckenmaier. Resolving Co-Elution Problems of Components in Complex Mixtures by Multiple Heart-Cutting 2D-LC. Chromatographia 2017, 80 (1) , 31-38.
    50. Sebastiaan Eeltink, Jelle De Vos. Ultra-High-Pressure Liquid Chromatography ☆. 2017
    51. Dwight R. Stoll. Introduction to Two-Dimensional Liquid Chromatography—Theory and Practice. 2017, 227-286.
    52. Marta Kula, Daniel Głód, Mirosława Krauze-Baranowska. Application of on-line and off-line heart-cutting LC in determination of secondary metabolites from the flowers of Lonicera caerulea cultivar varieties. Journal of Pharmaceutical and Biomedical Analysis 2016, 131 , 316-326.
    53. Patrik Petersson, Kim Haselmann, Stephan Buckenmaier. Multiple heart-cutting two dimensional liquid chromatography mass spectrometry: Towards real time determination of related impurities of bio-pharmaceuticals in salt based separation methods. Journal of Chromatography A 2016, 1468 , 95-101.
    54. Xia Yan, Li-Juan Wang, Zhen Wu, Yun-Long Wu, Xiu-Xiu Liu, Fang-Rong Chang, Mei-Juan Fang, Ying-Kun Qiu. New on-line separation workflow of microbial metabolites via hyphenation of analytical and preparative comprehensive two-dimensional liquid chromatography. Journal of Chromatography B 2016, 1033-1034 , 1-8.
    55. Maraíssa Silva Franco, Rodrigo Nogueira Padovan, Bruno Henrique Fumes, Fernando Mauro Lanças. An overview of multidimensional liquid phase separations in food analysis. ELECTROPHORESIS 2016, 37 (13) , 1768-1783.
    56. Xue Qiao, Qi Wang, Wei Song, Yi Qian, Yao Xiao, Rong An, De-an Guo, Min Ye. A chemical profiling solution for Chinese medicine formulas using comprehensive and loop-based multiple heart-cutting two-dimensional liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Journal of Chromatography A 2016, 1438 , 198-204.
    57. Chunlei Wang, Sike Chen, John A. Brailsford, Aaron P. Yamniuk, Adrienne A. Tymiak, Yingru Zhang. Characterization and quantification of histidine degradation in therapeutic protein formulations by size exclusion-hydrophilic interaction two dimensional-liquid chromatography with stable-isotope labeling mass spectrometry. Journal of Chromatography A 2015, 1426 , 133-139.
    58. Dwight R Stoll. Recent advances in 2D-LC for bioanalysis. Bioanalysis 2015, 7 (24) , 3125-3142.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Your Mendeley pairing has expired. Please reconnect