Coupling of Liquid Chromatography with ICP-MS is straightforward. Nebulizers are available that can accept the effluent from standard, µ-HPLC and even nano-HPLC. With all the different possiblities of ion exchange, reverse-phase, affinity, size-exclusion, ion-pairing and any other of the numerous techniques, liquid chromatography offers a very rich tool-box for separation that can be used to separate many of the species of interest.
Liquid chromatography (LC) is an extremely versatile technique with numerous advantages:
- both the stationary phase and the mobile phase may be altered to achieve the desired separation,
- separations can be further enhanced by the addition of additives (chiral additives, ion pair reagents, surfactants) to the mobile phase,
- separations can be enhanced by changing the mobile phase during the analysis (gradient elution),
- an enourmous variety of stationary phases are commercially available based on a rich variety of separation principles such as
- normal phase chromatography (NPC)
- reversed phase chromatography (RPC)
- reversed phase ion pair chromatography (IPC)
- micellar chromatography
- ion exchange chromatography (IEC)
- size exclusion chromatography (SEC).
- Usually, minimum sample preparation is required.
Fig. 1 Coupling of HPLC with ICP-MS
Liquid sample introduction is the standard in ICP-MS, therefore coupling of liquid chromatography with ICP-MS in the simplest form is just the connection of the column outlet with the nebulizer of the sample introduction system via a transfer tubing. It is not a surprise that the hyphenated system resulting from the coupling of liquid chromatography and ICP-MS is the most often used system for speciation analysis related to the ICP-MS detection. About 1/3 of all publications on speciation analysis related to ICP-MS describe the use of LC-ICP-MS.
Fig. 2: Development of publications related to LC-ICP-MS for speciation analysis
LC-ICP-MS has important advantages for speciation analysis:
- Complex chromatograms are reduced to simple "elementograms"
- Quantification of even unknown element species is possible with respect to the detected element due to compound independent sensitivity without the necessity of having standards.
However the hard ionization and atomization power of the plasma source also has some drawbacks:
- All molecular information is lost
- Species characterization is limited to chromatographic retention times
Looking more closely onto the hyphenated system, some details have to be discussed:
- contamination with metallic compounds arising from the chromatographic system (pump, valve, tubing) or the stationary and mobile phase,
- dispersion effects, that means the influence on the sample introduction system on the separation power of the chromatographic system,
- plasma solvent load effects, that means the influence on the mobile phase on the stability and ionization power of the inductively coupled plasma.
Plasma solvent load effects limit the free choice of the mobile phase, since now the mobile phase has to be selected both with respect to the separation task but also with respect to the degree of tolerance by the ICP. Tolerance by the ICP can be enhanced by reducing the flow-rate, which can be achieved by reducing the size-factor of the HPLC system from standard to micro-HPLC or even nano-HPLC. Tolerance of the ICP with nano-liter flow rates is so high that nearly any solvent can be introduced with total consumption nebulizers.
On the other hand, the negative influence of dead-volumes and other poor design factors on the chromatographic separation power is the more pronounced the smaller the flow rate of the system is. Therefore, especially careful design considerations are necessary for nano-HPLC coupling while standard system are less critical.
With respect to the type of MS detection system, one has to consider the transient character of the chromatogtaphic signals. Depending on the number of simultaneous channels that have to be monitored (only one in the simple case of a single element speciation), systems that can either simultaneously collect data on different channels or can do fast sequential measurements will have advantages.
Tutorial material related to HPLC
Animation of Dual Head HPLC Pump
Animation of sample loop HPLC injections
HPLC Trouble-Shooting
IonSource.com: Introduction to Capillary Chromatography
IUPAC: Chromatography Nomenclature and Definitions
LC-GC Europe: Glossary of Liquid-Phase Separation Terms
Prof. Yuri Kazakevich, Prof. H. M. McNair, On-line Textbook on High Performance Liquid Chromatography (HPLC)
UCDAVIS CHEMWIKI: HPLC
Upchurch Scientific: HPLC Introduction
Waters: HPLC - High Performance Liquid Chromatography
Reviews of LC-ICP-MS (newest first)
Maximilian Popp, Stephan Hann, Gunda Koellensperger, Environmental application of elemental speciation analysis based on liquid or gas chromatography hyphenated to inductively coupled plasma mass spectrometry - A review, Anal. Chim. Acta, 668/2 (2010) 114-129. doi: 10.1016/j.aca.2010.04.036
John E. Carr, Adam E Dill, Kaho Kwok, Jon W. Carnahan, G.K. Webster, LC-ICP-MS for Nonmetal Selective Detection of Pharmaceuticals, Curr. Pharma. Anal., 4/4 (2008) 206-214. doi 10.2174/157341208786306234
Kevin A. Francesconi, Michael Sperling, Speciation analysis with HPLC-mass spectrometry: time to take stock, Analyst (London), 130/7 (2005) 998-1001.
DOI: 10.1039/b504485p
Maria Montes-Bayón, K. DeNicola, Joseph A. Caruso, Liquid chromatography-inductively coupled plasma mass spectrometry, J. Chromatogr. A, 1000 (2003) 457-476. doi:10.1016/S0021-9673(03)00527-2
Joanna Szpunar, Ryszard Lobinski, Liquid Chromatography with ICP MS Detection, in: Hyphenated Techniques in Speciation Analysis, RSC, Cambridge, 2003, pp. 53-64.
Bernhard Michalke, The coupling of LC to ICP-MS in element speciation - Part II: Recent trends in application, Trends Anal. Chem. (Pers. Ed.), 21/3 (2002) 154-165. doi:10.1016/S0165-9936(02)00303-5
Bernhard Michalke, The coupling of LC to ICP-MS in element speciation: I. General aspects, Trends Anal. Chem. (Pers. Ed.), 21/2 (2002) 142-153. doi:10.1016/S0165-9936(01)00146-7
Studies on the instrumentation and instrumental parameters of LC-ICP-MS
Lothar Rottmann, Klaus G. Heumann, Development of an on-line isotope dilution technique with HPLC/ICP-MS for the accurate determination of elemental species, Fresenius J. Anal. Chem., 350/4-5 (1994) 221-227. doi: 10.1007/BF00322473
Cristina Rivas, Les Ebdon, Steve J. Hill, Effect of different spray chambers on the determination of organotin compounds by high-performance liquid chromatography - inductively coupled plasma mass spectrometry, J. Anal. At. Spectrom., 11/12 (1996) 1147-1150. DOI: 10.1039/JA9961101147
Warren R.L. Cairns, Les Ebdon, Steve J. Hill, A high performance liquid chromatography - inductively coupled plasma-mass spectrometry interface employing desolvation for speciation studies of platinum in chemotherapy drugs, Fresenius J. Anal. Chem., 355/3-4 (1996) 202-208. doi: 10.1007/s0021663550202
Anders Tangen, Roger Trones, Tyge Greibrokk, Walter Lund, Microconcentric Nebulizer for the Coupling Of Micro Liquid Chromatography and Capillary Zone Electrophoresis With Inductively Coupled Plasma Mass Spectrometry, J. Anal. At. Spectrom., 12/6 (1997) 667-670. doi: 10.1039/a607623h
Mary Kate Donais, How to Interface a Liquid Chromatograph to an Inductively Coupled Plasma Mass Spectrometer for Elemental Speciation Studies, Spectroscopy (Eugene, Oreg.), 13/9 (1998) 30-35.
Clayton B'Hymer, Karen L. Sutton, Joseph A. Caruso, Comparison of four nebulizer-spray chamber interfaces for the high-performance liquid chromatographic separation of arsenic compounds using inductively coupled plasma mass spectrometric detection, J. Anal. At. Spectrom., 13/9 (1998) 855-858. doi: 10.1039/a801645c
Erik H. Larsen, Method optimization and quality assurance in speciation analysis using high-performance liquid chromatography with detection by inductively coupled plasma mass spectrometry, Spectrochim. Acta, Part B, 53/2 (1998) 253-265.
doi: 10.1016/S0584-8547(97)00137-7
B. Do, S. Robinet, D. Pradeau, F. Guyon, Application of central composite designs for optimisation of the chromatographic separation of monomethylarsonate and dimethylarsinate and of selenomethionine and selenite by ion-pair chromatography coupled with plasma mass spectrometric detection, Analyst (London), 126/5 (2001) 594. DOI: 10.1039/b008169h
Enrique G. Yanes, Nancy J. Miller-Ihli, Use of a parallel path nebulizer for capillary-based microseparation techniques coupled with an inductively coupled plasma mass spectrometer for speciation measurements, Spectrochim. Acta, Part B, 59/6 (2004) 883-890. doi:10.1016/j.sab.2004.03.005
Lars Bendahl, Bente Gammelgaard, Sample introduction systems for reversed phase LC-ICP-MS of selenium using large amounts of methanol - Comparison of systems based on membrane desolvation, a spray chamber and direct injection, J. Anal. At. Spectrom., 20/5 (2005) 410-416. DOI: 10.1039/b415717f
Lars Bendahl, Stefan Stürup, Bente Gammelgaard, Steen Honoré Hansen, Ultra-Performance LC-ICP-MS - a fast technique for speciation analysis, J. Anal. At. Spectrom., 20/11 (2005) 1287-1289. DOI: 10.1039/b508653a
Zs. Stefánka, Gunda Koellensperger, Gerhard Stingeder, Stephan Hann, Down-scaling narrowbore LC-ICP-MS to capillary LC-ICP-MS: a comparative study of different introduction systems, J. Anal. At. Spectrom., 21/1 (2006) 86-89. DOI: 10.1039/b511629e
Yasumitsu Ogra, Development of Miniaturized HPLC-ICP-MS for Speciation of Bio-Trace Elements, Biomed. Res. Trace Elem., 19/1 (2008) 34-42. available at: http://www.jstage.jst.go.jp/article/brte/19/1/34/_pdf
Kirk E. Lokits, Patrick A. Limbach, Joseph A. Caruso, Interfaces for capillary LC with ICPMS detection: A comparison of nebulizers/spray chamber configurations, J. Anal. At. Spectrom., 24/4 (2009) 528-534. DOI: 10.1039/b820121h
Kenneth Neubauer, Advantages and Disadvantages of Different Column Types for Speciation Analysis by LC-ICP-MS, Spectroscopy, 24/11 (2009) 30-33. online available here
Daniel Pröfrock, Hyphenation of capillary-LC with ICP-MS and parallel on-line micro fraction collection for MALDI-TOF-MS analysis - complementry tools for protein phosphorylation analysis, J. Anal. At. Spectrom., 25/3 (2010) 33-344. doi: 10.1039/b921145d
Standard operating procedures and methods for LC-ICP-MS
EPA Method 321.8 - Determination of Bromate in Drinking Waters by Ion Chromatography Inductively Coupled Plasma - Mass Spectrometry
Instrument manufacturer's application notes
Agilent Technologies: Handbook of Hyphenated ICP-MS Applications
Agilent Technologies: #5989-7073EN: Determination of Organic and Inorganic Selenium Species Using HPLC-ICP-MS
Agilent Technologies: #5989-3572EN: Determination of Methyl Mercury in Water and Soil by HPLC-ICP-MS
Agilent Technologies: #5989-2481EN: Ion Chromatography (IC) ICP-MS for Chromium Speciation in Natural Samples
Agilent Technologies: #5988-9893EN: Fast and Accurate Determination of Arsenobetaine (AsB) in Fish Tissues using HPLC-ICP-MS
Agilent Technologies: #5988-6697EN: Comparison of GC-ICP-MS and HPLC-ICP-MS for the Analysis of Organotin Compounds
Agilent Technologies: #5988-4332EN: Technical
Features of ICP-MS Plasma Chromatographic Software
Agilent Technologies: #5968-3050EN: Speciation of Arsenic Compounds in Urine of Dimethylarsinic Acid Orally Exposed Rat by Using IC-ICP-MS
PerkinElmer: #D-6736: Speciation of Five Arsenic Compounds in Urine by HPLC ICP-MS
PerkinElmer: #D-6780: Chromium Speciation in Water by HPLC ICP-MS
PerkinElmer: #D-7303: Advances in Bromine Speciation by HPLC/ICP-MS
Thermo Scientific: Poster WPC 01/2008: Shona McSheehy, Tomoko Oki, Julian Wills, Meike Hamester, Comparison of GC-ICP-MS and HPLC-ICP-MS for Speciation of Mercury in Blood
Thermo Scientific: #30012: High Sensitivity Arsenic Speciation: HPLC Sector Field ICP-MS
Thermo Scientific: #30076: Simultaneous Phosphorus and Sulfur Speciation by HPLC Interfaced with High Resolution ICP-MS
Thermo Scientific: #40653: Determination of Organo-Arsenic Species in Seaweed Extracts using HPLC-ICP-MS
Thermo Scientific: #40720: Determination of Arsenic Species in Urine Using HPLC Coupled with X Series ICP-MS
Thermo Scientific: #40741: Speciation of Arsenic in Fish Tissues using HPLC coupled with X SeriesII ICP-MS
Thermo Scientific: #40745: Determination of Selenomethionine in Nutritional Supplements using HPLC coupled to the XSeriesII ICP-MS with CCT Thermo Scientific: #40782: The Speciation of Mercury in Biomedical and Environmental Samples using HPLC coupled to the XSeries 2 ICP-MS Thermo Scientific: #40807: Chromium Speciation in Cement Extracts and Airborne Particulates using PLC Coupled with the XSERIES 2 ICP-MS
Thermo Scientific: #40837: The Determination of Trivalent and Hexavalent Chromium in Mineral and Spring Water using HPLC Coupled to the XSeriesII ICP-MS with CCT
Varian: #29: Low level speciation of chromium in drinking waters using LC-ICP-MS (pdf, 146 KB)
Varian: #27: The speciation of Se(IV) and Se(VI) using the Varian 820-MS (pdf, 146 KB)
Varian: #26: Speciation of Arsenic by LC-ICP-MS (pdf, 461 KB)
Varian: # 9: HPLC-ICP-MS, Metal Speciation in Tea using Gradient Elution
HPLC Maintenance and Trouble-Shooting
HPLC Trouble-Shooting
Agilent 1100 Series LC and LC/MS Maintenance Videos (only for registered users) EVISA Database system Journals Database: Journals related to Liquid Chromatography
Company Database: Professional Organizations relelated to Chromatography
Company Database: Manufacturers providing LC-ICP-MS systems
Instrument Database: LC-ICP-MS coupling kits
Instrument Database: HPLC Autosampler
Instrument Database: HPLC pumps
EVISA link pages
Resources related to analytical sciences
Resources related to mass spectrometry
Resources related to Chromatography
Resources related to quality assurance/quality control
Other web resources:
Agilent Technologies: Elemental Speciation Measurement - Combining Chromatography and ICP-MS
PerkinElmer Instruments: Online Seminar: Applied Speciation Analysis in Environmental and Clinical Studies
Thermo Scientific: Complete Speciation Solutions
Royal Society of Chemistry > Analytical Methods Commitee > Report of the Instrumental Criteria Subcommittee> Criteria for the Selection of Instruments: Part IX (1997): Instrumentation for High-performance Liquid Chromatography
News related to LC-ICP-MS for speciation analysis (newest first)
EVISA News, October 7, 2007: Agilent Technologies publishes Handbook of Hyphenated ICP-MS Applications
EVISA News, January 24, 2007: Agilent Technologies joins University of Cincinnati to study impact of metal species in biology and the environment
EVISA News, October 18, 2006: Speciation analysis by LC-ICP-MS finds new application area in clinical chemistry: Ceruloplasmin
EVISA News, September 7, 2006: New Agilent HPLC column for routine determination of arsenic species in human urine by HPLC-ICP-MS
EVISA News, January 25, 2006: A new concentric nebulizer for ICP sample introduction at nL-sample flow rates
EVISA News, October 20, 2004: Thermo Electron and Pau University Establish New Center of Excellence for Elemental Speciation Analysis
EVISA News, August 26, 2004: Ion chromatography and ICP-MS determination of arsenic species in marine samples
Frost & Sullivan, January 6, 2004: Hyphenated Techniques Breathing New Life into the Analytical Instrumentation Industry
Further chapters:
Chapter 1: Tools for elemental speciation
Chapter 2: ICP-MS - A versatile detection system for speciation analysis
Chapter 3: LC-ICP-MS - The most often used hyphenated system for speciation analysis
Chapter 4: GC-ICP-MS
Chapter 5: CE-ICP-MS for speciation analysis
Chapter 6: ESI-MS: The tool for the identification of species
last time modified: November 8, 2011
