Chip technology, nanoliters and picoliters – miniaturization of (bio)analytical chemistry methods Andreas Manz I S A S INSTITUTE FOR ANALYTICAL SCIENCES Dortmund and Berlin
organization Prof.Dr.Andreas Manz Prof.Dr.Kay Niemax head miniaturization proteomics metabolomics spectroscopy materials PD Dr.Joachim Franzke Dr.Norbert Jakubow ski Dr.Jörg I. Baumbach PD Dr.Volker K. Deckert Dr.Roland Hergenröder head a.i. head head head head micro plasmas transcription profiling volatile metabolites GC-IMS nano raman synchrotron XRF (PD Dr.Joachim Franzke) Prof. Dr. Philip Day (Dr.Jörg I. Baumbach) (PD Dr.Volker K. Deckert) Alex von Bohlen microfluidic separations ICP-MS molecular imaging functional biotechnology diode laser AS femtosecond laser ablatio (Prof.Dr.Andreas Manz) (Dr.Norbert Jakubow ski) Prof. Dr. Andreas Schmid (Prof.Dr.Kay Niemax) (Prof.Dr.Kay Niemax) x-ray and neutron sources echelle spectrometers* IR ellipsometry* Prof.Dr.Eduardo Greaves Dr.Helmut Becker-Ross PD Dr.Norbert Esser * Berlin-Adlershof
What do we have now? [example 1] • Electrophoresis chips - Caliper, Agilent, Predicant, Hitachi, Shimadzu etc. • mainly used for DNA fragment sizing • protein separations
Mass spec spraying needle, needle assembly & fittings Nano LC Column Enrichment column, capillaries, fittings, frits HV ESI contact RF tag chip based LC/MS by Agilent courtesy of Tom A.van de Goor, Agilent, Santa Clara CA
What do we have now? [example 2] • Reactor chips - Upchurch, etc. • Mainly used for solvent gradients in chromatography • Chemical synthesis • Bioassays
N+ H H R1 R2 Cl- H H O MeOH N+ Cl- H2 O N R3 R4 R2 R1 C R1 N R2 N R4 R3 H2 O R1 N R2 N R4 R3 O R1/R2 = -CH2 (CH2 )3 CH2 - Piperidine hydrochloride + + Piperidinium cation + R3/R4 = -CH2 (CH2 )4 CH2 - Cyclohexyl isocyanide Nitrilium intermediate -Dialkylacetamide Formaldehyde N-Cyclohexyl-2-piperidin-1-yl-acetamide (1) (2) (3) (4) (5) (6) Multicomponent Chemistries: The Ugi Reaction 0oC
10 mL/min (24sec) 40 mL/min (6sec) His68 Tyr59 N N N N A A A A M.Kakuta, D.A.Jayawickrama, A.M.Wolters, A.Manz, J.V.Sweedler, Anal.Chem. 75, 956-960 (2003)
Br - (MeO)3 Ph P + Ph(OMe)3 (MeO)3 Ph N H NH2 O R H O + Br - (MeO)3 Ph P + Ph(OMe)3 (MeO)3 Ph N H N O H R Acetic acid, 30 mins, sonicate Aldehyde 4-Hydrazino-4-oxobutyl-TMPP+Br- Hydrazino derivative of aldehyde Reaction of Aldehydes (and ketones) with 4-hydrazino-4-oxobutyl-TMPP
Gradient µLC-chip-MS of Amines – isotope labeling analysis Methylheptylamine Methyloctylamine Methyldecylamine Methylhexylamine Products Accurate Mass Difference Isotopes: 2.012 Only characteristic pattern is necessary, not molecular weight
Sequential DNA hybridization • Inject small volume plugs of probe DNA oligomers • Mix within ms with target DNA • Observe hybridization reaction as the plug moves downstreams
Influence of DNA Sequence on Fluorescence Levels Order: matching, 1 mismatch, 2 mismatches, 5 mismatches Order: matching, 1 mismatch, 2 mismatches Sequence-dependent responses from two different experiments.
IEF chip • 36 x 20 um inlet channels • 72 x 20 um outlet channels • each side 108 x 4 um channels • separation bed 12.2 x 4.1 mm – 15,552 posts – 30 x 30 um
1937 A.Tiselius flow-counterbalanced electrophoresis Trans. Fraday Soc. 1937, 33, 524-531 1990 S.C.Lee et al EOF-counterbalanced CE 1994 C.T.Culbertson et al flow-counterbalanced CE
cyclic separation is not new… 1971 alternate pumping recycling LC: Biesenberger, J. A., Tan, M., Duvdevani, I., Maurer, T., J. Polym.Sci.B Polym.Lett.1971, 9 , 353–357.
cyclic separation is not new… 1993 synchronized cyclic CE: Burggraf, N., Manz, A., Effenhauser, C. S., Verpoorte, E., De Rooij, N. F., Widmer, H. M., HRC-J.High Resolut.Chromatogr.1993, 16 , 594–596.
vA vB detection window separation channel vsolution • in principle infinitely long separation column • small driving force: determined by circle circumference • resolution proportional to the square root of time Fourier transform chromatography
issues constant cross-section around cycle (pump?) high flow velocity for small channel diameters (pump?) overtaking of sample components (detection?) panic
shear flow pumping very high speeds possible nm gaps should lead to high N main problem: the end of the plate is the end of the pumping (no more than N=10,000 shown so far)
conclusions • cyclic separation can be done without sample loss • plate numbers and resolution increase with time • shear flow pumping most promising • deconvolution of detection signal by Fourier or wavelet transform
open questions • which signal deconvolution is best? • what dimensions are optimal for performance (theory)? • is there a simultaneous use for two independent stationary phases? • how about gradient elution?
Acknowledgment Oliver Hofmann Torsten Vilkner Xin Yang Eduardo Greaves, Prof. Jan Eijkel Sebastien Debesset Dirk Janasek Gareth Jenkins Joachim Franzke Qinetiq, UK Pfizer, UK Smiths Detection, UK Hitachi Ltd, Japan Universidad Simon Bolivar, Caracas, Venezuela Mercator professorship, Germany
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