GC x GC Nueva dimensión en la búsqueda de desconocidos

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1 GC x GC Nueva dimensión en la búsqueda de desconocidos Marc Gibert CEO Ingeniería Analítica S.L. Su Partner Tecnológico en Integración de periféricos sobre Equipos Agilent

2 Soluciones para todos los niveles de exigencia Nivel V Solución completa con componentes tanto de Agilent como de otros fabricantes. Incluye HW,SW,Consumibles y materiales de referencia. Se garantiza las especificaciones de la normativa a la que se refiera. Field Solutions Nivel IV Incluye HW,SW,consumibles de Agilent probados y soportados por un VAR Se transfiere el conocimiento sobre la aplicación. Channel Partner Analyzer Nivel III Cada sistema sale de fábrica comprobado para cumplir el método regulado que se requiera Se desarrolla la documentación especifica requerida y se envía con cada solución. SP GC/MS Toxicology Screening Analyzer Analizador Agilent Nivel II Aplicaciones probadas con configuraciones concretas Comprobadas con múltiples muestras y bajo varias condiciones Se suministra todos los componentes requeridos(hw,sw,c&s,servicio) 3876AA LC/(Q)TOF Forensic/Tox PCDB and Spectrum Library kit G1734AA LC/QQQ Forensic/Tox Dynamic MRM Database Kit Kits aplicación Nivel I Notas de aplicación descritas de forma detallada y cuya fiabilidad ha sido probada. Notas de aplicación. Metódicas

3 AGENDA 1. Tecnología de Flujo Capilar 2. LTM (Low Thermal Mass) 3. Tecnología Agilent GCxGC 4. Modulador Fluídico Agilent 5. Estado del arte: Dual LTM GCxGC 5977 Ejemplos prácticos

4 Capillary Flow Technology - solves difficult application problems easily & opens up many new (and old) possibilities for GC & GC/MS

5 IF We Only Had A Technology That Provided Easy, Reliable Flow Structures In The GC Oven... It would open up many new (and old) capabilities for GC Column connections (connect pre-column) Change MSD columns (without venting) Backflush (Reverse flow through column) Detector splitter (effluent split to two or more detectors) Merge flows (2 columns to 1 MSD) Deans switch (heart cut select peaks to 2 nd column) Comprehensive 2-D GC (cut all peaks to 2 nd column) etc.

6 Types of Connectors Used In The GC Oven Advantages Limitations Metal Fittings Packed columns, reliable Not inert, no ferrule for capillary columns Press Fit Glass Low dead volume, inert, low cost Difficult to assemble, comes apart Graphite High temperature Sheds active graphite particles into sample path Polyimide Low initial leakage Loosens and leaks with oven cycling, solvent tailing

7 Challenges For Inside the Oven Devices Inertness (it is in the sample path) Low dead volume (it is in the separation path) Leak free (especially with repeated temp cycling) Fast thermal response (follow rapid oven ramping) High temp tolerance (GC oven can go over 350C) Reliable and easy to use

8 5 Key Developments in Capillary Flow Technology UltiMetal Ferrules Manifold Plates Agilent Ultimetal Plus Flexible Metal Ferrules. Easy to use, do not loosen or leak with oven cycling to 400 C Complex flow structures with low thermal mass Deactivation of Metal EPC Makes metal surfaces as inert as column Backflushing now possible, change MSD columns without venting, known column outlet pressure Calculators Accurately predict flows and pressures BEFORE installing devices

9 Capillary Flow Technology- Design a proprietary Agilent Technology Photolithographic chemical milling for low dead volume Diffusion bond two halves to form a single flow plate Small, thin profile provides fast thermal response Projection welded connections for leak tight fittings Deactivation of all internal surfaces for inertness

10 CFT Devices Provide Backflush Capability 2-Way Splitter with Makeup 3-Way Splitter with Makeup Deans Switch

11 3-Way Splitter With Makeup Effluent Splitter (3 Way) Det3 out Det2 out Aux EPC in Det1 out Column in

12 Capillary Flow Technology Restrictor 1 out to vent Column 1 In Restrictor 2 or Column 2 Nut Ferrule Channel Plate

13 Comparison of New Fitting with Polyimide Fitting Polyimide Fitting Exposure to polyimide and unpurged annular spaces is greatly reduced New Fitting Ferrule Ejector Hole

14 Fitting Design Minimizes Tailing Pentane test chromatogram Polyimide FID direct Capillary Flow fitting Capillary Flow Technology fittings avoid tailing with small but well swept dead volume

15 Capillary Flow Technology Devices Ultimate Union FID Ultimate Union Reliable precolumn connector Precolumn Column 7890 GC Gas Sampling Valve Tube to Connector Column FID Tube Connector Easy valve to capillary column connector Tube is 0.25 mm id and is deactivated 7890 GC

16 Splitters: Unpurged Tee Simultaneous detection with 2 detectors (but NOT MSD) Cannot do backflushing 1:1 split FPD:uECD Auto-sampler FPD P µecd 0.3 m X 0.25 mm id Effluent Splitter WITHOUT Makeup 7890A GC Column 30 m X 0.25 mm id X 0.25 um HP-5MS Det 1 OUT Column IN Det 2 OUT

17 Pesticides: Three Way Splitter with Makeup 1X method with 1:1:0.1 split FPD:MSD:ECD Auto-sampler Phosphorus FPD AUX EPC 3.8 psig uecd 3-Way Splitter with Makeup 7890 GC Column 30 m X 0.25 mm id X 0.25 um HP-5MS 5975C MSD

18 Dean Switch Heartcutting 2-D GC provides extremely high chromatographic resolution Cut Autosampler FID1 FID2 Deans Switch Column 1 Column A GC

19 2-D Separation of Sulfur Compound in Diesel Fuel Compound is completely resolved and can be analyzed with FID Diesel Fuel Heart Cut to Column 2 Column 1 - FID 1 Column 2 FID 2 Hydrocarbon Matrix Trace Sulfur Compound (4,6-DMDBT) min.

20 SPLITTER VARIABLE de Ingenieria Analitica para equipos Agilent flexibiliza el ajuste de respuestas en la multi-detección simultánea

21 Claves del SPLITTER VARIABLE: Emplea la fiable tecnología de AGILENT como base Control integrado en CHEMSTATION como parte de cada método Rango completo de SPLITT (0-100:100-0) Puede trabajar a RT constantes o a mínimo caudal Programable a lo largo del análisis Reproducibilidad y estabilidad del SPLITT a lo largo del análisis y entre diferentes análisis Adaptable a cualquier detector y método de trabajo Permite la realización de Backflush Permite el cambio de columna sin detener los detectores

22 SPLITTER VARIABLE DET #: FID, TCD, PFPD, SCD, NPD, MSD, AED, Sniffer... Auto-sampler AUX EPCs DET 1 SPLITTER VARIABLE Columna DET o 7890 GC

23 Capillary Flow Technology- Capabilities Solvent Bypass Heart Cutting (Deans Switch) Backflush Detector Splitting QuickSwap Modulation (GCXGC)

24 Summary Capillary Flow Technology solves difficult application problems easily. It opens up many new (and old) possibilities for GC and GC/MS systems.

25 Agilent LTM (Low Thermal Mass) Rapid Heating/Cooling System for GC, GC/MS

26 LTM Dual y microfluídica última generación Modulador CFT Divisor

27 LTM (Low Thermal Mass) Technology Direct heating of fused silica GC columns

28 LTM Cooldown Times (Standard Size) Temp (oc) Time (sec) 2m LTM 5m LTM 10m LTM 7890 GC Additional Cooldown times available on website for LTM column modules up to 30m

29 Interfacing LTM GC to an Agilent Gas Chromatograph Modules outside isothermal GC oven for fast heating/cooling Use same GC injectors, detectors, autosamplers, software, Independent and simultaneous temperature programming of: 1-2 col modules (standard, fast cool) 1-4 col modules (small) LTM Column Modules (standard 5 LTM module shown) LTM Integrated Software control Note: Front thermal shield removed to show LTM column modules Entire thermal shield should always be in place during operation

30 Fast Simulated Distillation Under 3 Minutes Using LTM 6x faster compared to conventional ASTM D2887 procedure pa pa C5 C6 C C8 C 9 C10 C11 C12 C14 C15 C16 C17 C18 C C24 ASTM D2887 Reference Oil LTM Module : 45ºC to 350ºC at 150ºC/min Faster than conventional GC ( o C/min max) C28 C32 C36 C40 min Calibration sample C5-C44 LTM Module : 45ºC to 350ºC at 150ºC/min min

31 pa Retention time repeatability similar to conventional GC Overlay of 10 runs Cracked gas oil: 45ºC to 350ºC at 150oC/min min General retention time repeatability data [by Agilent/Roger Firor] at LTM website Technology section or FAQ

32 PAH by LTM/GC/MS Fast Analysis - LTM 20 m x 0.18 mm ID x 0.18 µm DB-5MS Time--> Reference method Standard Oven 30 m x 0.25 mm ID x 0.25 µm DB-5MS Time-->

33 PAH by LTM/GC/MS Column contamination from just one prior run Contamination free when using backflush Time (min)

34 LTM Also Greatly Reduces Cool-Down Times, 9x Faster Heating/Cooling Cycle Times GC Run Time Cool Down 40 min min 20 min min 3 min + 2 min LTM/7890 9x Faster Cycle Time (200 o C/min)

35 Food/Flavor/Fragrance and Pharmaceutical Chiral Separations Chiral GC is most effective at low temperatures, yet complex samples usually require a broad temperature program to separate target compounds from background and to elute highly retained sample components. LTM/GC/MS uses a traditional column in the GC oven to separate target compounds, then heart-cut of target compounds to a chiral column in LTM module at a lower temp. optimal chiral separations are achieved while decreasing analysis cycle times and increasing chiral column lifetime EN Article Independent Temperature Control Using an LTM Oven Module for Improved Multi-dimensional Separation of Chiral Columns - 2D LTM/GC/MS, simultaneous/independent temperature programming of GC and LTM column module(s) Multidimensional Gas Chromatography with Capillary Flow Technology and LTM-GC, J Luong (et. al) Dow Chemical,R Mustacich Published:Journal of Separation Science, ISSN , Volume 31, pp Examples 2D LTM/CFT/GC using Deans Switch and LTM s simultaneous/independent temperature programming

36 Response Heart-cut Lavender Oil 0 Time Response Time Heart-cut Perfume

37 Tecnología Agilent GCxGC Sistema GCxGC LTM Dual Basado en el nuevo MSD 5977

38 GC x GC background information Why is the Application Important? - Real-world samples can be too complex for sufficient separation on one chromatographic phase - Target analysis is very complex samples - Excellent visualization of the sample - Powerful technique for hydrocarbon class determination Issues with current solutions - Non-integrated hardware - Lack of good data reduction software - Many based on thermal modulation requiring large quantities of cryo

39 Áreas de Aplicación GCxGC Comprehensive Chromatography es una herramienta muy potente para análisis cuantitativos y qualitativos de muestras con matrices muy complejas Petroquímica Forénsica Aromas y fragancias Medio Ambiente Muestras Clínicas Muestras Biológicas Donde la separación es muy dificil devido a las matrices muy complejas

40 7890B basic configuration The configuration is simple to get started in flow modulated GCxGC Split/splitless inlet, hydrogen carrier gas PCM module Modified TCD board Three way modulation valve Capillary Flow Technology modulator device Two columns: 30m x 0.25mm non-polar, 5m x 0.25mm polar FID at 200 Hz 7683 Auto Injector Other Possible Detectors : SCD, NCD, MSD,.. Data processing CG Image, LLC, Lincoln NE Zoex Corporation, Pasadena, TX 77505

41 Comprehensive 2-D GC (GCxGC) basics Consists of four parts: 1. A primary column (conventional separation) 2. A modulator 3. A second column (very fast separation) 4. Fast detector The peak capacity of the system is the product of the peak capacities of the two columns: result a lot of separation power The modulator does two jobs: 1. It collects effluent from the primary column 2. It transfers the collected effluent (in whole) to the secondary column This process is repeated approximately every 1.5 seconds, synchronized with the start of data acquisition GC x GC Chromatogram

42 Basic system layout 7683 Auto -sampler Switching valve PCM FID 1 st column modulated 2 nd column s/s inlet Column 1 Column A GC Flow modulator

43 Peak capacity ALS Conventional GC n= 1000 Column 1 GC x GC peak capacity is: 1000 x 25 = in this example Modulator n = 25 Column 2 Fast GC Detector

44 Agilent s flow modulator : Differential Flow Using Design by John V. Seeley, Oakland University Flow modulator eliminates the need for cryo. Sample compression controlled by flow ratios occurs in the collection loop and is quickly injected into the second column, resulting in very narrow and tall peaks.

45 Load or collect step 1 ml/min Split/Splitless Inlet H2 Modulation Valve Collection channel Column 1 (25 30 M) Load time must not be longer than time to fill collection channel Collect Flow direction Column 2 (5M) FID Flow Modulator 20 ml/min

46 Inject step 1 ml/min Split/Splitless Inlet Inject Flow direction H2 Modulation Valve 20 ml/min Collection channel Column 1 (25 30 M) Collection channel is quickly injected into second column in about 0.1 seconds Column 2 (5M) FID Flow Modulator 20 ml/min

47 Chromatographic parameters Agilent 7890A Inlet: Split/splitless Detector: FID Data rate: 200 hz Other pneumatics: PCM Auto injector: 7683 Syringe: 5 ul Carrier: Hydrogen Inlet Pressure: 21.5 at 50 o C, constant flow PCM: 20 ml/min, constant flow Oven Program: 50 o C (1.0min) to 260 o C (4 5 or 8 o C/min.

48 Typical modulation parameters Modulation start: Synchronized with start of data acquisition Modulation Valve on: 0.12 seconds typical (Inject) Modulation Valve off: 1.40 seconds typical (load) Collection channel: 15 cm x 0.45 mm ID Three-way valve: HP , 12VDC

49 N-butylbenzene unmodulated modulated

50 Kerosene raw data Zoom from 15.8 to 16 min Alkane, mono-aromatic, and di-aromatic separated in 1.5 seconds

51 Heavy gasoline raw 2D data Note hydrocarbons being separated in each 1.5 second modulation

52 GC X GC modulation basics 1.Acquisition D-GC chromatogram (at the end of the 1 st column in blue) modulations shown Coelution of 3 compounds! 2. Transformation Raw 2D-GC chromatograms (at the end of the 2 nd column) Take slices of the co-eluting peaks and inject quickly to another column of different selectivity red peak elutes last now and green peak elutes first and all 3 completely separated!

53 2 nd Dimension (fast GC) 2D Image 2 nd Dimension chromatograms stacked Reconstitute the peaks by combining each of them from each chromatogram 1 st Dimension Chromatograms produced by 8 modulation cycles

54 Flow modulation: Detergent perfume base:7890a 2D GC is ideal for application of pattern recognition techniques to determine product quality, authenticity, adulteration, etc. Small degree of wrap around on this compound

55 Flow modulation: (GC x GC) of diesel fuel: 7890A Naphthalene p-xylene o-xylene Toluene C9 Methyl-naphthalenes C12 di-aromatics mono-aromatics Alkanes C16 GC x GC Image: Showing the normal B.P. distribution (1 st dimension) Also shows the hydrocarbon class clusters Consistent RT for alkanes in 1 st dimension showing precise modulation Comparable peak in 2 nd dimension band shows minimum peak broadening with flow modulation

56 Flow modulation: (GC x GC) of Kerosene: 7890A Di-aromatics Mono-aromatics Alkanes Modulation 1.40 second collect 0.11 second inject Column 1: HP-5MS 30m x 0.25mm x 0.25um Column 2: INNOWAX 5m x 0.25mm x 0.15um Oven rate: 8 o C/min

57 Summary for Agilent flow modulation GC x GC Reliable Setup: Based on capillary-flow- technology, easy to setup, high performance chromatography, and reliable. High temperature operation: Operation to 350 C expands applications No Cryogen Required: Flow modulation means no tanks of Liquid N 2 or CO A Enabled GC x GC: Capillary- flow-technology ready, synchronized periodic events ensure precise modulation, control from modified TCD board Comparable resolution without N2: Cap Flow Technology allows low dead volume and precise flow control, resulting in minimum peak broadening even without cryo-focusing. Peak widths on the second column are typically 60 to 100 ms at half maximum. Sensitivity: Approaches that obtained by thermally modulated systems

58 Principio Básico de Operación Modulador microfluídico Agilent Partes 1. Columna primaria (Separación convencional) 2. A modulador 3. Columna secundaria (separación muy rápida) 4. Detector rápido (MSD5977, FID, SCD )

59 Único horno: única rampa de temperatura

60 Doble horno: doble rampa de temperatura para 2D Ajuste de la resolución de la segunda dimensión

61 Doble horno: doble rampa de temperatura para 2D Speed-UP en LTM

62

63 FID Cromatograma gradiente normal (>60 min)

64 FID Cromatograma a doble velocidad (<30 min)

65 Coeluciones en 1D Zona min

66 Componentes resueltos

67 Comparación High Speed Normal Speed - FID

68 Comparación High Speed Normal Speed - MSD

69 Coeluciones en 1D Zona min

70 Componentes resueltos

71 Comparación High Speed Normal Speed - FID

72 Comparación High Speed Normal Speed - MSD

73 Investigación de superposición de compuestos 98% corr Ref vértice 96% corr

74 Identificación univoca

75 +1200 componentes en plantilla

76 7 componentes vs +115 posibilidades en la zona

79 Inercia del sistema para compuestos muy activos 8 ppm Azufre totales SCD

80 Separation of 4,6-Dimethyldibenzothiophene in diesel 4-Ethyl,6-methyldibenzothiophene 4,6-Dimethydibenzothiophene Separate run with just the dibenzothiophenes

81 ... la pregunta es clara... Sigue queriendo ver cine mudo y en blanco y negro? O.. prefiere la tecnología actual?

82 Gracias por su atencion Marc Gibert CEO Ingeniería Analítica S.L. Tel