New GCMS Applications

New GCMS Applications Analysis of Trace Fatty Acid Methyl Esters (FAME) in Jet Fuel, Sample Characterization by GCxGC/FID/MSD, Crude Oil Biomarkers Malgorzata Sierocinska Agilent Technologies Waldbronn 1

Introduction Increasing quantities of biodiesel and jet are being co-transported in multiproduct pipelines (MPP) In MPP transportation trace amounts of FAME can be found in jet parcels following biodiesel parcels due to FAME trail back. Following pipeline trials to establish the amount and profile of FAME trail back into jet fuel JIG PQ committee work on the effect of various FAMEs (up to 400 ppm) on the specification properties of jet fuel the main engine Commercial aircraft OEMs gave approval of limit of 5 mg/kg total FAME in jet fuel New method developed using single column GC/MS to detect individual FAMEs from 0.5 mg/kg to 50 mg/kg IP PM-DY/09 Method for Determination of FAME in Jet Fuel GC/MS with Selected Ion Monitoring 2

IP PM-DY/09 Method for Determination of FAME in Jet Fuel GC/MS with Selected Ion Monitoring 7890A GC Conditions: 5975C MSD Settings: Inlet: Electron ionization (EI) at 70 ev Temperature: 260 o C Source Temperature: 230 o C Mode: splitless Quad Temperature: 150 o C Sample size: 1 ul Scan Range: m/z 33 to m/z 320 SIM Groups: see next slide Column: HP-Innowax, 50m x 0.20 mm ID x 0.4 um Flow: 0.6 ml/min helium constant flow mode GC Oven: Initial temperature: 150 o C for 5 min. Ramp 1: 20 o C/min to 200 o C for 17 min. Ramp 2: 3 o C to 252 o C for 2 min. 3

Mass Spec SIM Ions Used for FAME Quantification FAME Species SIM Ions SIM Group Start Time Methyl Palmitate (C16:0) Methyl Heptadecanoate (C17:0)* 270 (mol. ion), 271, 239, 227 20 min. 284 (mol. ion), 253, 241 28 min. Methyl Stearate (C18:0) 298 (mol. Ion), 267, 255 32 min. Methyl Oleate (C18:1) Methyl Linoleate (C18:2) Methyl Linolenate (C18:3) 296 (mol. ion), 265, 264 35.5 min. 294 (mol. ion), 295, 264, 263, 262 36.5 min. 292 (mol. ion) 293, 263, 236 39 min. *C17:0 added to accommodate biodiesel made from animal fats 4

SIM/SCAN of Calibration Standard 0.5 mg/kg Each FAME in Dodecane Scan TIC C16:0 C17:0 C18:0 C18:1 C18:2 C18:3 SIM TIC 5

FAME Calibration 0 to 5 mg/kg Method uses this calibration for samples containing total FAME below 5 mg/kg 160000 140000 120000 100000 80000 60000 40000 20000 C16:0 R² = 0.9998 C18:0 R² = 0.9999 C17:0 R² = 0.9998 C18:1 R² = 0.9999 C18:2 R² = 0.9997 C18:3 R² = 0.9997 C17:0 C16:0 C18:0 C18:1 C18:2 C18:3 0 0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 6

Matrix Induced Retention Time Shifts of FAME Peaks Sample matrix can shift FAME peaks outside of their detection windows C16:0 +0.138 min. 50 mg/kg FAME Calibration Standard 50 mg/kg Each FAME Spiked in Jet Fuel C17:0 +0.098 min. C18:0 +0.083 min. C18:1 +0.076 min. C18:2 +0.067 min. C18:3 +0.054 min. 26 28 30 32 34 36 38 40 7

5 mg/kg and 1 mg/kg Total FAME Spiked in Jet Fuel Sample 1 Abundance 22000 Jet Fuel Blank 18000 14000 1 mg/kg Total FAME Spike 10000 C16:0 C17:0 C18:0 6000 2000 0 5 mg/kg Total FAME Spike C18:1 C18:2 C18:3 20 22 24 26 28 30 32 34 36 38 40 42 44 Min. 8

Qunatitative Results for 5 mg/kg and 1 mg/kg Total FAME Spiked in Jet Fuel Sample 1 5 mg/kg Total FAME Spike FAME Run 1 Run 2 Run 3 Std Dev C16:0 0.8 0.8 0.8 0.02 C17:0 0.9 0.8 0.8 0.01 C18:0 0.9 0.9 0.9 0.01 C18:1 0.8 0.8 0.8 0.01 C18:2 0.9 0.9 0.9 0.04 C18:3 0.9 0.9 0.9 0.02 Total 5.2 5.0 5.0 0.10 1 mg/kg Total FAME Spike FAME Run 1 Run 2 Run 3 Std Dev C16:0 0.3 0.5 0.5 0.09 C17:0 0.1 0.1 0.1 0.02 C18:0 0.1 0.1 0.1 0.02 C18:1 0.2 0.1 0.1 0.02 C18:2 0.2 0.2 0.2 0.01 C18:3 0.2 0.2 0.1 0.03 Total 1.0 1.2 1.2 0.12 9

Matrix Interference in Jet Fuel Sample 2 Interferences change depending on type of jet fuel 0.9 mg/kg Each FAME Spiked Second Jet Fuel Sample 0.5 mg/kg Std of Each FAME 20 22 24 26 28 30 32 34 36 38 40 42 44 Min. 10

Known Matrix Effects Raised C16:0 Detection Limit Data courtesy of Tom Lynch, BP Abundance Abundance 38000 36000 C16:0 C18:1 11000 10000 34000 32000 30000 28000 5 mg/kg 2 mg/kg 9000 8000 5 mg/kg 2 mg/kg 26000 24000 22000 20000 18000 1 mg/kg 0.5 mg/kg 7000 6000 5000 1 mg/kg 0.5 mg/kg 16000 14000 0.1 mg/kg 4000 0.1 mg/kg 12000 3000 10000 8000 2000 6000 4000 Reference fuel 1000 2000 0 Time--> 22.90 23.00 23.10 23.20 23.30 23.40 23.50 23.60 23.70 Time--> 35.10 35.20 35.30 35.40 35.50 35.60 35.70 35.80 35.90 36.00 36.10 36.20 36.30 36.40 36.50 Reference fuel 11

Dean s Heartcutting to Remove Matrix Interferences FID Restrictor S/S Inlet Aux EPC HP-Innowax MSD HP-5ms Capillary Flow Technology Deans Switch 12

MDGC Method:GC/MS Instrument Conditions Inlet: Temperature: 260 o C Mode: splitless Sample size: 1 ul Column 1: HP-Innowax, 30m x 0.25 mm ID x 0.5 um Flow: 1.0 ml/min He constant P (225 o C) Column 2: HP-5ms, 30m x 0.25mm ID x 0.25 um Flow: 2.0 ml/min He constant P (225 o C) CC Oven: Initial temperature: 150 o C for 5 min. Ramp 1: 20 o C/min to 200 o C for 17 min. Ramp 2: 3 o C to 252 o C for 2 min. MSD Settings: Electron ionization (EI) at 70 ev Source Temperature: 230 o C Quad Temperature: 150 o C Scan Range: m/z 33 to m/z 320 SIM Groups: see next slide Restrictor: 0.7m x 0.1 um ID deactivated fused silica 13

FID Signal Used to Set Heart-Cut Times Primary Column: HP-Innowax AVTUR Blank AVTUR 100 mg/kg Total FAME Spike FAME Primary Column Retention Time (min.) Heart-Cut Time (min.) C16:0 24.080 23.7 24.6 C17:0 29.151 28.9 29.5 C18:0 33.798 33.5 34.1 C18:1 34.841 34.5 35.1 C18:2 36.825 36.6 37.2 C18:3 39.570 39.3 39.9 Wider cut windows used to account for matrix induced retention time shifts 50 mg/kg Total FAME Std 22 24 26 28 30 32 34 36 38 40 14

Improve SIM Ion Groups for FAME Peaks Hydrocarbon mass peaks (mostly aromatics) in co-eluting jet fuel have little overlap with most FAME mass peaks Prototype Software recommends SIM ions to reduce background interference and improve S/N in the target peak elution time 51 77 167 115 190 141 Jet Fuel Blank Average Spectra 25.5 25.8 min. 210 230 258 304 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 74 43 143 227 270 185 239 5 ppm C16:0 Standard Average Spectra 25.5 25.8 min. 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 15

Expand SIM Ions Groups to Include FAME Base Peaks FAME Species SIM Ions SIM Group Start Time Methyl Palmitate (C16:0) Methyl Heptadecanoate (C17:0) 270(mol. ion), 271, 239, 227, 74(base) 284(mol. ion), 253, 241, 74(base) 20 min. 28 min. Methyl Stearate (C18:0) 298(mol. Ion), 267, 255, 74(base) 32 min. Methyl Oleate (C18:1) Methyl Linoleate (C18:2) Methyl Linolenate (C18:3) 296(mol. ion), 265, 264, 55(base) 294(mol. ion), 295, 264, 263, 262, 67(base) 292(mol. ion) 293, 263, 236, 79(base) 35.5 min. 36.5 min. 39 min. 16

HP-5ms Secondary Column Elution of FAMES After Heart-Cut HP-5ms Column MS Scan Data C16:0 C17:0 C18:0 C18:1 C18:2 C18:3 HP-5ms Column MS SIM Data Innowax Column FID Data C16:0 C17:0 C18:0 C18:1 C18:2 C18:3 17

Combination of Heart-Cutting MDGC and Base Peak SIM Ions 1 mg/kg total FAME Spiked in Jet Fuel Sample 2 (< 0.2 mg/kg each FAME) Improved detection of C16:0 FAME Better(?) detection of C17:0 C18:0, C18:2 and C18:3 Added matrix interference with C18:1 FAME C18:0 36.545 C16:0 27.399 C17:0 32.133 C18:1 37.336 C18:2 39.110 C18:3 41.558 26 28 30 32 34 36 38 40 Min. 18

Basic system layout for GCxGC FID/MSD Auto-sampler Switching valve s/s inlet PCM FID MS Tee 1 st column modulated 2 nd column Column 1 Column 2 Flow modulator MSD 19

Flow Modulator Diagram for Operation with the 5975C MSD Flow Modulation Interface for the MSD Second column MOD MSD 171mm x 110um restrictor Restrictor (0.4M x 0.25mm) FID MS Tee 20

GC Image processing of MSD GCxGC data TIC of heavy gasoline Spectra are library searchable 21

22 Kerosene: GCxGC with MSD

TIC: B20 Soy Biodiesel 23

C18:2 Mass Spectrum 24

5975C GCxGC Scan: 50-375 amu 19 scans/sec. (2.3 scans) Scan range Scans/sec Scans/peak 50-200 28 3.3 50-300 22 2.6 1. Naphthalene 2. Methyl naphtalenes 3. Dimethy naphthalenes 4. 1 methyl 4- phenyl methyl benzene 5. Anthracene 6. Methly phenanthrene 7. 9,10 dimethyl phenanthrene 8. n-c23 25

Paraffins and Olefins Mix 1 2 3 4 5 67 8 9 10 11 12 13 14 15 1. 1,8-Nonadiene 2. 1-Nonene 3. Nonane 4. 1,9 Decadiene 5. 1 Decene 6. 4 Decene 7. Decane 8. 3 Undecene 9. Undecane 10. 4 Decene 11. Dodecane 12. 2 Tridecene 13. Tridecane 14. 5 Tetradecene 15. Tetradecane 26

Paraffin and Aromatics Mix 12 13 14 15 16 17 18 19 20 21 22 1 2 3 4 5 6 7 8 9 10 11 1. Nonane 2. 3-methyl nonane 3. Decane 4. 3-methyl decane 5. Undecane 6. 3-methyl 1-undecane 7. Dodecane 8. 4-methyl-dodecane 9. 3-methyl-dodecane 10. Tridecane 11. Tetradecane 12. Butyl benzene 13. 1-methyl 4 propyl benzene 14. 1-methyl-4-(1-methylpropyl)-benzene 15. Pentylbenzene 16. 1-methyl butyl benzene 17. Hexyl benzene 18. 1,3 dimethyl butyl benzene 19. 1-methyl hexyl benzene 20. 1-methyl 2-n-hexyl benzene 21. 1-butylhexyl-benzene 22. 1-propyl heptyl-benzene 27

MSD as a Standard GC Detector 1.Easy setup wit autotune 2. Possibility of using e-methods 3. Sensitivity and positive confinfirmation 4. Possibility of creating RTL methods with associated libraries 5. Possibility of column backflush to keep the ion source clean 6. High reliability 7. Compact and easy to service 28

Biomarkers in Crude Oil Biomarkers are important in petroleum exploration for determining the age, biological source, and geological history of crude oils. They also allow characterization of crude oils in refineries and environmental monitoring. The characterization of crude oils for biomarkers is commonly performed by capillary GC in combination with HRMS or SIM. The application of SRM with a unique configuration of the GC allows for extended detection limits, higher throughput and higher analytical quality. 29

Experimental configuration 7890A GC Pressure / Flow Controller 7000A Injection Port Pulsed Splitless EI mode (70 ev) (300 C) Purged Ultimate Union 1.22 ml/min SRM mode Source 230 C 1.2 ml/min Column 1 Column 2 30

Analysis Speed 40m column 60m column 40 min 70 min Run times can be accelerated 30 minutes per cycle without loss in chromatographic resolution or substantial loss in signal by switching from a 60m column with He carrier gas to a 40m column with H2. The speed of the 7000A Triple Quadrupole mass spectrometer in SRM mode required only a change in dwell time from 50 to 20 msec to record the required 17 transitions with the same number of scans over the peaks. An experimental comparison with an uninterrupted 60m column (not shown) demonstrates that the use of the Pressure Controlled Tee configuration results in no degradation in chromatography. 31

Increased Throughput with Backflushing Backflush for faster turnaround, less carryover, and stable baselines Target compounds High boiling compounds seen in following solvent blank No Backflush Clean Solvent Blank With Backflush 32

Analytical Precision of MS-MS Reproducible biomarker concentrations in complex petroleum samples 33

34 Control Chart View

Linearity and Dynamic Range: Deconvolving Oil Mixtures A sophisticated understanding of petroleum systems requires the accurate deconvolution of oil mixtures derived from multiple source rocks. This problem is common where stacked source rocks exist in sedimentary basins. 35

36 Linearity and Dynamic Range: Deconvolving Oil Mixtures

Conclusions Advantages of using SRM over SIM identified thus far include increased sensitivity, better selectivity and the potential to greatly reduce analysis time Column backflush provided higher throughput with lower carry over Hydrogen and narrower bore columns reduced the run time nearly two-fold The scan speed, linearity, dynamic range and transition ratio stability of the triple quadrupole mass spectrometer allow the quantitative characterization and fingerprinting of petroleum samples and the deconvolution of petroleum mixtures. 37