Thermo Scientific DFS

Transcription

1 Thermo Scientific DFS High Resolution Sector Field MS Instrument performance standards for Dioxin / Furan analysis Dirk Krumwiede, Product Specialist Trace MS

2 Overview: presentation main topics 1.) NEW TCDD Spec for DFS! 2.) Instrument Performance Standards 2

3 Thermo Scientific DFS High Resolution GC/MS DFS NEW Specification!! Behind the scenes

4 DFS: old TCDD Specification - 1 fg SN 8:1(4 ) D:\Xcalibur\...\SN346\tcdd1 9/3/29 4:43:44 PM RT: S/N 845:1 (4 ) E MS tcdd1 1 fg 2378-TCDD Relative Abundance Old SPEC Base Peak Retention Time : 1.5 Scan : 612 Intensity : 241 Calculated values Noise Range : minutes (65 scans) Baseline : 2.4 Signal To Noise : (4 sigma) Noise :.598 Algorithm: Statistical 4

5 21 DFS: New TCDD Specification - 2 fg SN 2:1(4 ) S/N 239:1 (4 ) fg 2378-TCDD Relative Abundance Made in Japan : DFS in Sendai, installed in March

6 21 DFS: New TCDD Specification 2 fg SN 2:1(4 ) 1 times repeated on DFS in Sendai, Japan D:\Xcalibur\...\tcdd_1 3/9/21 5:39:8 AM 2fg tcdd inj. volume: 1ul column:.1um oven: 12(2)-15-24()-6-28(1) Thermo TR-Dioxin 3m ID:.25mm film: RT: D:\Xcalibur\...\tcdd_1 3/9/21 1:48:35 AM 2fg tcdd inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) 1.6E3 RT: : : E2 MS tcdd_ MS tcdd_1 8 7 D:\Xcalibur\...\tcdd_4 3/9/21 2:54:44 AM 2fg tcdd D:\Xcalibur\...\tcdd_7 3/9/21 3:56:56 AM 2fg tcdd inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) 75 inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) 65 RT: RT: : : E E MS 85 MS tcdd_4 85 tcdd_7 5 4 D:\Xcalibur\...\tcdd_8 3/9/21 4:16:44 AM 2fg tcdd 8 D:\Xcalibur\...\tcdd_6 3/9/21 3:36: AM 2fg tcdd 8 inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) RT: : : 1 RT: E3 1.14E MS MS tcdd_8 tcdd_ D:\Xcalibur\...\tcdd_9 3/9/21 5:19:31 AM 5 2fg tcdd 8 D:\Xcalibur\...\tcdd_3 3/9/21 2:35:42 AM 2fg tcdd 15 inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) inj. volume: 1ul column: Thermo TR-Dioxin 3m 8.35 ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) RT: : RT: : E E Base Peak Calculated values Retention Time : 1.84 Noise Range : minutes MS (29 scans) MS Scan : Baseline : 19.3 tcdd_9 tcdd_3 Base Peak D:\Xcalibur\...\tcdd_5 Intensity : 162 Calculated values 3/9/21 3:15:13 AM 2fg tcdd Signal To Noise : (4 sigma) inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) 8 D:\Xcalibur\...\tcdd_2 3/9/21 2:13:49 AM 2fg tcdd Noise : 1.16 Algorithm: Statistical Retention Time : 1.78 Noise Range : minutes (279 scans) inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) Scan : RT: Baseline 242 : 19.3 : RT: Intensity : 734 Signal To Noise : (4 sigma) 1.83 : Noise :.84 Algorithm: Statistical E3 16E3 1.6E MS MS tcdd_5 tcdd_ Base Peak 5 8 Calculated values Base Peak Calculated values Retention Time : Retention Time : Noise Range : minutes (25 scans) 45 Noise Range : minutes (61 scans) 75 Scan : 133 Baseline : 19.6 Scan : 136 Baseline : Intensity : Signal To Noise : (4 sigma) 1.87 Intensity : 1214 Signal To Noise : (4 sigma) Noise : Algorithm: Statistical Noise : Algorithm: Statistical Relative Abundance Relative Abundance Relative Abundance Relative Abundance Relative Abundance Relative Abundance Relative Abundance Rel lative Abundance Relative Abundance Rela ative Abundance Base Peak Calculated values Base Peak Calculated values Retention Time : Noise Range : minutes (2 scans) Retention Time : Noise Range : minutes (132 scans) 45 Scan : 136 Baseline : 19.5 Scan : 136 Baseline : Intensity : 1244 Signal To Noise : 24.6 (4 sigma) Intensity : 1136 Signal To Noise : (4 sigma) Noise : Algorithm: Statistical Noise : Algorithm: Statistical Base Peak Calculated values Base Peak Calculated values 15 Retention Time : 1.85 Noise Range : minutes (152 scans) Retention Time : Noise Range : minutes (87 scans) Scan : 139 Baseline : 19.3 Scan : 128 Baseline : Intensity : 14 Signal To Noise : (4 sigma) Intensity : 1274 Signal To Noise : (4 sigma) Noise : Noise : Algorithm: Statistical Algorithm: Statistical # tcdd S/N (4 ) 1 2 fg 212 : fg 217 : fg 25 : fg 237 : fg 242 : fg 236 : fg 212 : fg 235 : fg 222 : fg 227 : 1 Base Peak Calculated values Base Peak Retention Time : 1.83 Noise Range : minutes (11 scans) Retention Time : 1.79 Scan : 133 Baseline : 19.5 Scan : 126 Intensity : 1444 Signal To Noise : (4 sigma) Intensity : 155 Noise : Algorithm: Statistical Calculated values Noise Range : minutes (129 scans) Baseline : 19.5 Signal To Noise : (4 sigma) Noise : Algorithm: Statistical 6

7 DFS: 2 fg TCDD Spec at IDÆA CSIC Barcelona D:\Xcalibur\...\1426_tcdd15 4/27/21 12:13:33 PM 1 ul 2 fg/ul 2378 TCDD (5 pg/ul 13C TCDD), new certified Std., 42 ev TR Dioxin 5ms 3 x.25 (.1 um film) 12 (2) () -6-29, R 1 K, 2 E6, 45 ev, GC links: 6 x.25 (.25).3 ml 8 C, ists 27 C, old 1 fg Std (seems OK), 42 ev RT: S/N 21:1 (4 ) 2 fg 2378-TCDD E MS 1426_tcd d15 Relative Abundance Base Peak Calculated values - 2 Retention fg Time : spec on 2 years old DFS, Noise Range : 7.64 two minutes (115 GCs scans) connected, running in routine Scan : 92 Baseline : 2.4 Intensity : 464 Signal To Noise : 2.4 (4 sigma) analysis (source 6 month in Noise DFS), :.553 Algorithm: only Statistical ion volume changed 7

8 DFS Sensitivity: New TCDD Specification 2 fg SN 2:1(4 ) fg 2378 TCDD S/N 2:1 (4 ) ative Abundance Rel

9 New spec on DFS Why? - High perfomance goals in DFS sensitivity - Customers ask for levels els << 1 fg (e.g. in Demos) - We want to challenge the competition 9

10 DFS in 26 in Beijing 2 fg native 2378-TCDD D:\Xcalibur\...\dk_tcdd22 9/26/26 11:49:18 AM man. 1 ul 2 fg tcdd (2) new syringe, BACK, DB5 3 m x.25 mm (.1), 12 (2)-15-24()-6-27,.8 ml/min, splitless 26 C, 1 min, aux 27 C, ists 27, 45 ev, ists 26 RT: Relative Abund dance fg native 2378-TCDD 1 / 5 dilution of certified 1 fg/ul Standard RT: AA: 83.5 RT: AA: 114 RT: AA: E dk_tcdd E dk_tcdd E dk_tcdd RT: AA: E fg 13 C labeled 2378-TCDD dk_tcdd22 1

11 New spec on DFS: Why 2 fg? - Evaluation - Evaluation of DFS testfield data 1 fg TCDD specs - Measurements of 2 1 fg TCDD standard on testfield DFS - Evaluation of 1 fg TCDD DFS installation specs - Measurements of 2 fg TCDD on > 1 testfield DFS Timeframe: about 1 year 11

12 Evaluation DFS: Instrument performance standard D:\Xcalibur\...\812_tcdd3 12/12/28 2:5:51 PM 1 ul 2-1 fg/ul 2378-tcdd (MXB, nonane), 4 ev, R 1 K TR Dioxin i 3m, ID.25,.1film, normal draw lens RT: E MS 812_tcdd3 Relative Abunda ance Base Peak Retention Time : Scan : 836 I t it 6891 Calculated values Noise Range : minutes (133 scans) Baseline : 18.6 Si l T N i (4 i ) Noise : Algorithm: Statistical Chromatogramm of one single injection! Intensity : 6891 Signal To Noise : (4 sigma) 12

13 New spec on DFS: Questions - Is there any technical improvement on DFS to achieve it? -Why not a 5 or 1 fg spec? 13

14 Thermo Scientific GC / HRMS: continued development D:\Xcalibur\...\lo_dioxin5 \ \l i 1/23/29 12:13:5813 PM 2 fg/ul 13C-2378-TCDD RT: Sector Field GC / HRMS MAT95 1 fg - 1: MAT95XL 1 fg - 4:1 55 Relative Abundanc ce DFS 4 1 fg - 8: DFS fg - 2: E MS lo_dioxin Base Peak Retention Time : 9.96 Scan : 92 Intensity : 639 Calculated values Noise Range : minutes (5 scans) Baseline : 9.2 Signal To Noise : (4 sigma) Noise :.465 Algorithm: Statistical 14

15 Comparison sensitivity DFS MAT95 1 fg tcdd (amount on column) on DFS and MAT95 Relativ ve Abundance DFS new ion optics > 3x signal intensity MAT95XP - conventional BE type instrument RT: AA: 9638 AH: 7491 RT: AA: 2911 AH: E F: MS ICIS dk_tcdd E F: MS ICIS mh1fg

16 Customer study confirms we are right CDC Talk and Poster at Dioxin 26 in Oslo DFS up to 5 times more sensitive than MAT95 on standards DFS up to 3 times more sensitive than MAT95 on real sample Quoted in our Dioxin/Furan application note 16

17 17 DFS Application Note on Dioxins

18 18 DFS Application Notes on Dioxins Citation of CDC study on comparision MAT95 - DFS

19 Thermo Scientific DFS High Resolution GC/MS Instrument performance standards

20 Instrument Performance Standards: 1.) Sensitivity standard One standard dwith ith6diff different native tcdd ddisomers at tdifferent concentrations ti (2 1 fg/ul) RT: Rela ative Abundanc ce Qualifier (ratio) mass trace 1379-TCDD 1368-TCDD 1378-TCDD 1478-TCDD 1234-TCDD 2378-TCDD 4.98E _tcdd Quantification mass trace 5 fg 1 fg 689E3 6.89E _tcdd3 25 fg 2 fg 5 fg 1 fg Solvent: nonane

21 Sensitivity Standard on DFS at IDÆA CSIC Barcelona RT: ve Abundance Relati unsmoothed data RT: AA: E RT: 1.54 AA: 181 RT: 1.82 AA: 435 RT: 1.91 AA: 853 RT: 11.1 AA: _tcd d21 Column: TR Dioxin 5 ms 3 m x.25 mm (.1 μm film)

22 Instrument Performance Standards: 2.) Reproducibility standard One standard with 6 different native tcdd isomers at same concentrations (5 fg/ul) TCDD 1378-TCDD 1234-TCDD 1379-TCDD 1478-TCDD 2378-TCDD Here: 5 times diluted Std. 2 (= 1 fg) Relative Abundance RT: 1.79 RT: 11.9 RT: RT: Solvent: nonane 22

23 Instrument performance standards (on DFS): set of two standards Standard 1 (2-1 fg/ul) and Standard 2 ( all congeners at same level) Relative Abunda AA: _tcdd11 RT: RT: 11.9 AA: 2953 RT: AA: AA: 597 RT: AA: E )S 1.) Sensitivity standard d 2.) Reproducibility standard _tcdd pg/ul 13 C labeled 2378-TCDD RT: AA: E _tcdd11 2 Relative Abund here: dilution all analytes at 1 fg/ul Original standard: all analytes at 5 pg/ul RT: 1.78 AA: AA: 913 AA: RT: AA: 15 RT: AA: _tcdd15 3 D:\Xcalibur\...\812_tcdd112 \ \812 12/1/28 1:18:2818 PM D:\Xcalibur\...\812_tcdd153\812 12/12/28 2:4:43 AM TR Dioxin 3m, ID.25,.1film, normal draw lens RT: TR Dioxin 3m, ID.25,.1film, normal draw lens RT: RT: 1.8 AA: E3 AA: 1166 RT: 11.1 RT: E3 1 1 AA: 9573 AA: 984 RT: fg/ul AA: RT: native tcdds AA: _tcdd11 812_tcdd15 4 RT: RT: RT: 11.9 RT: 1.79 AA: 2373 AA: AA: AA: 55 RT: 1.79 AA: E3 AA: 1273 RT: 11.9 RT: RT: E3 AA: AA: AA: ance1 RT: ance E _tcdd E _tcdd

24 Instrument Performance Standards: from CAMPRO Scientific 24

25 Instrument Performance Standards: 1.) Sensitivity standard What type of information can be obtained in just one injection? RT: 9,98-1,93 1 RT: 1,83 AA: ) Levels at 2/5 fg can be seen 2.) relative peakareas QM and RM 3.) Ratios at 6 different levels 4.) SN at 6 different levels 5.) Calibration curve (6 points) The current performance can be checked very efficiently Relative Abundance D:\Proben\...\2fg_2_1fg\tcdd_ :42:54 2fg 5fg 1fg 25fg 5fg 1fg tcdd inj. volume: 1ul column: Thermo TR-Dioxin 3m ID:.25mm film:.1um oven: 12(2)-15-24()-6-28(1) RT: 9,85-1,46 RT: 1,26 AA: RT: 1,12-1, ative Abundance Relative Abundance Rela RT: 1,2 RT: 1,26 AA: 187 AA: 347 RT: 1,25 AA: 386 RT: 1,2 AA: 187 RT: 1,7 AA: 3422 RT: 1,6 AA: 1866 RT: 1,52 688:1 AA: 742 RT: 1,25 AA: 386 RT: 1,2 AA: 279 RT: 1,7 AA: 4497 RT: 1,6 RT: 1,52 AA: 2135 AA: 795 9,85 9,9 9,95 1, 1,5 1,1 1,15 1,2 1,25 1,3 121:1 1,35 1,4 1,45 3,24E2 319, ,913 RT: 1,83 tcdd_12 AA: 7452 RT: 1,83 3,32E2AA: , ,911 tcdd_12 289:1 1,2 1,3 1,4 1,5 1,6 1,7 1,8 39:1 69:1 RT: 1,25 RT: 1,2 AA: 386 AA: 279 RT: 1,52 AA: 795 RT: 1,6 AA: 2135 RT: 1,7 AA: :1 1, 1,1 1,2 1,3 1,4 1,5 1,6 1,7 1,8 1,9 1 ul (2-1fg tcdd) measured on DFS in Sendai, Japan 25

26 Instrument performance standards Standard 2 1 injection for 6 times 1 fg/ul peaks, here QM mass traces RT: RT: 1.73 AA: 1281 RT: 1.79 AA: 1354 RT: 11.9 AA: 1263 RT: AA: 1361 AA: E _tcdd137 1.E3 Base Peak _tcdd33 1.E3 Base Peak _tcdd33 1.E3 Base Peak _tcdd33 Injection 1 26

27 Instrument performance standards Standard 2 1 injection for 6 times 1 fg/ul peaks, here QM mass traces RT: RT: 1.73 AA: 1281 R T: 1.74 AA: 1246 RT: 1.79 AA: 1354 RT: 1.8 AA: 144 RT: 11.9 AA: 1263 RT: 11.9 AA: 198 RT: AA: 117 RT: AA: 1361 R T: AA: 129 AA: 1242 AA: E _tcdd137 1.E _tcdd138 1.E3 Base Peak _tcdd33 1.E3 Base Peak _tcdd33 Injection 1 Injection 2 27

28 Instrument performance standards Standard 2 1 injection for 6 times 1 fg/ul peaks, here QM mass traces RT: RT: 1.73 AA: 1281 R T: 1.74 AA: 1246 RT: 1.79 AA: 1354 RT: 1.8 AA: 144 RT: 11.9 AA: 1263 RT: 11.9 AA: 198 RT: AA: 117 RT: AA: 1361 R T: AA: 129 RT: 1.74 RT: 1.79 RT: 11.9 RT: AA: 166 AA: 138 RT: AA: 1282 AA: 1275 AA: 129 AA: 1242 AA: 123 AA: E _tcdd137 1.E _tcdd138 1.E _tcdd139 1.E3 Base Peak _tcdd33 Injection 1 Injection 2 Injection 3 28

29 Instrument performance standards Standard 2 1 injection for 6 times 1 fg/ul peaks, here QM mass traces RT: RT: 1.73 AA: 1281 RT: 1.74 AA: 1246 RT: 1.79 AA: 1354 RT: 1.8 AA: 144 RT: 11.9 AA: 1263 RT: 11.9 AA: 198 RT: AA: 117 RT: AA: 1361 RT: AA: 129 RT: 1.74 RT: 1.79 RT: 11.9 RT: AA: 166 AA: 138 RT: AA: 1282 AA: 1275 AA: 129 RT: 1.74 AA: 127 RT: 1.79 AA: 1261 RT: 11.9 AA: 1117 RT: AA: 119 RT: AA: 137 AA: 1242 AA: 123 AA: 1128 AA: E _tcdd137 1.E _tcdd138 1.E _tcdd139 1.E3 Injection 1 Injection 2 Injection Injection 4 812_tcdd14 29

30 Instrument performance standards Standard 2 A Matrix of 4 x 6 peaks with almost same response RT: RT: 1.73 AA: 1281 RT: 1.74 AA: 1246 RT: 1.79 AA: 1354 RT: 1.8 AA: 144 RT: 11.9 AA: 1263 RT: 11.9 AA: 198 RT: AA: 117 RT: AA: 1361 RT: AA: 129 RT: 1.74 RT: 1.79 RT: 11.9 RT: AA: 166 AA: 138 RT: AA: 1282 AA: 1275 AA: 129 RT: 1.74 AA: 127 RT: 1.79 AA: 1261 RT: 11.9 AA: 1117 RT: AA: 119 RT: AA: 137 AA: 1242 AA: 123 AA: 1128 AA: E _tcdd137 1.E _tcdd138 1.E _tcdd139 1.E _tcdd14 (one injection) RSD tcdd RSD tcdd RSD tcdd RSD tcdd RSD 1368-tcdd RSD 1379-tcdd RSD RSD RSD RSD 1378-tcdd 1478-tcdd 1234-tcdd 2378-tcdd Do horizontal RSDs correspond to vertical RSDs? 3

31 Instrument performance standards (1 fg/ul all on DFS) Standard 2 Fun with Excel: statistics - ratios Entry Identifier RM1 Ratio (A) 1 RM1 Ratio (A) 2 RM1 Ratio (A) 3 RM1 Ratio (A) 4 avg stddev RSD (ver) 1368-tcdd tcdd tcdd tcdd tcdd tcdd avg avg / stddev avg RSD of RSDs stddev RSD (hor)

32 DFS: left 1 fg/ul tcdd; right 2 fg/ul tcdd Visual statistics TM DK, QM mass traces only RT: RT: R T: RT: E3 AA: 1361 AA: 1281 RT: 11.9 AA: 1263 AA: _tcdd Abundance1 Relativ ve Relative Abu Relative Abundan Abundance1 Relative undance ce RT: 1.8 AA: 144 RT: RT: AA: 129 AA: 117 AA: 123 RT: 1.79 AA: 166 RT: 1.74 AA: RT: 11.9 AA: 138 RT: AA: 1275 RT: 11.9 AA: 1117 RT: AA: 137 AA: 1128 AA: E _tcdd13 8 1E3 1.E _tcdd14 Abundance1 Relativ ve Relative Abu Relative Abundan Abundance1 Relative undance nce RT: 1.79 AA: 263 RT: 11.9 RT: AA: 17 AA: 215 RT: 1.78 AA: 234 RT: AA: 186 R T: AA: 298 RT: AA: 21 RT: AA: 186 RT: 1.79 AA: 185 RT: RT: AA: 174 AA: 151 RT: 11.8 AA: 14 RT: 1.73 AA: 193 RT: AA: 227 RT: 11.9 AA: 122 R T: AA: E _tcdd E _tcdd E _tcdd E _tcdd12 3 2E2 2.E _tcdd

33 DFS: Instrument performanced Standard 2: 1 fg, 2 fg, 1 fg 4 injections on each MS, QM mass traces only RT: RT: RT: RT: RT: 1.73 RT: E3 1.7E3 AA: 1361 AA: 2293 AA: 1281 RT: 11.9 RT: 11.9 AA: 1263 AA: AA: 1951 AA: _tcdd13 812_tcdd RT: E3 RT: E3 AA: 144 RT: AA: 2364 RT: 11.9 RT: RT: AA: AA: 2163 AA: AA: 117 AA: AA: bundance1 6 bundance tcdd _tcdd13 _cdd bundance Relati Relative Ab Relative Abunda ve Abundance1 2 nce Relative Abundance1 RT: 1.79 AA: 166 RT: 1.74 AA: RT: 11.9 AA: 138 RT: 11.9 AA: 1117 RT: AA: 1275 RT: AA: 137 AA: 1128 AA: Relat Relative Ab tive Abundance1 2 Relative Abunda 2 1.E _tcdd Relative Abundance1 Relat tive Abundance1 RT: 1.8 AA: 2168 RT: 11.9 AA: 274 RT: AA: 2148 AA: E _tcdd14 8 Relative Ab Relative Abunda Relative Abundance1 RT: 1.79 AA: 1273 RT: 11.9 RT: RT: AA: AA: AA: 1177 RT: 1.79 AA: RT: 11.9 AA: RT: AA: 1715 AA: 1784 RT: 1.79 AA: RT: 11.9 RT: AA: AA: AA: RT: RT: AA: 1115 AA: 1959 RT: 1.88 AA: AA: 1743 RT: AA: RT: AA: 6.6 RT: AA: E _tcdd E _ tcdd E ance _tcdd E3 RT: 1.73 AA: 2276 RT: 11.9 RT: RT: AA: 226 AA: AA: ance _tcdd E _tcdd E _tcdd

34 Thermo Scientific DFS High Resolution GC/MS Sensitivity standard on routine 6 m column

35 Fast Sensitivity check on 6 m x.25 (.25 film) column RT: e TR Dioxin 5ms 6 m x.25 mm (.25 μm) RT: AA: 6516 GC params: 2.8E _RomeD FS_tcdd11 1 ml/min He (const. Flow) 14 C (2 min) 3 C/min - 25 C - 3 C/min 31 C Rela ative Abundanc RT: 14. AA: 299 RT: AA: 67 RT: AA: 1569 RT: 14.9 AA:

36 Comparison 3 m (.1 film) to 6 m (.25 film) RT: RT: E3 Same height scaling RT: AA: 925 Relative Abundance RT: 1.15 AA: 456 RT: 1.42 AA: 694 RT: 1.5 AA: 1995 RT: 1.59 AA: _Rom edfs_tcdd25 Instrument had quite high baseline noise in this occasion 6.E Sensitivity standard for method 91127_Rom 85 edfs_tcdd21 optimization: 8 75 e.g. optimize injection volume for 7 65 required sensitivity level with given 6 conditions 55 / optimize conditions Relative Abundance RT: AA: 4549 RT: AA: RT: 27.2 AA: RT: RT: AA: 865 AA:

37 Sensitivity Standard on DFS at IDÆA CSIC Barcelona RT: Relative Abundance DB5ms 6 m x.25 mm (.25 μm film) Injection 1: 1 μl (GC start temp.: 15 C) 4 RT: AA: RT: 13.2 AA: RT: MA: 192 Injection 2: 2 μl (GC start temp.: 15 C) RT: AA: 13 RT: AA: 21 Injection 3: 4 μl (GC start temp.: 15 C) RT: AA: 23 RT: AA: 586 RT: AA: 285 RT: 13.8 MA: 667 RT: AA: 1489 RT: AA: 268 RT: 13.8 AA: 162 RT: AA: 157 RT: 14.2 AA: 2886 RT: AA: 5645 RT: AA: 2921 RT: 14.4 AA: 5674 RT: 14.4 AA: E MS 1428_6mt cdd12_ unsmoothed data Thermo Scientific Trace GC: Standard SSL injector 2.39E MS 1428_6mt cdd13 3 experiments: increasing injection volumes from 1 over 2 to 4 ul 4.6E MS ICIS 1428_6mt cdd16 37

38 Performance standards on DFS - Summary RT: Challenge level on 3 m column: good 9 perform. DFS ndance Relative Abu normal on 3 m column Challenge level on 6 m column 2fg 5 fg 1 RT: 1.73 RT: 1.8 even on DFS with bad performance 25 fg (on 3 and 6 m column) 1 fg RT: 11.9 RT: fg RT: fg RT:

39 Thank you for your attention! Questions / comments very welcome! 39