Introduction to Capillary GC
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1 Introduction to Capillary GC LC Columns and Consumables Simon Jones Chromatography Applications Engineer February 20, 2008 Page 1
2 Introduction to Capillary GC t r K c?? Kβ k = - tr t m? t m R s Page 2
3 Typical GC System Gas supply Injector Detector Data handling GAS Column Oven Page 3
4 CARRIER GAS Carries the solutes down the column Selection and velocity influences efficiency and retention time Page 4
5 VAN DEEMTER CURVES 1.00 N 2 H Small 0.25 Large He H u (cm/sec) Page 5
6 SAMPLE INJECTION Goals: Introduce sample into the column Reproducible No efficiency losses Representative of sample Page 6
7 SPLIT/SPLITLESS INJECTOR Carrier gas source Septum purge (~2ml/min) Ferrule Column Split vent Flow through injector = Column flow + Split Vent Flow Page 7
8 Typical Capillary Column Polyimide Coating Fused Silica Stationary Phase Expanded view of capillary tubing Page 8
9 DETECTORS Purpose: Responds to some property of the solutes Converts the interaction into a signal Immediate Predictable Page 9
10 DATA HANDLING Converts the detector signal into a chromatogram Integrator Software Program Page 10
11 COMPOUND REQUIREMENTS FOR GC Only 10-20% of all compounds are suitable for GC analysis The compounds must have: Sufficient volatility Thermal stability Page 11
12 SEPARATION PROCESS Page 12
13 TWO PHASES Mobile Phase Stationary Phase Solute molecules distribute into the two phases Page 13
14 DISTRIBUTION CONSTANT (K C ) Mobile Phase Stationary Phase K C = conc. of solute in stationary phase conc. of solute in mobile phase K C formerly written as K D Page 14
15 SOLUTE LOCATION In stationary phase = Not moving down the column In mobile phase = Moving down the column Page 15
16 SEPARATION PROCESS Movement Down the Column Mobile phase A A Stationary phase B 1 B 2 B B A 3 A 4 Page 16
17 KC AND RETENTION Fused Silica Tubing Stationary Phase Gas Flow Stationary Phase K c => Large retention K c => Small retention Page 17
18 KC AND PEAK WIDTH Time of Elution Fused Silica Tubing Gas Flow Stationary Phase Stationary Phase K c => Large K c => Small Page 18
19 THREE PARAMETERS THAT AFFECT KC Solute: different solubilities in a stationary phase Stationary phase: different solubilities of a solute Temperature: K C decreases as temperature increases Page 19
20 RETENTION TIME tr Time for a solute to travel through the column Page 20
21 ADJUSTED RETENTION TIME tr' Actual time the solute spends in the stationary phase tr' = tr - tm t r = retention time t m = retention time of a non-retained solute Page 21
22 ADJUSTED RETENTION TIME tr t r = tr - tm t r = t r = 3.16 min = time spent in stationary phase t m t r Page 22
23 TIME IN THE MOBILE PHASE All solutes spend the same amount of time in the mobile phase. Page 23
24 RETENTION FACTOR (k) Ratio of the time the solute spends in the stationary and mobile phases k = tr - t m t m t r = retention time t m = retention time of non-retained compound Formerly called partition ratio; k' Page 24
25 RETENTION FACTOR (k) Relative retention Linear Factors out carrier gas influence Page 25
26 PHASE RATIO (β) β = r 2d f r = radius (µm) d f = film thickness (µm) Page 26
27 DISTRIBUTION CONSTANT (Kc) K c = kβ k = t r t m β = r 2d f Page 27
28 RANGE OF RETENTION Average Fastest Slowest Time Page 28
29 PEAK SYMMETRY Symmetry = A B A B 10% height Tailing : Symmetry <1 Fronting : Symmetry >1 Page 29
30 PEAK WIDTH Peak width at half height Half height Peak width at base Page 30
31 PEAK WIDTH Page 31
32 EFFICIENCY Theoretical Plates (N) Large number implies a better column Often a measure of column quality Relationship between retention time and width Page 32
33 THEORETICAL PLATES (N) N = tr W h 2 t r = retention time W h = peak width at half height (time) Page 33
34 EFFICIENCY MEASUREMENT Cautions Actually, measurement of the GC system Condition dependent Use a peak with k>5 Page 34
35 ISOTHERMAL VS. TEMPERATURE PROGRAMMING Efficiency 9.31 N = 104, C isothermal N = 433, C at 5 /min DB-1, 30 m x 0.25 mm ID, 0.25 um He at 37 cm/sec C10, C11, C12 Page 35
36 SEPARATION VS. RESOLUTION Separation: time between peaks Resolution: time between the peaks while considering peak widths Page 36
37 SEPARATION FACTOR (α) α = k2 k1 co-elution: α = 1 k 2 = retention factor of 2nd peak k 1 = retention factor of 1st peak Page 37
38 RESOLUTION (Rs) R = 1.18 s t r2 - Wh1 + t r1 W h2 t r = retention time W h = peak width at half height (time) Page 38
39 RESOLUTION Baseline Resolution: Rs = W = h R = 0.84 % = 50 W = h R = 1.50 % = 100 W = h R = 2.40 % = 100 Page 39
40 Resolution N k α 1 R = s 4 k + 1 α N k α = Theoretical plates = Retention factor = Separation factor Page 40
41 INFLUENCING RESOLUTION Variables: N: column dimensions, carrier gas a: stationary phase, temperature k: stationary phase, temperature, column dimensions Page 41
42 Conclusions The GC is comprised of an inlet, column and detector that all work together to produce good chromatography Separation (via K c ) is based on 3 things: Solute: different solubilities/interaction in a given stationary phase Stationary phase: different solubilities/interaction of a solute (correct column selection is critical!) Temperature: K C decreases as temperature increases When in doubt, contact Agilent Technical Support! Page 42
43 Agilent J&W Scientific Technical Support (phone: US & Canada) * * Select option (fax) GC-Column-Support@agilent.com Page 43
44 Wrap-up E-Seminar Questions Thank you for attending Agilent e-seminars. Our e-seminar schedule is expanding every week. Please check our website frequently at: Or register for Stay current with e-notes to receive regular updates
45 Looking for more information on Agilent s GC Systems and Software? Agilent offers a full range of GC training courses including hands-on courses with the latest 7890 GC equipment as well as 6890 and 5890 courses. Each course includes a course manual for future reference and a certificate of completion. All courses are taught by industry experts. Call , Option 5 or visit to register today!
46 Upcoming GC and LC e-seminars Selection of a Capillary GC Column March 13, :00 p.m. EST Method Development March 18, :00 p.m. EST Installation, Care and Maintenance of Capillary GC Columns April 22, :00 p.m. EST
Introduction to Capillary GC. Page 1. Agilent Restricted February 2, 2011
?? Kβ? Page 1 Typical GC System Gas supply Injector Detector Data handling GAS Column Oven Page 2 CARRIER GAS Carries the solutes down the column Selection and velocity influences efficiency and retention
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