MSL935004A Maintain instruments and equipment

Gas Chromatograph

Contents:
Quiz on last lesson

GC and its components -Review of basic Cleaning and maintenance of main components Trouble shootings Calculations involved in GC

Basic review of GC-8A Shimadzu

What are Applications of GC?

separation volatile and thermally stable organic compounds quantitative & qualitative

Important to know that:

Operates at high temperature usually 5-10 oC higher than highest bp of mix

Carrier Gas (Mobile phase):

Essential in moving analyte through column High Ultra Purity (less then 10ppm impurities)

Restricted choice Choice of N2 , Ar, He, Air, H2,

Storage Inert tubing

Comparison of gases
Gas Application Comments

He

General carrier or make-up gas

Excellent but expensive Can not use with ECD unless with make up gas
Cheap Not good for capillary as carrier gas (gives long run times)

N2

General make-up gas Carrier gas (packed column)

H2

Carrier gas (capillary columns) Combustion gas (FID, NPD & FPD)
Combustion gas (FID, NPD & FPD)

Highly explosive (oven trip sensor & careful venting) Best carrier gas (capillary)
Cheap and readily available

Air

Injector:
Via micro-syringe Sample not large for optimum column efficiency Slow Injection of large sample causes band broadening (loss of resolution)
Temperature should be about 50C higher than the boiling point of the least volatile component of the sample

Column (Stationary phase):

Packed -Inert solid supported material (glass or

stainless steel coil of 1-5m length and 5mm diameter) filled with stationary phase or coated with liquid stationary phase.
Carbowax- a brand name for polyethylene glycol

Capillarystationary phase is coated on inert solid tube

Common polymers use as stationary phases are polysilioxanes

Polar or Non-polar stationary phase

Temp must be controlled

Detector:
Detector Selectivity Detectability Flame ionization (FID) Thermal conductivity (TCD)

Most organic compounds Universal Halides, nitrates, nitriles, peroxides, anhydrides, organometallics Nitrogen, phosphorus Sulphur, phosphorus, tin, boron, arsenic, germanium, selenium, chromium Aliphatics, aromatics, ketones, esters, aldehydes, amines, heterocyclics, organosulphurs, some organometallics

100 pg 1 ng

Electron capture (ECD)

50 fg

Nitrogen-phosphorus

10 pg

Flame photometric (FPD)

100 pg

Photo-ionization (PID)

2 pg

Maintenance of GC
Preventive maintenance of your instruments will results in peak performance. Frequency of maintenance schedule will depends on how often you are using the instrument, if a GC is running multiple samples in an 8.0 hrs day, then you would be required to perform: Daily: check of leakage, performance of standards, run calibration curve and gas pressure. Weekly: check gas pressure cylinders, replace if necessary. Change septum, injector liner, perform manual leak test. Monthly: Check gas leak from primary gas supply to GC. Otherwise depending on how many samples are tested/day, how clean the samples are? Condition of syringe (auto sampler), environmental factors (temperature) will need to be considered to allow you in adopting a suitable schedule.

Septa High temp. septa are recommended to prevent injector and column contamination. Septum should be changed 30-50 manual injection and 75-100 for auto sampler injection. If septa leak, air will get inside column and it will rapidly damage the column. When changing septa, column temp. should be reduced to 35oC Sign of leakage or bleeding septa (ghost peak or poor reproducibility.

Injection Technique: Sample injection should be smooth and rapid, with quick removal of the syringe after injection, in order to avoid peak broadening. Injection volume is major issue, auto sampler is the solution. In manual injection try to rinsed and flushing the syringe 6 times.

Column Check for a sample problem by injecting a reference standard. If you get a good chromatogram, the problem most likely is sample related. If the chromatogram is not satisfactory, the problem probably is column or instrument related. Check for a column problem by replacing the column with a duplicate column, one known to provide good results under proper conditions. If results are good, the problem is related to the original column. If the symptom persists, the problem is related to the instrument. Incorrect installation of column is one source of leakage It is important to turn oven off and wait for 10-15 minute to cool, then turn carrier gas off (to prevent oxidation of column) before changing or installing a column.

A dirty detector creates noisy baselines. If the TCD is displaying problems such as a wandering baseline, increased noise level, or changes in response on a checkout chromatogram, it is probably contaminated with deposits from such things as column bleed or dirty samples. The TCD is cleaned by a process known as bakeout. Bakeout should be performed only after you have confirmed that the carrier gas and the flow system components are leak and contaminant free.

 must turn off the TCD and cap the detector column fitting to prevent irreparable damage to the filament caused by oxygen entering the detector. 1. Turn the detector off. 2. Remove the column from the detector and cap the detector column fitting. 3. Set the reference gas flow rate between 20 and 30 mL/min. Set the detector temperature to 400oC. 4. Allow thermal cleaning to continue for several hours. Then cool the system to normal operating temperatures.

Trouble shootings Please see GC resource on Moodle

The time between sample injection and an analyte peak reaching a detector at the end of the column is termed the retention time,

tR

What does tM represent?

Basic Definitions & Calculations

Term tm tr tr Capacity factor, k Definition/equation -Retention time of unrestrained peak -Retention time a component -Adjusted retention time tr = tr - tm =

( ideal

1-5)

Separation factor,

= k 2 / k 1

Term

Definition/equation

Resolution, R

Measure of separation between two peaks: 2(2 1) = 1 + 2

1 = 1 W2= 2

Term

Definition/equation
Peak Asymmetry factor, AF

= . 1
Where A = leading peak halfwidth B= tailing peak halfwidth

Number of theoretical plate, N

N = 5.45 (tr/W1/2)2

Height Equivalent to a Theoretical Plate,

where w1/2 is the peak width at halfheight HETP=L/N where L is the length of the column

Sensitivity: Slope of calibration plot (external stand)

LOD= 3

LOQ = 10

System Suitability Test The following criteria should be used to determine system suitability Injection of standard solution in 6 replicate Relative standard deviation in peak area and retention time should be evaluated RSD should be Peak Area (2%) and retention time (1%)