Experiment 8

Identifying Lidocaine in a Urine Sample by Gas Chromatography

Jessica Nicholson and Madi Kreitz

11/8/2017

Mike Miller
Introduction:

This experiment focuses on the detection of drugs of abuse from urine samples. The

process of liquid-liquid extraction will be utilized as a method to isolate the drug lidocaine, and

detect its concentration from a 25.0 mL of the provided urine sample. Lidocaine is a

commercially available local anesthetic that is used for temporary numbing of the area where it

is injected. It has been illegally used on horses to improve their performances when injured in

races.

To figure out the concentration of lidocaine it must be isolated from urine due to the fact

that urine has many non-volatile compounds that cannot be analyzed directly by gas

chromatography (GC). This is extracted by the use of a liquid-liquid extraction which is a

common organic chemistry method of separation. Lidocaine is extracted with the addition of

sodium bicarbonate and then a dissolving agent for the lidocaine to dissolve in the organic layer.

These techniques are used in the every day lives of forensic scientists in things like drug

tests, or to see if people who have passed had a abnormally high amount of drugs within their

systems. This process is important because it shows exactly how this will be utilized.

Experimental Procedure:

1. Obtain a 50 mL volumetric flask and add 1.00 g or lidocaine

2. Fill the flask to the mark with dichloromethane to get a 20 mg/mL lidocaine solution

(Stock solution A)

3. Take 5.0 mL of this stock solution A into a 10.0 mL volumetric flask and then add

enough dichloromethane to fill to the mark to get a concentration of 10.0 mg/mL (stock

solution B)
 4. Take 5.0 mL of this stock solution B into a 10.0 mL volumetric flask and then add

enough dichloromethane to fill to the mark to get a concentration of 10.0 mg/mL (stock

solution C)

5. Take 5.0 mL of this stock solution C into a 10.0 mL volumetric flask and then add

enough dichloromethane to fill to the mark to get a concentration of 10.0 mg/mL (stock

solution D)

6. These solutions will be utilized later in the experiment

7. Under the hood, add 25.0 mL urine sample into a separatory funnel

8. Add 5% Sodium bicarbonate solution, small portions at a time until the pH indicates

basic and precipitation occurs

9. Add 25.0 mL of dichloromethane into the funnel to dissolve lidocaine

10. Vent and shake the funnel several times to release any built-up gas

11. Drain out the organic layer and then dry it with sodium sulfate

12. Prepare GC sample by adding 2 mL of the dried lidocaine into a GC sample vial

13. Run all GC analyses for results

14. Construct the calibration curve on excel

Chemical Hazards:

 Gloves, goggles, and lab coats should be worn at all times

 Do not consume any of the experimental materials that are used in this lab

 Lidocaine may be toxic to the cardiovascular system, central nervous system

 Methanol is poisonous and flammable, do not make contact with skin or mouth
Results:

Solution Concentration (μg/mL) Peak Area Retention Time (Min)

1 2.5 740.7 4.81

2 5.0 1574.4 4.82

3 10.0 3503.8 4.84

4 20.0 7218.5 4.88

5 Unknown 2721.0 4.84

Table 1

Table 2. Concentration vs Peak Area

Calculations:

𝑦 = 372.35𝑥 − 231.4 Table 3. Calculation of Unknown Concentration

2721.01 = 372.35𝑥 − 231.4

𝑥 = 7.93 𝜇𝑔/𝑚𝐿
Conclusion:

Through this experiment, the lidocaine concentration in the unknown sample was found

to be 7.93 μg/mL. This concentration was found through the use of Gas Chromatography. A

range of concentrations from 2.5 μg/mL to 20.0 μg/mL were analyzed and their retention times

and peak areas were all found. As the concentration increased, it would increase by doubling

which in turn would make the peak area increase, which would provide a straight trendline graph

as shown in table 2. This trendline covers all the concentrations between the given range and it

relates them to their peak areas, or what they should be.

The unknown present in this experiment was now able to be analyzed by GC, from here

the retention time and peak area is found. It is possible for a calculation to be made using the

given graph equation in a calculation as seen in table 3. So, from this the concentration of

lidocaine in the unknown substance will be 7.93 μg/mL due to gas chromatography results.

Possible sources of contamination in this lab is the GC syringe not being fully cleaned or

rinsed between the cycles and analysis of different molecules. At the start of lab, the GC should

be loaded with acetone to clean and a waste vial to spit it out. Another error could be the uneven

or incorrect making of the sample concentrations that give the trendline, if one is too high than

all will follow in its footsteps. Last, if the lidocaine isn’t successfully and completely separated

from the unknown mixture it would have had a hard time reading on the GC machine. All of

these can be avoided by a careful procedure and process in the lab area.

Work Cited:

Huynch, L., Henck, C., Saxton, K., & Wang, J. (n.d.). Introduction to Forensic Chemistry:

Laboratory Manual (Vol. 1). University at Albany.