The Detection of Propofol Glucuronide in Human Hair Using LC-

MS/MS: Case Study and Survey of High-Risk Health Professionals

Background

Despite initiatives to increase awareness, alcohol Propofol (2,6-

diisopropylphenol) is a commonly used agent for general

anesthesia because of its short duration, quick recovery time, and

minimal side effects (1). Although not a controlled substance in the

United States, propofol abuse and misuse by medical professionals

has increased 5-fold between 1997 and 2011 (1). Propofol

detection in biological fluids has a very short detection window

and the presence of propofol in hair does not distinguish between

ingestion and environmental exposure. A metabolite, propofol

glucuronide, is formed in the liver and may be detected in the hair

(2).

Objective

The purpose of this study is to validate a method for the detection

of propofol glucuronide in hair and apply this method to a case

study and a survey of high-risk health professionals

Methods

• Sample Prep

• Wash hair

• Hexane

• Methylene Chloride

• Methanol

• Powder hair

• 3mL DI Water

• Sonicating water bath

• No heat

• Overnight

• Solid Phase Extraction

• Quaternary amine w Cl counter

ion

• UCT CUQAX12Z

• Condition Cartridge

• 2mL MeOH

• 2mL DI Water

• Load Sample onto cartridge

• Wash Cartridge

• 2mL DI Water

• 2mL MeOH

• Elute compounds

• 2mL MeOH w 2% Formic Acid

• Evaporate 40 oC under N2

• Reconstitute in 100 mL DI Water

Selected Validation Results

Conclusions

This method demonstrated sufficient precision, accuracy and

robustness with an analytical measurement range between 8

pg/mg and 100 pg/mg. While being sensitive enough to identify

two individuals in a survey of high-risk substance abuse treatment

participants, the method was not able to identify propofol

metabolite in the hair of a woman receiving a routine quantity of

propofol administered during a short surgical procedure. This

method provides substance abuse treatment professionals with

another objective tool to identify propofol abusers

Acknowledgements

This project was funded internally by USDTL.

J. Jones, M. Jones, C. Plate, D. LewisUnited States Drug Testing Laboratories, Des Plaines, Illinois USA

Performance Characteristics

Limit of Detection 4 pg/mg

Limit of Quantitation 8 pg/mg

Upper Limit of Linearity 100 pg/mg

Intra- and Inter-assay bias < 15%

Intra- and Inter-assay imprecision < 15%

3 Freeze-thaw cycles Stable

4 days room temp on autosampler Stable

• Liquid Chromatography

• Synergy Polar RP Column

• 50mm x 2mm x 2.5 mm

• 1200 Agilent HPLC

• A = 0.1% Formic acid

• B = ACN w 0.1% Formic Acid

• Flow = 0.25 mL/min

• Isocratic 32% B

• Injection volume 10 mL

• Mass Spectrometry

• AB Sciex 5500 Tandem MS

• Negative electrospray

• CUR = 30

• CAD = 5.0

• IS = -4500

• TEM = 500

• GS1 = 60

• GS2 = 40

• Dwell = 100 msec

• Propofol glucuronide

• m/z 353.1 => m/z 176.7, 112.6, 84.7

• Propofol glucuronide-d17

• m/z 370.1 => m/z 193.7, 369.7

0.02

0.10

0

0.04

0.08

0.12

1990-1997 1995-2005

10

-yr

inc

ide

nc

e (

%)

10-yr incidence of propofol abuse is up 5-fold

at academic anesthesiology programs (1)

Comparison with Other Findings

The assay was validated according to commonly accepted

guidelines. The assay proved to be stable and consistent

throughout the validation study.

A survey of 300 medical professionals that were participating in an

addiction treatment program were subjected to the method. Two

specimens contained detectable amounts of PPFG, with one just

below the LOQ at 7 pg/mg and one at 108 pg/mg. These findings

are compared to the findings of 7 suspected propofol abusers

reported by Kim et al (2) and charted above.

An authentic hair specimen from a 50 year old women anesthetized

for 15 minutes with propofol did not contain detectable PPFG

Results from Authentic Specimens

References:

(1) Wischmeyer, P. et al (2007). A survey of propofol abuse in academic anesthesia programs. Anesthesia &

Analgesia, 105(4), 1066-1071.

(2) Kim, H., Cheong, J., Lee., J., & In, M. (2013). Rapid and sensitive determination of propofol glucuronide in

hair by liquid chromatography and tandem mass spectrometry. Journal of Pharmaceutical and

Biomedical Analysis, 85, 33-39.