Running head: HANDLING TOXICOLOGY LAB SAMPLES 1

Handling Toxicology Lab Samples

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Appropriate sample collection and storage of biological evidence samples are important

to ensure correct analytical reviews. Further, correct handling of toxicology samples is crucial in

ensuring admissibility of the analytical results in court as valid depictions of the analytes being

investigated. The correct handling and analysis of toxicology blood samples start with the correct

documentation of the sample, the state of the packaging, and the integrity of all the seals applied.

Further, the chain of custody for the sample should be accurately recorded to ensure traceability

of the handling processes.

Note Taking

Sample handling is a crucial consideration during the pre-analytical phase. Unlike the

clinical setting where the time frame between sample collection and analysis is short, forensic

samples might potentially take longer to go through all the testing phases that are required.

Therefore, there are multiple measures that can be implemented to maintain the integrity of the

specimens after their collection. Among the key measures is the need for documentation and

description of the various parameters of the sample (Açikgöz, Hamamci, & Yildiz, 2018). The

blood specimen should be accompanied by an identifier label that highlights the case number, the

donor’s identifier information (such as their name, age, gender, and case number. These will be

important in the determination of the concentration of analytical chemicals to be applied in the

testing phase. Other information sets include the date and time of collection and the signature or

initials of the person collecting the sample undersigned with the date of collection. For the

provided blood sample with ethanol and cocaine, there is a need to document the type of sample

collected, the volume collected, the type of container that is used in the collection of the sample,

the potential suspect aspects of the sample (what the sample might potentially contain, in this

case being suspect for ethanol and cocaine) any presumptive tests that might have been carried
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out prior to the forwarding of the specimen to the lab. Further, the requisition for testing and any

additional special requests should be provided in the documentation accompanying the specimen.

Ideally, any improperly packaged or documented packages should be followed up early enough

or be returned to the submitting agency. Further, upon receipt of the blood sample specimen, the

receiving lab should inspect the specimen and appropriately document its condition and quantity

upon receipt.

Stability Issues and Storage Controls

Drug concentrations in toxicology blood samples can change during sample storage,

especially if the samples are stored at room temperature. Ethanol and cocaine are highly labile

drugs and present multiple stability issues that must be considered during storage. The

degradation of ethanol and cocaine in the stored toxicology samples might occur due to

hydrolysis, oxidation, or reduction (Cadamuro et al., 2019). Cocaine, for instance, contains

amide groups in its structure and is highly susceptible to breakdown by enzymatic and chemical

hydrolysis processes. Therefore, the addition of a preservative with a cholinesterase inhibitor

might be considered to decrease the breakdown rate for cocaine in the sample. The ester linkages

in the cocaine molecules are also implicated in the high lability rate for the drug in blood

samples. Ethanol concentration in toxicology samples is also subject to variation depending on

the storage parameters and considerations for the high volatility of the compound (Açikgöz,

Hamamci, & Yildiz, 2018). Ethanol is affected by evaporation, microorganism breakdown, and

oxyhemoglobin-mediated oxidation of the compound. In sealed sample containers, oxidation

remains one of the key contributors to the breakdown of ethanol. Therefore, the storage

parameters for blood samples suspected to contain ethanol and cocaine must consider,
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appropriate additives such as enzyme inhibitors, temperature ranges for storage, and filling levels

for the containers to prevent microbial contamination.

Blood samples need to be stored in cool and dry conditions with minimal exposure to

environmental elements of light, humidity, and air. These elements often lead to a higher

degradation rate for analytes in the sample. For short-term storage, refrigeration at 4 degrees

Celsius is advised (Majda et al., 2020). Long-term storage is not advised for blood samples

suspected to contain ethanol and cocaine. The two drugs are highly volatile and are labile to

degradation by multiple chemical and enzymatic pathways. Therefore, storage considerations

should be for short-term storage and analysis of the sample as soon as possible after collection.

The reactivity of the components of the vacutainer tubes also needs to be examined. For instance,

EDTA-laced tubes might react with metallic components of the blood and in turn, alter the

concentration of other analytes. Therefore, careful consideration for the preservatives should also

be made.

Analysis with Stability Considerations

The alcohol blood test for the toxicology report will determine the quantitative measure

of ethanol, its metabolites, and biomarkers in the blood sample. The blood alcohol content

measure will help to determine the potential for recent consumption of ethanol. Chronic use will

be determined through the use of other biomarkers in the blood such as carbohydrate-deficient

transferrin, which is indicative of the use of more than 50 to 80 grams of alcohol per day for two

to three weeks (Majda et al., 2020). Another biomarker that will be evaluated is phosphatidyl

ethanol, which closely relates to the amount of alcohol that one consumes. The stability

considerations for each test provide the time frame for which the specimen will still be viable.

For instance, the blood alcohol content can only be measured within six to 12 hours. The
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carbohydrate-deficient transferrin, on the other hand, is viable for up to three weeks. The test

processes for cocaine, on the other hand, will include gas chromatography/mass spectroscopy

processes to both detect and quantify the cocaine in the blood sample. Other metabolites that

result from the reaction between cocaine and ethanol can also be determined in the test

processes. For instance, cocaethylene is still detectable as a metabolite after a week.

Use of Preservatives

The use of a preservative is advised for blood samples containing ethanol and cocaine.

Ethanol neogenesis can occur during transportation and storage to affect the concentration in the

sample.t This process can be inhibited by sodium fluoride preservatives. Further, oxidation can

also occur in absence of enzyme inhibitors. Cocaine is broken down by esterase enzymes,

requiring the application of an inhibitor in the collection container. The stability of the sample

can also be improved by storage at 4 degrees Celsius.

The collection, documentation, and analysis of toxicology blood samples often need to

follow set standards for result validity and viability. Therefore, practitioners need to ensure that

there is effective labeling of the sample, document all activities related to the sample, and also

ensure that correct procedure are used in the chemical analysis phases. This way, they will be

able to get and report the correct results.

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References

Açikgöz, G., Hamamci, B., & Yildiz, A. (2018). Determination of ethanol in blood samples

using partial least square regression applied to surface enhanced raman

spectroscopy. Toxicological research, 34(2), 127-132.

Cadamuro, J., Lippi, G., von Meyer, A., Ibarz, M., van Dongen–Lases, E., Cornes, M., ... &

Simundic, A. M. (2019). European survey on preanalytical sample handling–Part 2: Practices of

European laboratories on monitoring and processing haemolytic, icteric and lipemic samples. On

behalf of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Working Group for the Preanalytical Phase (WG-PRE). Biochemia medica, 29(2), 334-345.

Majda, A., Mrochem, K., Wietecha-Posłuszny, R., Zapotoczny, S., & Zawadzki, M. (2020). Fast

and efficient analyses of the post-mortem human blood and bone marrow using DI-SPME/LC-

TOFMS method for forensic medicine purposes. Talanta, 209, 120533.