Supporting Accurate Assessments of Nicotine Exposure

By Mira Doig, ABS Technical Sales Lead

woman smoking a cigarette

Throughout the years, ACM Global Laboratories via its bioanalytical lab, ABS, has developed a dedication to the bioanalysis of nicotine and its metabolites to aid smoking cessation, via a number of extensive research studies. 

These studies include:

  • Supporting GSK’s comparative bioavailability studies exploring patches and other novel formulations of nicotine, such as gums, lozenges and sprays
  • Supporting Niconovum to develop their Zonnic range of nicotine replacement products 
  • Assay support to BAT & Swedish Match to explore novel formulations of tobacco SNUS and SNUS replacement products
  • Most recently it includes helping a host of E-Cig manufacturers to develop efficient delivery devices

Innovation in the Field of Nicotine Analysis

As part of ABS’s continued experience in the field of nicotine research, the team has a validated LC-MS/MS assay for nicotine analysis in plasma, which has proven to be difficult to achieve in most labs, and has perfected the process by implementing a number of failproof measures to ensure a proper LLOQ of 0.1 ng/mL. 

These measures include the following requirements: 

  • All staff in the building are to be non-smokers (and if they do smoke, they will lose their job)
  • No visitors are allowed in the labs if they are smokers especially if we are performing nicotine assays
  • The lab maintains a positive air pressure, so if there is any nicotine in the office area, it does not go into the lab area
  • All reagents, solvents and consumables are screened for nicotine contamination prior to use

Better Testing for Treating Nicotine Addiction

Contributing to the accurate assessment of nicotine exposure would be the team’s assay development for cotinine. The measurement of this stable major metabolite of nicotine with a half-life of 20 hours compared to nicotine’s 4, in a variety of matrices such as saliva, plasma and urine, helps to determine nicotine dependence, is the optimal measure for smoking cessation assessment and can be used to evaluate passive smoking levels. In addition, ABS has developed other assays that can be used. ACM/ABS has the ability to measure an individual’s nicotine metabolism rate which can aid in prescribing the best medication for treating nicotine addiction. We can also measure anabasine and 3-HPMA to differentiate between NRT and tobacco usage. 

nicotine assays infographic

Other Assays Used to Assess Smoking

There are other assays available to assess smoking, such as measuring carbon monoxide (CO) with a breath test. It is inexpensive, non-invasive and good for assessing heavy smokers, however, it’s not quantitative, cannot be used for smokeless tobacco or NRT, has a short half-life, and there is a high percentage of false results. 

Antibodies raised against cotinine (monoclonal antibody (mAbs) have also been utilized to assess nicotine usage via a urine dipstick which isn’t sensitive or quantitative but can be used at point of care. Unfortunately, like CO measurement, there are often many false negatives and it tends to only pick up regular heavy smokers. In comparison, using the mAbs in ELISA (enzyme linked immunoassays) assays can generate quantitative results, but samples must be sent away and are not available at point of care. Additionally, these assays are expensive and they have poor sensitivity. It has also been found that different kits can produce non-comparative results. 

Saliva and Plasma Testing for Nicotine Analysis

The ABS team has found that using saliva or plasma, collected mid to late afternoon for best results, are best suited for testing cotinine utilizing Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). The LC-MS/MS assay delivers results suitable for the assessment of nicotine dependence in smoked and smokeless forms, in addition to the ability of measuring environmental exposures. Unlike ELISAs, results from different labs are comparable. 

Although the equipment is expensive, and chromatography is sophisticated, the advantages of nicotine and cotinine assays outweigh any disadvantage. Analysis times average five minutes per sample. LC-MS/MS provides the ultimate specificity and has the ability to assay other analytes in the same assay. Additionally, automation allows for optimal cost saving for large batches of samples. After samples have been measured, ABS can also provide an interpretation of the results.

Qualitative vs. Quantitative Data in Nicotine Research

Interestingly, the majority of clinical studies forming the basis of the NICE and US Public Health Guidelines smoking cessation guidance use cotinine assessments. However, using cotinine measurements remains unmentioned within these cessation guidelines. Seemingly the belief is that asking the patient if they have stopped is sufficient. Given ABS’s experience supporting patient surveys, verbal answers often contradict actual cotinine measurements. How, if not with the contribution of a cotinine assay, can nicotine researchers supply accurate data or smoking cessation practitioners accurately support their patients? 

woman breaking a cigarette

Choose the Right Partner for Bioanalytical Research

At ABS, we believe cotinine assessment is key for successful smoking cessation, and the ABS bioanalysis team is prepared to take on your nicotine research project, utilizing only the best technology and best practices. For more information, visit our Bioanalytical Services.

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