Leveraging Your NSF Certification: Now that you've received NSF Certification ... why not leverage the Mark to your advantage.
by Kerri LeVanseler, Ph.D., NSF International Technical Manager, Chemistry Laboratories
Doping in sports has secured several headlines in the last few years, with major U.S. publications such as Sports Illustrated and The Wall Street Journal questioning the integrity of nutritional supplements and other sports nutrition products and their potential for causing harm. Athletes must be ever vigilant when taking over-the-counter products to avoid consuming compounds that are prohibited by sports organizations. More recently, concerns have arisen about excessive amounts of stimulants in consumer products including both dietary supplements and food products. Parents and coaches are being warned about the dangers of sports nutrition products that young athletes consume in an attempt to improve their athletic performance.
The U.S. Food and Drug Administration has issued warnings about more than 70 weight-loss supplements that include potentially dangerous ingredients. Steroids, steroid-like ingredients, stimulants and prescription drugs found in contaminated products have been reported to cause serious side effects including liver damage, stroke and kidney failure.
With so much at stake, it is clear why the nutraceutical industry has responded with a renewed fervor for demonstrating the quality and safety of nutritional supplements. Third-party testing and certification of nutritional supplements and sports nutrition products has remained the champion of manufacturers that seek a reliable method for verifying the integrity and safety of their products. But as new designer drugs and steroid-mimicking compounds evolve, so must the technology used to test for these substances. Much of the success of screening for athletic banned substances lies on staying one step ahead of the culprits.
This means that pioneering laboratories, such as NSF International’s nutritional supplement testing lab in Ann Arbor, Michigan, are faced with the challenge of relentlessly developing new test methodologies using the most sophisticated instruments. This article discusses the latest technology for screening supplements and sports nutrition products for even the most complex banned substances.
The World Anti Doping Agency’s list of prohibited substances covers a veritable cornucopia of stimulants, narcotics, diuretics, beta-2 agonists and steroids. But even with so many performance-boosting compounds identified for testing, more are being developed. In some cases, dietary supplements are laced with untested or undetectable banned substances. Clever chemistry can be deceiving by: 1) creating new compounds that act the same as banned substances, hence mimicking the same performance boosting effect, and 2) masking banned substances by making them look like the other compounds in the product or influencing the expected natural ratios between chemicals in the body.
So how do testing and certification organizations like NSF International test for these banned substances? Certified nutritional supplements are put through several complex and rigorous test procedures before they make it to store shelves. The main method for identifying and measuring potential banned substances in a supplement is liquid chromatography and gas chromatography coupled with mass spectrometry detection (LCMS and GCMS).
Mass spectrometry (MS) allows scientists both to identify the compounds in a supplement and to verify their quantity. The chromatography system allows separating the various components present in the sample. These components are passed to the mass spectrometry detector where the compounds are ionized and fragmented. The ions are then accelerated or deflected by electric and magnetic fields. Differences in the mass-to-charge ratio of the ions generate a total ion pattern that can be compared to reference compounds to confirm identity. To improve the sensitivity of the MS detector, specific masses that are most abundant for the compounds of interest are monitored, allowing for the detection of potential contaminants among the multitude of intentional ingredients. Using triple quadrupole or tandem mass spectrometers (LCMSMS or GCMSMS systems) allows for further selectivity by monitoring specific precursor ions as they transition to product ions.
A single compound may have several transitions. The uniqueness of the transitions for that chemical allows for the measurement of that compound in the presence of many other components. The frequency or intensity of the signal measured for the transition is proportional to the concentration present in the sample. A reference standard can be prepared at different concentrations to create a calibration curve and therefore the quantitative amount of a substance in a sample can be determined in comparison to the curve. But what if the analyst is looking for an unknown chemical?
Meet the LTQ Orbitrap. This type of liquid chromatography-mass spectrometry system offers an edge over the triple quadrupole LCMSMS systems for the detection of unknown chemicals due to its ability to measure the exact mass of the fragment ions. Typical LCMS systems have whole unit mass resolution, measuring the mass-to-charge of the ion to within ±1 Dalton (Da). The Orbitrap can discern very small mass differences to 0.004 Da with less than a two parts-per-million mass error or ±0.0008 Da. This is crucial because the difference between the mass of an acceptable compound and a banned substance sometimes comes down to a hundredth of the molecule’s mass.
The ability to identify modifications or structural changes of molecules is increasingly important as new designer drugs, steroids and steroid-like substances emerge before reference chemicals are available. Knowing the exact mass of the ions is key to determining which elements may be present in that ion and in what combination. In this way, the Orbitrap data can be used to elucidate chemical structure information for unknown chemical adulterants. The Orbitrap is most effective when unknown chemicals are present in samples at least at low part-per-million levels. This allows for adequate intensities of the ion peaks which results in the highest level of accuracy in the exact mass determinations and further confidence in the conclusions drawn from this data. Since banned compounds may be present in very minor concentrations, down to parts-per-billion levels, the instrument system needs to be well suited for the analysis goals. By combining Orbitrap technology for unknown determinations with triple-quadrupole (MSMS) systems for trace-level detection of known banned substances, the best of both worlds is achieved in ensuring that the products evaluated are free of these unwanted contaminants.
The use of new technologies and laboratory methodologies to test supplements ushers in a new era of contamination and banned substance detection. Instruments like the LTQ Orbitrap mass spectrometer allow programs like NSF Certified for Sport® not only to identify and measure compounds and banned substances in nutritional supplements, but also to identify new banned compounds as they emerge and to help spread awareness to the arena of testing and enforcement.