HerbalEGram: Volume 10, Number 2, February 2013

The controversy continues over the origin of DMAA, the chemical otherwise known as 1,3-dimethylamylamine or methylhexanamine (MHA). Proponents of the ingredient — namely, manufacturers and marketers of supplements containing DMAA — claim that it is a naturally occurring constituent of the oil, leaves, and/or stems of certain types of geranium (Pelargonium graveolens), and is therefore a legal dietary ingredient. These assertions have been largely debunked, however, as discussed in an article in HerbalGram 95, which detailed the results of two independent scientific papers from university laboratories that failed to detect any presence of the chemical in authenticated geranium plant material.^1-3 Since then, two papers funded by a leading DMAA-containing supplement producer* and published in Analytical Chemistry Insights — an open-access, online-only journal — have claimed to detect miniscule quantities of the chemical in geranium plants from various regions in China.^4,5 Analytical chemistry experts, however, question the authenticity of the plant samples used in these studies, among additional factors that potentially affect the significance of the findings.


International Attention

The safety of the amphetamine-like substance was brought into question after the deaths of two US military service members in December 2011. Autopsies found in their systems traces of DMAA, which is found in some popular pre-workout supplements. Shortly after, the Department of Defense urged the removal of all DMAA-containing supplements from military commissaries and exchanges as a precaution.⁶ A year earlier, a New Zealand medical journal reported a case of a brain hemorrhage in a 21-year-old male who took a party drug containing DMAA and caffeine, in addition to consuming alcohol.⁷ More recently, in April 2012, 30-year-old Claire Squires collapsed and died in the final mile of the London Marathon. According to a January 2013 coroner’s report, the cause of death was “cardiac failure caused by extreme exertion, complicated by DMAA toxicity.”⁸

Although DMAA has been on the World Anti-Doping Agency’s (WADA) list of banned substances since 2010, various countries’ supplement-regulating bodies have been slower to respond to the situation.⁹ On April 27, 2012, the US Food and Drug Administration (FDA) sent warning letters to 10 dietary supplement manufacturers that produce supplements with the controversial ingredient, warning that synthetically produced DMAA is not a “dietary ingredient’” and is therefore not permitted to be used as a component of dietary supplements.¹⁰

The 10 warning letters — one of which was sent more recently, on August 28, 2012 — cited the companies for failing to file a New Dietary Ingredient (NDI) notification for DMAA, and for not providing FDA adequate safety data for the chemical, as required under Section 8 of the Dietary Supplements Health and Education Act of 1994 (DSHEA). Such omissions render the products adulterated, and the introduction of adulterated products into interstate commerce is a prohibited act under the governing sections of the US Federal Food, Drug, and Cosmetic Act.¹⁰

In the warning letters, the FDA noted that “DMAA is known to narrow the blood vessels and arteries, which can elevate blood pressure and may lead to cardiovascular events ranging from shortness of breath and tightening in the chest to heart attack.” According to the letters, the FDA had received 42 adverse event reports related to DMAA-containing products. The FDA’s website states, “While the complaints do not establish that DMAA was the cause of the incidents, some of the reports have included cardiac disorders, nervous system disorders, psychiatric disorders, and death.”¹⁰

The idea that DMAA was a naturally occurring constituent of geranium stems from a single paper by Ping et al. published in 1996 in the now-defunct Journal of Guizhou Institute of Technology.^12,13 The paper was intended to be a simple catalog of the constituents of rose geranium oil, and the occurrence of DMAA and several other amines in the oil was unremarked upon by the author. The amines are listed in a long table, but are otherwise not mentioned in the article. Numerous typographical and translational errors (the compound widely reported by advocates to be DMAA based on the English translation of the Mandarin characters had an incorrect molecular formula) also marred the report, and the findings in the paper are contested in the scientific community.^14,15 In Health Canada’s DMAA classification document, for example, it stated that “Given the errors in the Ping et al. (1996) paper and that the fact that these authors did not fully isolate and characterize DMAA, together with the contradictory information from numerous other peer-reviewed scientific studies which show no natural occurrence … there is no credible scientific evidence that DMAA is captured as an isolate of a plant … and therefore cannot be classified as [a natural health product].”¹⁵


Laboratory Findings

More than a year after the US military deaths, the controversy continues in the scientific literature. In the summer of 2012, papers by ElSohly et al. in the Journal of Analytical Toxicology and by Zhang et al. in Drug Testing and Analysis found no detectable traces of DMAA in authenticated geranium samples with extremely sensitive limits of detection of 10 parts per billion.^2,3

Lead author Mahmoud A. ElSohly, PhD, a research professor in the Research Institute of Pharmaceutical Sciences at the University of Mississippi, calculated the amount of geranium plant material it would take to make one capsule of a DMAA-containing supplement using estimates established by the now-infamous Ping et al. paper.¹³

“If MHA were to be present in the oil at 0.6% (i.e., 6mg MHA/mL [geranium] oil) as reported [in the paper by Ping et al.], … it would require approximately a kilogram of leaves to be harvested to prepare 6mg of MHA,” he wrote in the paper.² “Elementary mathematics establishes that the MHA in dietary supplements must be synthetic.”

In the Zhang et al. paper published in July 2012, researchers from the University of Texas at Arlington used a unique chemical property of DMAA to determine its natural or synthetic origin.³ Due to certain properties of its atoms, DMAA exists in four potential spatial arrangements — the study of which is referred to as stereochemistry. Zhang et al. analyzed and compared the stereoisomeric ratios of synthetic DMAA standards and from DMAA used in commercially available supplements. They found that the composition of DMAA in supplements was identical to that of synthetically produced DMAA, concluding that the DMAA found in the supplements they analyzed must be synthetic, and could not be from any plant or other natural origin.³


Industry-Funded Studies Lack Adequate Authentication

More recently, Li et al. and Fleming et al. published papers purporting to document the presence of DMAA in nanogram per gram (ng/g) quantities in certain geranium plants from various regions in China (A nanogram is 0.000000001 grams or one billionth of a gram). In addition, both papers analyzed the diastereomeric ratios of 1,3-DMAA, in an attempt to elucidate the origin of the chemical.^4,5 Interestingly, the papers also looked for the presence of 1,4-DMAA, an isomer (i.e., a compound that has the same chemical formula but a different structure) of 1,3-DMAA that is not known to be a component of dietary supplements.

The Li et al. and Fleming et al. papers, both published in Analytical Chemistry Insights, received funding from USP Labs, LLC, of Dallas, Texas, which manufactures the DMAA-containing supplements Jack3d^® and OxyElite Pro^® that are now the subject of numerous class-action lawsuits.¹⁷ The venue in which the reports were published, in addition to the funding source, have raised questions in the minds of some independent experts and analytical chemists.

“All the USP Labs-funded papers have been published in open-access journals, meaning [the company] paid for publication,” stated Mark Roman, PhD, an analytical chemist who is president and technical director of Tampa Bay Analytical Research (email, January 9, 2012). “Open-access journals have notoriously poor peer-review processes. The fact that only USP Labs-funded studies published in open-access journals have shown the presence of DMAA in geranium oil does raise my suspicions.”

In fact, according to the Analytical Chemistry Insights website, North American authors submitting articles for publication must pay the journal a fee of $1,848.^18,19

Li et al. paper

Li et al., of Intertek-AAC Labs in Champaign, Illinois, used a specialized type of liquid chromatography — known as electrospray ionization/tandem mass spectrometry (LC-ESI/MS/MS) — to determine the quantities of 1,3- and 1,4-DMAA in synthetic chemical reference standards, geranium plants, and geranium oils. The standards were purchased from Sigma-Aldrich, a supplier of biochemical and organic chemical products and services in St. Louis, Missouri.

According to the paper, geranium plants from three areas in China were provided by Yi Jin of Yunnan University and were authenticated by Professor Xu Youkai, PhD, of the Xishuangbanna Tropical Botanical Garden-Chinese Academy of Sciences. Geranium oil samples were acquired from Jiangxi Ji’an Hengcheng Flavor Oil Factory and stored at room temperature.⁴ No additional information on authentication methods was provided, and this lack of documentation has raised concerns among various analytical chemistry experts as to the authenticity and validity of the samples.

After analyzing the diastereomeric ratios, the authors found that “1,3-DMAA has two isomer peaks which are present in equal amounts and which are identical in all tested samples, including the synthetic reference,” they wrote. “Most likely only one chiral configuration would be present in plants (often referred to as the natural form). The results in the current study show that 1,3-DMAA in geranium plants and geranium oils appears to be an exception to this notion.”⁴

Detection methods revealed the presence of 1,3-DMAA in all tested plant and oil samples, with quantities ranging from 13.6 ng/g to 13271 ng/g (Table 5 in the Li et al. paper). 1,4-DMAA was also detected in some plant and oil samples, but not all. “Therefore, to our knowledge, the present study is the first to show conclusively that DMAA is naturally occurring in geranium plants,” the authors wrote.⁴

Fleming et al. paper

The most recent paper on DMAA was published on November 9, 2012, also in the journal Analytical Chemistry Insights. Fleming et al., of the department of chemistry at the University of Memphis, used liquid chromatography-tandem mass spectrometry (LC-MS-MS) to determine the presence and quantity of 1,3- and 1,4-DMAA in geranium plants from three regions of China — Changzhou, Guiyang, and Kunming — collected in winter and summer months. Diastereomeric ratios of 1,3-DMAA in the plants were measured as well and compared with synthetic standards.⁵

Synthetic chemical reference standards were purchased from 2A Pharmachem USA in Lisle, Illinois and Sigma-Aldrich, and plant samples were collected and authenticated by the same individual used in the Li et al. paper. No further details of authentication methods or chain of custody were provided.⁵

As noted above, the lack of information on plant sample authentication in both of the recent papers is concerning to some experts. “The Fleming et al. paper shows no independent authentication of the materials, nor chain-of-custody,” Dr. Roman stated. “In the Li paper, the plant materials were all authenticated in China with no additional authentication performed by their laboratory. This raises concerns about the possibility of adulteration.”

Dr. ElSohly echoed concern about the samples’ authenticity and purity. “This is what they say they found,” he said. “Whether what they found is actually there, or whether the materials that they received were contaminated, I don’t know. I can say what they are presenting seems like the work itself is okay. Of course, the results are only as good as the samples you are analyzing.”

Fleming et al. found that “neither the winter nor summer harvest samples of Kunming and Guiyang samples contained 1,3-DMAA or 1,4-DMAA species above the MDLs [method detection limits] of the analysis. However, Changzhou samples resulted in similar concentrations of 1,3-DMAA and 1,4-DMAA in the [summer] and [winter] 2011 samples.”⁵

Chromatographic analysis showed that “the results of the geranium plant diastereomer ratios are similar to the ratios of the synthetic standards presented here, as well as the standards and supplements analyzed by Zhang et al,” they wrote. Curiously, the authors concluded that “this indicates that supplements containing both 1,3-DMAA diasteromer pairs could be naturally produced and extracted from geranium plants.”⁵ This is, essentially, the opposite of the conclusion made by Zhang et al.

This finding has raised questions in the minds of scientists familiar with DMAA. Daniel W. Armstrong, PhD — the Robert A. Welch Chair in Chemistry at the University of Texas at Arlington, a recognized expert in the chiral chemistry of natural products, and co-author of the Zhang et al. paper — explained his concerns.

“It’s just really unlikely that you’re going to have exactly the same stereoisomeric ratio [in supplements and plant materials],” he said (oral communication, January 8, 2013). “Why are they all exactly the same as the synthetic standards? Seasons and nutrients, and the weather [they were] grown in, and the soil [they were] grown in, change various molecules and ratios of molecules in plants including some stereoisomers, but for some reason, these always seem to be the same as the synthetic. That seems to me to be a little odd.”

Dr. ElSohly shared Dr. Armstrong’s concerns. “In this particular case, we have a compound that has two asymmetric centers, which result in four stereoisomers. For [all isomers] to have exactly the same ratio in a natural product, that is extremely unusual.”

Dr. Roman opined that the publication of these recent papers might be for non-scientific reasons. “The industry, particularly USP Labs, is using DSHEA as an excuse to [try to] justify keeping DMAA on the market,” he stated. “By ‘proving’ that DMAA is present in geranium oil, even at the ppb [parts per billion] level, the argument is that an NDI does not need to be filed. [These are the levels] you might expect to find [in] things like pesticides, heavy metals, polyaromatic hydrocarbons, etc., and below what would be considered a pharmacologically active level. Nobody would argue that consuming multi-mg quantities of any of these contaminants is safe, even though they were in the food supply at ppb levels for a long time. This is certainly not in the spirit of DSHEA.”

The papers by Li et al. and Fleming et al. are the first since the controversial 1996 Ping et al. paper purporting to have found quantities of DMAA in geranium and could have significant ramifications for the DMAA debate.¹⁵

“When you have an exceptional find in science, something that’s extraordinary, then you need extraordinary or exceptional evidence and proof,” said Dr. Armstrong.


The Dim Future of DMAA

The experts contacted for this article consented that the heyday of DMAA is over. Although Jack3d is still available online from USP Labs, GNC.com, and VitaminShoppe.com, USP Labs reformulated a DMAA-free version of the product that it calls Jack3d micro™.²⁰ Many other supplement manufacturers are following suit and DMAA-containing products are becoming increasingly difficult to find.²¹

According to an article from Nutraingredients-USA.com, Joe Fortunato, CEO of General Nutrition Centers, Inc. (GNC), was quoted as saying, “The pre-workout category is still strong but it is not being driven by DMAA products. We have made a very concerted effort to move away from DMAA products in our stores.” He continued, “Rather than keeping the controversy going on DMAA it seemed like an easier thing to do as replacement products have been very effective and did not have the so-called tarnished effect of DMAA.”²²

The food-, health-, or drug-governing bodies of a growing number of countries have issued warnings on DMAA. In March 2012, Australia added DMAA to its Schedule 9 list of prohibited substances, which also included cannabis (Cannabis sativa), heroin (derived from the opium poppy, Papaver somniferum), and LSD. Consumer warnings reportedly have been issued in many other countries, including Canada, the United Kingdom, Australia, most Scandinavian countries, Ireland, Germany, Spain, and other European nations.^23,24

The two new USP Labs-funded studies have ensured that the DMAA debate will continue to play out in the scientific literature. “It’s added some confusion there, putting some doubt on whether it’s natural or not,” said Dr. ElSohly. “Of course, in my own mind I have no doubt that the DMAA that’s in the dietary supplements is not natural, but some might think otherwise and they are free to think the way they want to think. I’m not sure we’ve seen the last of the debate yet.”


* Both Li et al. and Fleming et al. acknowledge that their papers were financially supported by USP Labs, the company that sells Jack3d^®, Jack3d Micro^®, and OxyElite Pro^®. In the “Competing Interests” section of the Li et al. paper, the authors “disclose that funding for analytical research and manuscript preparation was provided by USPlabs, LLC” and that “ZCL [a co-author] served as an expert witness in 2011 for the case: DeRosier v. USPlabs.” Additionally, “The sponsor initiated a request to this laboratory to investigate the presence of DMAA in geranium plant and geranium oil. … Data analysis and the manuscript preparation were performed by the authors of the manuscript, while the sponsor provided grammatical review and assistance.”


—Tyler Smith


References

1. Smith T. New research suggests synthetic origin of DMAA in supplements. HerbalGram. 2012; (95):46-49. Available here. Accessed January 7, 2013.

2. ElSohly MA, Gul W, ElSohly KM, et al. Pelargonium oil and methyl hexaneamine (MHA): analytical approaches supporting the absence of MHA in authenticated Pelargonium graveolens plant material and oil. Journal of Analytical Toxicology. 2012:00:1-15. Available here. Accessed January 7, 2013.

3. Zhang Y, Woods RM, Breitbach ZS, and Armstrong DW. 1,3-dimethylamylamine (DMAA) in supplements and geranium products: natural or synthetic? Drug Testing Analysis. 2012:4(12):986-990. Available here. Accessed January 7, 2013.

4. Li JS, Chen M, Li ZC. Identification and quantification of dimethylamylamine in geranium by liquid chromatography tandem mass spectrometry. Analytical Chemistry Insights. 2012:7:47-58. Available here. Accessed January 3, 2013.

5. Fleming HL, Ranaivo PL, Simone PS. Analysis and confirmation of 1,3-DMAA and 1,4-DMAA in geranium plants using high performance liquid chromatography with tandem mass spectrometry at ng/g concentrations. Analytical Chemistry Insights. 2012:7:59-78. Available here. Accessed January 3, 2013.

6. Daniells S. Dept of defense supports removal of DMAA products from military facility shelves. Nutraingredients-USA website. Available here. Accessed January 4, 2013.

7. Starling S. DMAA “party pills” blamed for cerebral haemorrhage in NZ man. Nutraingredients website. Available here. Accessed January 4, 2012.

8. Claire Squires inquest: DMAA was a factor in marathon runner’s death. BBC News website. Available here. Accessed January 31, 2013.

9. A DMAA scandal timeline. NewHope360 website. Available here. Accessed January 4, 2013.

10. FDA challenges marketing of DMAA products for lack of safety evidence [press release]. April 27, 2012. Silver Spring, MD: US Food and Drug Administration. Available here. Accessed January 4, 2013.

11. Dietary supplement health and education act of 1994. US Food and Drug Administration website. Available here. Accessed January 4, 2013.

12. Shipley A. Chemists new product contains hidden substance. The Washington Post. May 7, 2006. Available here. Accessed January 30, 2013.

13. Ping Z, Jun Q, Qing L. A study on the chemical constituents of geranium oil. Journal of Guizhou Institute of Technology. 1996. 25(1):82-85.

14. Link C. Why the new USPlabs-funded DMAA study came too late. New Hope 360 website. Available here. Accessed January 14, 2013.

15. Classification of 1,3-Dimethlyamylamine (DMAA). Health Canada Health Products and Food Branch website. Available here. Accessed January 15, 2013.

16. Stereochemistry: chiral molecules. Wiley website. Available here. Accessed January 8, 2013.

17. Watson E. Allmax Nutrition, BPI Sports, and DynaPep Corp targeted in latest wave of class action suits over DMAA. Nutraingredients-USA website. Available here. Accessed January 7, 2013.

18. Article processing fees. Libertas Academia website. Available here. Accessed January 14, 2013.

19. Beall J. Beall’s List of Predatory, Open-Access Publishers: 2012 Edition. Available here. Accessed January 29, 2013.

20. Jack3d Micro™. USP Labs Direct website. Available here. Accessed January 8, 2013.

21. Watson E. Coming soon: Jack3d Micro (minus the DMAA). Nutraingredients-USA website. Available here. Accessed January 4, 2013.

22. Watson E. GNC backpedals on DMAA and prepares launch of ‘banned-substance-free’ MARKED range. Nutraingredients-USA website. Available here. Accessed January 7, 2013.

23. Starling S. Australian medicines agency adds DMAA to poisons list. Nutraingredients-USA website. Available here. Accessed January 4, 2013.