Monday, October 27, 2014

The Dendrobium Craze, Or What Happens if You Ignore the Obvious Difference Between "A" and "B" - Latest Study: At Best Trace Amounts of "Meth Analog" in DS Craze

DS Craze was not the only dendobrium containing supplement that disappeared from the market, when "the craze" about meth in dietary supplements broke lose.
You will certainly remember the craze about Craze and other dendrobium based pre-workout supplements. In an article published in Drug Testing and Analysis on October 14, 2013, scientists from Harvard Medical School and NSF International, a respected third-party testing organization, reported the presence of the "methamphetamine analog" N,α-diethyl-phenylethylamine (N,α-DEPEA) in three samples of Driven Sports' flagship pre-workout product "Craze" they purchased from various retailers.

Using what they considered established reference standards and sensitive analytical methods, Cohen et al. found what they thought was N,α-DEPEA at concentrations ranging from 21 to 35 mg per serving in the supplements (Cohen. 2013)
Citrulline is another pre-workout ingredient - a 100% safe one ;-)

Citrulline prevents muscle catablism more than leucine

Arginine & citrulline for blood lipid control

EAA, BCAA, or citrulline for anti-catabolism?

Glutamine not citrulline to heal the gut?

Citrulline to ignite fatty acid oxidataion?

High & low dose arginine ineffec- tive NO boosters
The results from this the study Cohen et al. conducted in 2013 were later officially confirmed by researchers from the Korean Forensic Service who found 0.40 and 0.44% N,α-DEPEA, respectively — the equivalent of roughly 23 mg per serving in their samples of Craze (Lee. 2014).

Now, N,α-DEPEA is only one out of several phenylethylamines aka "PEAs", of which even Cohen et al., whose paper started the whole craze say that the spectrum of PEAs "range[s] from benign compounds found in chocolate to synthetically produced illicit drugs" (Cohen. 2014). In that, minimal structural differences determine, whether we are dealing with a harmless ingredient of chocolate or a methamphetamine analog like N,α-DEPEA with the difference in the chemical structure of the agents being small, in some cases marginal (Lee. 2014).
Figure 1: Different phenylethylamines (PEAs) with different degrees of structural resemblance
and different physiological effects (Wahlstom. 2014)
In this context, it's important to know that N,α-DEPEA is not the only isomer of diethyl phenethylamine. Next to N,α-diethyl phenethylamine (the N,α isomer), there are also N,β-diethyl-phenethylamine (the N,β isomer), and N,N-diethyl-phenethylamine (the N,N isomer). Three agents of which Dallas Wait writes in a recent issue of the AHPA Report that specifically the...
"[...] N,α and N,β isomers have very similar chromatographic characteristics and are difficult to resolve unless the test method being used is optimized to properly separate both isomers" (Wait. 2014)
This similarity makes the two DEPEA isomers very difficult to distinguish. More importantly, the existence of the N,β isomer implies that anyone who analyzes dietary supplements for N,α-diethyl phenethylamine runs the risk of generating false positive  - yet unfortunately, this is exactly what Cohen et al. (2013) and Lee et al. (2013) and before them Mahmoud A. ElSohly & Waseem Gul (2013) obviously did.
My previous Craze article is now obviously obsolete | read more.
"As such, the alleged presence of the N,α isomer may actually be the N,β isomer, or a combination of the two.

Unequivocal identifiation of the N,β and N,α isomers is particularly critical since the N,β isomer does not share the same structural similarities that N,α shares with methamphetamine (also alkylated in the α position, not the β position)." (Wait. 2014)
Needless to say that the existence of said structural difference does also imply that the agent won't fit into the corresponding receptor site or interact with the same enzymes N,α-DEPEA and "real" methamphetamies do. Accordingly, the biological activity associated with that compound are certainly different from those of N,α-diethyl phenethylamine.
What is phenethylamine? Phenethylamine (PEA) is a natural monoamine alkaloid that belongs to a class of chemicals with many compounds of known psychoactive and stimulant effects (Glen. 2005). It functions as a neuromodulator or neurotransmitter (Sabelli. 1976) with similar actions as amphetamine: Norepinephrine + dopamine release (Parker. 1988; Paterson. 1993). PEA and its substituted forms NDP and ETH is rapidly metabolized when taken orally (Shulgin. 1995-2009).
Obviously I wouldn't be telling you all this if it wasn't related to the (un?)surprising results of a recent study. Results a group of researchers from Q&Q Labs AB in Gothenburg, Sweden, published in issue #6 of the peer-reviewed journal Analytical Methods that is published by the Royal Society of Chemstry (Wahlstom. 2014).

In said paper, the authors, Wahlstrum, Styles and Hägglund present the results of an analysis of two samples of Driven Sports' pre-workout product Craze ("Candy Grape", batch # 1305323 and "Berry Lemonade" batch # 1303298) that was conducted with a new analytical method that can "reliably" identify and quantify the individual amounts of N,α-diethyl phenethylamine (N,α isomer), N,β-diethyl-phenethylamine (N,β isomer), and N,N-diethyl-phenethylamine (N,N isomer) in a sample.
Table 1: Amounts and relative standard deviations (RSD) for the two samples of
DS Craze with lot no. 1303298 and 1305323 (Wahlstrom. 2014)
Usually I would plot the results, but in view of the fact that the amount of N,α-diethyl phenethylamine that was previously reported to be in the milligram range is de facto in the nanogramm range (1/1000 mg = 1 µg = 1000 ng), I decided to stick to the tabular overview (Table 1) from the original publication by Wahlstrom et al. (2014).
Figure 2: Mean amount of N,α-diethyl phenethylamine (in ng) in one serving of Craze according to Wahlberg (2014) and Lee (2014) - mind the logarithmic scaling of the graph!
So what does all that mean? You don't have to be a mathematics professor to see that even with the calculated standard deviations of 6.6% and 8.9% we are miles away (103303% to be precise) from the previously reported 23mg of N,α-diethyl phenethylamine per 5.8g serving of Craze (Lee. 2014).

This does not necessarily mean that all the craze was for nothing, though. As far as I know it is likely that N,β-diethyl-phenylethylamine is harmless. Studies which would investigate its biological effects on rodents, let alone animals, on the other hand, are as of yet not available; and if we are honest this and not the presence of "structural analogs" of whatever should be what we as supplement users are interested in, right?

Assuming that Driven Sports was willing to pay not just for the study at hand which was "made possible", as the authors of the study say, "by support from Driven Sports Inc" (Wahlstrom. 2014), but also for an analysis of what exactly the major DEPEA compound in the product does, this may finally solve the whole conundrum. A safety analysis for the complete supplement is - at lest for short-term use - available, anyway (Kedia. 2014) | Comment on Facebook!
  • Cohen, Pieter A., John C. Travis, and Bastiaan J. Venhuis. "A methamphetamine analog (N, α‐diethyl‐phenylethylamine) identified in a mainstream dietary supplement." Drug testing and analysis (2013). 
  • ElSohly, Mahmoud A., and Waseem Gul. "LC–MS-MS Analysis of Dietary Supplements for N-ethyl-α-ethyl-phenethylamine (ETH), N, N-diethylphenethylamine and Phenethylamine." Journal of analytical toxicology (2013): bkt097. 
  • Kedia, A. William, et al. "Effects of a Pre-workout Supplement on Lean Mass, Muscular Performance, Subjective Workout Experience and Biomarkers of Safety." International journal of medical sciences 11.2 (2014): 116.
  • Lee, Jaesin, et al. "Identification and quantitation of N, α-diethylphenethylamine in preworkout supplements sold via the internet." Forensic Toxicology 32.1 (2014): 148-153.
  • Wait, D. "The importance of reliable product chemistry data: A case study." In AHPA Report | The Official Publication of the American Herbal Products Association 29.10 (2014). 
  • Wahlstrom, R., C. Styles, and G. Hägglund. "Reliable identification and quantification of three diethylphenethylamines in a Dendrobium-based dietary supplement." Analytical Methods 6.19 (2014): 7891-7897.