June 16, 2015

LiftCannabis - A canadian leader for news on medical marijuana

Laurie Caron, Chemist, M.Sc. - Discoveries 
Since we obtained our Canadian licence to analyse medical marijuana, we connected to the businesses of this world. As with everything, news travel fast, and finding reliable references in this industry is not always an easy thing.

Some companies like LiftCannabis stand out more than others. Lift has a high presence in the social media and manage an interactive blog with a wide panel of contributors who are well-known in the industry. They write about various topics such as news on licensed producers, dispensaries, industries, patients, regulations, laws, etc. For this reason, I wanted to introduce you to this enterprise today on PhytoChemia's blog.

    Photo credit : http://liftcannabis.ca/

LiftCannabis is a canadian company from Vancouver, (BC) and I had the chance to discuss with David Brown, their communications director and I asked him some questions :  

Which are the reasons why Liftcannabis has started ?
"In 2013, when Health Canada began to formulate and implement its new regulations surrounding how Canadians access medical marijuana, it became clear to us that there was both misinformation and a dearth of accurate and useful information about the program (known as the MMPR), how it worked, who the producers were, etc.  So, we started Lift first and foremost as an information portal for Canadians looking to learn about medical marijuana and how to access it legally in Canada. Since then, Lift has grown into much more thanks to the participation from both patients and industry."

What kind of services do you offer ? Are they only for people who lives in BC or available to other provinces ? 
"Because Health Canada’s program is a federal one, our website offers a variety of information and services to all Canadians (although it’s presently only in English). We assist Canadians with learning about the benefits of medical marijuana and how to access the program.  We have listings of doctors from across Canada who are open to discussing medical marijuana as an option, and we also have detailed information about each Licensed Producer and their products. Furthermore, we have Canada’s largest database of legal medical marijuana strains and patient-generated reviews. In fact, we just recently logged our 1000th strain review. These are all real reviews left by real Canadian patients, and so they’re extremely helpful to all those who are searching for strains and trying to find the right one for their medical condition."

In May, Liftcannabis has organized a forum in Vancouver on medical cannabis. What was the main purpose of this event ? What kind of audience participated to this forum ?

"The purpose of our inaugural Lift Cannabis Forum in Vancouver this past May was to bring together different sectors of the medical marijuana industry to have an open and respectful conversation about what the future of cannabis regulation and standardization could look like in Canada. The audience was as diverse as our panel and included licensed producers, dispensary operators, home-growers, patients, doctors, lawyers, activists, and philanthropists.  It was really so rewarding to see the broad spectrum of Canadians being represented in that room, and hearing the opinions of everyone on how to work together to make this industry better for everyone."

In your blog, why did you include posts on research, quality control and science ?

"While it’s far from perfect yet, Canada’s federal medical marijuana program is the most advanced in the world in terms of regulation, standardization, and testing, and we feel sharing information about these aspects of the program is important. Patients can come to trust the sources and production methods of the medicine they consume, those perhaps sceptical of medical marijuana in general can get insight into how seriously regulated and strict the Canadian industry is, and those from other countries can take cues when establishing their own programs."

Do you plan any events in the next months?
  Photo credit : http://liftcannabis.ca/

"We have our second Lift Cannabis Forum scheduled for July 20 in Toronto.  It will once again feature a panel discussion, this time focusing largely on issues surrounding recent colleges of physicians guidelines and recommendations for doctors, the issue of marijuana ‘clinics’ as an avenue for access, the patient experience inside the MMPR, as well as looking at ways the MMPR could be refined to reflect changes to current pharmaceutical-grade standards.  More information and tickets can be found here: http://forum.liftcannabis.ca/toronto2015/ "

You can find Lift on Twitter, Facebook and Instagram also.

June 5, 2015

Tiguidou or How I Learned to Stop Worrying and Love the Analysis

About three years ago a friend of mine, who was doing her master on essential oils, was struggling with her analyses. This gave me the idea to automate part of the process by creating a small VBA program in Excel that would search in a database of rentention indexes for potential matches. She could then filter the ones that had the best MS match. Curious readers might wonder why the concurrent use of retention indexes is necessary along with MS: some information can be found in the previous post of my colleague Alexis St-Gelais on Retention indexes. He was aiming for at least three positive identification parameters: mass spectrum, polar retention index, and non-polar retention index. It did not works. There was too much variation between the RIs in literature due to method disrepancies and columns manufacturer (see here for an example).

Fast forward a couple of months later. We were offered to carry on the business of two local retired professors who were doing essential oil analyses for distillers (Dr Guy J. Collin and Dr François-Xavier Garneau). The method that they used for their analyses was the same that they were employing during their years as published researcher, and that we still use at Laboratoire PhytoChemia: dual column GC-FID analyses (as explained here). This method has some advantages (and some drawbacks, as for everything). One of them is that you have automatically two confirmations for the molecule identification (two retention indexes), as well as a validation on two columns for compounds concentrations, most of the time solving coelutions. You can easily get a third confirmation by using a MS when in doubt about identifications. The main challenge is that a very experimented analyst is necessary to be able to proceed efficiently. Given this, the idea of having something that would automate part of the job was still growing.

After a bit of programmation, Tiguidou* 1.0 was born. This Excel VBA software allowed us to simply paste the raw integration data in a spreadsheet and made an efficient first sorting for us. It would indeed automatically calculate the RIs on both columns, then, for every peak, search our database for a fitting non-polar RI and check if it was able to pair it with a polar RI. It would then order the results along a gradient of increasing discrepancy between theoretical pairs and observed values. The analyst would then choose from dropdown lists the best matches, or add one if needed. The program would then output a list ready to be copied in the report. This came along with a basic database management system, so we could add newly identified compounds as we found them from MS runs or literature.

*From an expression here in Quebec meaning "Everything's alright". Backronyms suggestions are welcome.

Tiguidou 1.0 Interface
Tiguidou 1.0 Database excerpt
We used this tool intensively for two years with a high success, since we cut the time lost in manually screening the databases by an order of magnitude. But, we felt that it still had some limitations. We decided that a new, more powerful version was needed. After almost 6 months of work, I am proud to announce that we are now working with Tiguidou 2.0. Tiguidou is no more confined to an Excel macro, it is now a web application, with a more dynamic database and a refined assignation algorithm. Version 2.0 can support more than two capillary columns, and tries to create the pairs in every possible way (not just non-polar->polar). It also checks if the percentages observed on both columns are similar to sort the results. The database is now built automatically from our previous reports, and now allows for a simple and full follow-up of which compound was seen where. In some way, it "learns" from each of our reports. A basic report, ready for final revision, is then automatically generated within our company template. We are also working hard on adding new options to the algorithm: to check if a compound was already observed in a given plant before, to develop statistics to automatically determine the expected range of each compound in an oil, to see if there is a possibility of latent chemotype, to correlate the presence of a compound to another, and many more. It may even eventually help us do the integration and peak picking!
Tiguidou 2.0 Interface
Tiguidou 2.0 Database excerpt
All this allows us to perform faster, more efficient, more complete and higher-quality analyses while always keeping a reasonable price and allowing more companies to have access to a good knowledge of their products. We plan to eventually make a public version that may allow producers and distributors to improve their quality control.

June 4, 2015

Question from the public: Phthalates contamination in essential oil

 Hubert Marceau, chemist, B.Sc.

I have recently been asked if it was possible that diethyl phthalate be naturally present in an essential oil, more precisely in a Cyperus scariosus sample where about 5% of DEP was detected. Here is our answer:

Bioaccumulation of DEP in plant.
There have been a nice review, a bit aged but very thorough, of the environnemental dispersion of phthalate in the environnement. Some studies made with Diethylhexyl phthatlate and Dibutyl phthalate have shown that plant grown in contaminated soil can bioaccumulate at a factor of 0.1% to 0.2%. Another study demonstrated the the range was between <0.001% and 1.0% with a typical value of 0.01%. I sadly can't find a clear value for DEP at the time. Also bear in mind that each plant has a different rate of accumulation for different product. I don't have the exact have value for Cyperus scariosus, but the genus Cyperus is used in bioremediation (removal of soil contaminant), especially for metal. I sadly also have no information on the bioaccumulation of phthalate ester. The phthalate accumulated would be distillated with the rest of the oil, it has a pretty high boiling point but some component with higher boiling (gaging on their retention index which is usually a function of the boiling point) do distill.

Cyperus scariosus essential oils yield is around 0.45% from a source, 0.5% to 1% from another and 0.077% to 0.80% from another. Let's use the best case scenario and go to about 0.5%. The specific gravity of the oil is around 1 so we'll leave it that way to simplify the calculation. That mean that to obtains 100 ml of the oil we need to distill 2 kg of plant. In this 100 ml of plant we have about 5% of DEP, which we will round to 5g. 5g in 2 kg is the equivalent of 2.5mg/g (please note that this level is very high, about 1000x more that what is usually found in litterature). If we assume that the bioaccumulation factor is  0.1 it mean that the concentration in the soil must be around 25mg/g. Reported value in polluted area in China and India are calculated ine µg/kg, about 1 000 000x time less than what is needed. If we use the more conservative yield of 0.077% we can divide the result by ten and it would still be 100 000x higher than what is usually report. We must also take account that a plant would probably saturate before that, it would not be able to have a concentration higher than an certain amount.

Now we have two situation, either:
a) we are in presence of a super phthalate bioaccumulator. That would very be suprising.
b) the phthalate was added after the distillation. More probable. By who? Who knows, it could be any one in the supply chain: the producer, the buyer, the wholesaler, the distributor, etc.

Cyperus Scariosus - Photo credit : Wikipedia

Natural presence of DEP in plant.
There have been report of phthalate in plant, most of the time it is due to contamination during the extraction process. I have learned quickly during my study to spot the double doublet at around 7.5 ppm in NMR and the ion at m/z=149 in MS, almost everyone in who studied natural product isolation once isolated pure phthalates (in the mg/kg range). They are often contamination from the lab solvent. There have been report of bacteria creating phthalates, but not DEP. Note that there are other "synthetic" compound that can be found in nature, for example blueberry contain parabens naturally.

Once more we are confronted to two situation, either:
a) we are in the presence of a good natural, and possibly industrial, source of DEP. That would be very surprising.
b) the phthalate was added after the distillation. See the Bioaccumulation of DEP part.

Concluding remark.
You will notice that I always give two potentials options, even if one is not very plausible. There is a reason for that: the Black Swan Hypothesis. In science you never have a 100% certainty, you can have 99.99999%, but it is impossible to have 100%, the same is true with proving that something is impossible. You would have to try every possible way to prove that something is impossible and this would require an infinite amount of time. Imagine the following, you are in a world where every swan is white. You conclusion is that for something to be a swan, it has to be white. Suddently you heard of an explorer who found a black swan. There are two option possible, your cognitive confirmation bias reject the idea, saying that the explorer might be wrong and that this may simply be a painted swan, or you change your definition. Now swan can be either white or black, this will stand true until somebody find a red swan. In which case you will have to chance your definition again.

Swans are black or white ? - Photo credit : Aliexpress

This allegory is true for essential oil too, we usually assume that an essential oil has a composition of X, based on analysis that were made. And this stand true, until somebody found an oil with a different composition. In the current market where more and more small and big producer are appearing, some who use local cultivar, it is my opinion that we will end up seeing more and more variation in the oil from a same species. This is specially true for oil distilled from plant stock harvested in the wild, or cultivated from wild specimen du to genetic diversity.

This allegory has not much to do with our current case, that seems to clearly indicate adulteration, but I wanted to introduce the concept of doubt and incertitude, which is a concept that I am very fan of, maybe not everybody will be of my opinion. I believe that have a full reflexion on something is often better than letting our emotion go (and getting our pitchfork). And on a final, more philosophical note, remember that your brain is wired to give you the impression that you are always right (confirmation bias), it actually take a conscious effort to see when we are wrong.


Staples, C.A., et al, The environmental fate of phthalate esters: A literature review, Chemospehere 35 (1997) 667-749
Zeng, F., et al, Phthalate esters (PAEs) Emerging organic contaminants in agricultural soils in peri-urban areas around Guangzhou, China, Environnemental Pollution 156 (2008) 425-434
Srivastava, A., et al, Occurrence of phthalic acid esters in Gomti River Sediment, India, Environnmenetal Monitoring Assessement 169 (2010) 397-406
Srivastava, R.K., et al, Chemical Constituents and Biological Activities of Promising Aromatic Plant Nagarmotha (Cyperus scariosus R. Br.): A review, Proceeding of the Indian National Science Academy 80 (2014) 525-536