www.TheWercShop.com 310-703-9567 CannTrol Mold Kill Study Independent Report June 18, 2014 CONFIDENTIAL Table of Contents: 1 INTRODUCTION .......................................................................................................... 3 2 MATERIAL AND METHODS .......................................................................................... 4 3 RESULTS ..................................................................................................................... 5 3.1 3.2 3.3 4 LIVE SPORES CONTAINED IN INOCULATION SOLUTION ................................................................5 PLANT INOCLUATION TEST RESULTS ......................................................................................5 STRAIN PROFILE TEST RESULTS ............................................................................................5 CONCLUSION AND DISCUSSION .................................................................................. 6 1 Introduction Cannabis sativa L. is a plant that finds wide spread medicinal and recreational use. The main product from the plant is the dried flower of female plants. There are various cultivation, drying and curing conditions that may be encountered by cannabis cultivators in which mold contamination may become a significant problem. Standardized, rapid and effective means for product purity assurance are methodologies that are greatly needed in today’s rapidly evolving regulatory environment as new requirements for lower mold levels are being implemented by various US states and national governmental bodies around the world. The goal of this study is to investigate the effectiveness of the CannTrol Dry Curve to kill mold on cannabis. 2 Material and Methods Two days before harvest, two separate Cannabis plants of the same OG cultivar were placed inside of a single grow tent under one light. Other than isolation, these plants were treated exactly as any other plant in the normal course of cultivation efforts. Live aspergillus and botrytis mold spores were provided through resources of the client. These live spores were first enumerated and found to contain over 10ˆ6 CFU/g of solution. This verifies the spores arrived alive. Approximately 1.5g of solution was added to an isolated cola on the plant over such an area as to represent approximately 1g or less in dry weight when that cola would be completely dried. This estimate was made by the cultivator who had good experience in terms of what the wet and dry weight of this variety are in a typical cycle. A control sample, a CannTrol sample, and a blank sample were all pulled from the same plant. A control sample is one that was inoculated and then not placed inside of the CannTrol system, it was instead placed in a bag and allowed to stay at room temperature until being prepared for microbiological testing. The CannTrol sample was placed inside of the CannTrol system and cycled through a single cycle according to the manufacturer’s operating procedure. Blank meaning no inoculation solution was applied, this test being useful to determine there was no microbiological organism growth on the plant initially, before our purposefully doing so. 1g of dried flower material from each sample was ultimately collected and subjected to a standard mold screen performed in-house by The Werc Shop. Various CFU/g levels, from 10 CFU/g up through 100million CFU/g were prepared for each sample to explore the range of possible contamination. Standard cannabinoid and terpene profiles for the samples were also performed. 3 Results 3.1 Live Spores Present in Inoculation Solution An enumeration test was done on the initially received Aspergillus and Botrytis samples provided by the client and the results are shown below in Table 1. Fail means there were more than that level of spores present, pass means it was less than that level present. Table 1. CFU/g Tests of Spores Provided 3.2 Material 1,000 100,000 10,000,000 Aspergillus Spores Botrytis Spores FAIL FAIL FAIL FAIL PASS PASS Plant Inoculation Test Results Table 2 below shows the CFU/g test results of each of the inoculated samples studied, including those that were processed through the CannTrol system and those that were not (the controls). Flower that was present on the plants, but never inoculated directly and never placed through the CannTrol system was also tested to determine the presence of any other molds that may have been present but were not purposely placed on the sample material. Table 2. CFU/g Tests for Inoculated Samples Material Aspergillus - CannTrol Aspergillus – control Botrytis - CannTrol Botrytis – control Non-Inoculated Flower 3.3 10 100 1,000 10,000 100,000 PASS FAIL PASS FAIL FAIL PASS FAIL PASS FAIL FAIL PASS FAIL PASS PASS PASS PASS FAIL PASS PASS PASS PASS FAIL PASS PASS PASS Strain Profile Test Results Cannabinoid and terpene test results for the samples are shown below. A sample for both inoculations was taken in 2 parts, one for the material that went through the CannTrol system and the other for the control that was never placed in the CannTrol system. Table 3. Cannabinoid levels and sum of tested terpenes Material Aspergillus -CannTrol Aspergillus – control Botrytis – CannTrol Botrytis - control THC Max % THC % THCA % Terpenes 14.66 11.99 12.29 12.02 0.36 0.30 0.26 1.24 16.31 13.33 13.71 12.29 45.55mg/g 46.22mg/g 43.28mg/g 32.02mg/g 4 Conclusion and Discussion This study demonstrates the CannTrol mold kill methodology is effective at killing living mold present on cannabis at harvest. CannTrol appears effective at reducing levels of the two mold organisms studied down to below 10 CFU/g. The inoculation solutions were between 100,000 CFU/g and 10,000,000 CFU/g. We were informed it was around 10,000,000 and therefore could be reasonably assumed to be around 1,000,000 CFU/g. Applying 1 million CFU/g of inoculation solution in the fashion we used was effective at making molds grow on cannabis and these were detected on the plant material during the study. There were small amounts of mold present on the starting plant material before inoculation. CannTrol apparently killed whatever organism was present here as well. No significant change in cannabinoids or terpene profiles could be detected. The variations detected were most likely due to local variability in the plant and minor weight loss differences due to moisture content variability. Not enough samples were available to completely assess the local and total batch variability of the cannabinoid and terpene profile to be able to draw much more solid conclusions about this type of variability and the potential for how great it actually may be. Many more tests and samples would be required to be able to assess that much better. Overall the CannTrol system is incredibly easy to use and was demonstrated to be exceptionally effective at lowering the level of molds on cannabis plant material to below some of the most stringent regulatory standards being established in cannabis today.
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