Extraktlab 110 Cannabinoid Potency And Recovery 2d5r4e

931 Pine St, St. Croix Falls, WI, 54024

ExtraktLAB 110 Cannabinoid Potency and Recovery Summary The data presented in this paper demonstrates the ability of Extrakt-1 machines to produce high quality cannabinoid extracts. Samples taken over an extended period of time from several strains yielded similar results. With proper methodology, the Extrakt-1 machine can extract over 94% of available cannabinoids from plant material, with resultant extracts having an average of 77% cannabinoid potency.

CO2 Extraction A common method for the extraction of cannabinoids (essential oils) from plant matter uses pressurized CO2 as solvent. Cannabinoids and other material extracted from the plant material are suspended within pressurized CO2. As the pressure of CO2 is increased at a given temperature, the solubility of the cannabinoids increases dramatically but as the pressure or temperature is lowered, the solubilized wax and cannabinoid crashes out of solution. This leaves a cannabis extract with only small amounts of residual CO2 remaining, usually in the form of ice crystals. According to Prausnitz1, the process works well because the partial molar volume of the solubilized plant matter (terpenes, cannabis wax and cannabinoids) is a large negative number and is significantly larger than the molar volume of the pure solid.

State of the art CO2 Extraction Equipment Over the last several decades, industrial extraction machinery has been designed to manipulate temperature and pressure of CO2 for the efficient production of essential oils. The Extrakt-1 series, provided by extraktLAB, is a state-of-the-art CO2 extraction system developed to solve many practical and throughput problems that operators typically struggled to overcome. These problems included carry over, containment, reproducibility, throughput, and efficiency.

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931 Pine St, St. Croix Falls, WI, 54024

The Importance of Methodology Aside from the operational efficiency of the Extrakt-1, extraktLAB has developed a set of basic procedures that helps ensures the highest level of extraction efficiency. These procedures include proper drying, grinding plant material to a uniformed size, and decarboxylation of the material prior to extraction. The below results are taken from samples that followed that work path prior to processing in an Extrakt-1 machine.

Recovery of Cannabinoids from Plant Matter The following shows some typical results from a series of CO2 extractions conducted on a Extrakt-110plus on different lots of cannabis materials over several months. The cannabis lots were primarily sourced from trim but one lot used flower in addition to trim. Potency of the starting material does play a role in the resulting potency; this particular material assayed at 10-12% cannabinoids. Table 1. Demonstration of reproducible recovery of total cannabinoid collected from for five different lots of raw CO2 extracts over two months.

Test Date 4/8/2016 4/9/2016 3/11/2016 4/1/2016 5/6/2016 Average % Cannabinoid Stdev %

Sample Number 1 2 3 4 5

Batch Number 20160408 20160409 20160311 20160401 20160506

Total Cannabinoids (%) 77.3 81.4 75.3 77.3 76.5 77.6 2.3

a. Instrument Serial Number: 20160301 b. Total cannabinoids did not measure acids; potency is biased low. c. Third party testing conducted by Pure Analytics d. All extracts were sampled from unprocessed, non-dewaxed CO2 extracts.

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931 Pine St, St. Croix Falls, WI, 54024

First, the data clearly show that the potency of the raw and unprocessed recovered oil can be quite high even for CO2 extractions. This can be attributed to the fact that the ExtraktLAB methods used in this extraction did not over-extract the plant matter, resulting in a diluted extract,. Cannabinoids measured included THC, CBD, CBN, THCV, etc. These particular results also show that the extraction performance is consistent over time despite the fact that the lots of input material are changing. The following figure is a graphical representation of the data presented in Table 1. 100

% Cannabinoid Potency

90 80 70

77.3

81.4

75.3

77.3

76.5

3

4

5

60 50 40 30 20 10 0

1

2

Sample Number Figure 1. Percent recovered cannabinoid from raw CO2 extracts for five distinct lots of extracted materials over the period of 2 months.

Measurement and Calculation of Recovery There are two common ways to measure the recovery of an extraction. These methods include determining the weight recovery of oil or determining the weight loss from the extracted plant matter. Both methods are useful but, there are specific advantages and disadvantages to each . The first method involves weighing the plant matter (with a calibrated balance) before it is placed into the extractor, and weighing the oil that is recovered from the collector. The recovery is calculated with the following formula: %𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅(𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵) = 100 ∗ 3

𝑊𝑊𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑜𝑜𝑜𝑜𝑜𝑜

𝑊𝑊𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖 𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚

931 Pine St, St. Croix Falls, WI, 54024

This method, although very simple, is the most imprecise of all the methods. Sources of variation include: • • • • • • • •

the water content of the plant matter before it is extracted the impossibility of quantitative transfer of a waxy extract from one vessel to another the difficulty of thorough and adequate cleaning of the collection vessels to ensure full recovery of the extract off of the walls, tubing and other internal parts of the machine the presence of process aids used for cleaning (such as ethanol).. the presence of dry ice the portion of extracted plant matter that is left in the recycled co2 or vented to atmosphere improper calibration or balance range for the measurement. different strains with differing oil content.

One critical observation to note here is that the potency of the extract is irrelevant to this number. For example, in over extraction conditions, a higher weight recovery of the plant can occur at the cost of potency. It is up to the process engineer to ensure that relevant recovery parameters are reported and utilized by the business to manage the process. These sources of variation can lead the operator to false conclusions about recovery if they are not properly controlled. For example, if the water content of the plant matter is unknown and changing, the weight recovery will vary in proportion. In general, this method should be avoided if process analytics are available. However, if a balance is all that is available for recovery calculations, it would be preferable for the plant matter to be weighed before and after extraction. %𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅(𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵𝐵) = 100 ∗ �1 −

𝑊𝑊𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎 � 𝑊𝑊𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖 𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚

In this case, items 2-6 are eliminated as sources of variability and a more consistent bulk recovery number is measured.

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931 Pine St, St. Croix Falls, WI, 54024

A superior and more consistent way to measure recovery is by measuring the cannabinoid content of the plant matter before and after extraction. This approach has many advantages but is also highly dependent on the skill of the analyst. Advantages include: • • • •

measurement of cannabinoid content of plant matter before extraction allows one to measure the recovery of individual cannabinoids methods can be specifically tuned to the cannabinoid content thereby improving throughput consistent and reliable measurements measurement of total cannabinoid content.

Pitfalls include: • • •

analytical proficiency requirements lack of recovery from improper sample preparation the use of methods that are not validated.

This technique involves sampling the plant matter and properly measuring the cannabinoid profile by measuring the peak area for each cannabinoid under the curve in a chromatogram. The material is then sampled after extraction and the same method is repeated. The cannabinoid content can be summed but should be reported as a potency or total cannabinoid recovery. The recovery is calculated according to the following equation. %𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅(𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻) = 100 ∗ �1 −

𝑃𝑃𝑃𝑃𝑃𝑃𝑃𝑃 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎 � 𝑃𝑃𝑃𝑃𝑃𝑃𝑃𝑃 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖𝑖 𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚

An alternative to this approach is to calibrate the HPLC and report either the weight recovery of each cannabinoid or the total potency.

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931 Pine St, St. Croix Falls, WI, 54024

The following table shows the percent recovery as measured by HPLC for a series of lots of material. Table 1. Recovery percent of total cannabinoid collected from for five different lots of plant material. Sample Number 1 2 3 4 5 Average % Recovery Stdev %

Total Cannabinoid Recovery (%) 95.1 96.7 92.9 97.1 90.2 94.4 2.9

a. Instrument Serial Number: 20160301 b. Total cannabinoids did not measure acids; potency is biased low. c. Third party testing conducted by Pure Analytics

Run Times for Collected Data All extractions analyzed and compared within were processed in 1 hour run times. Extrakt-1 machines incorporate a chart recorder within their software system. The below graphics illustrate a typical work day producing the analyzed samples.

Typical raw product 6