Source: CONNECTICUT AGRICULTURAL EXPERIMENT STATION submitted to NRP
INDUSTRIAL HEMP PRODUCTION, PROCESSING, AND MARKETING IN THE U.S.
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1023295
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
S-OLD 1084
Project Start Date
Jun 4, 2020
Project End Date
Sep 30, 2023
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
CONNECTICUT AGRICULTURAL EXPERIMENT STATION
PO BOX 1106
NEW HAVEN,CT 06504
Performing Department
Analytical Chemistry
Non Technical Summary
The 2018 farm bill removed hemp, defined as Cannabis sativa, with less than 0.3% delta-9 tetrahydrocannabinol (THC), from the schedule I controlled substances list. Cultivation of hemp began in Connecticut (CT) in 2019. Hemp growers in CT have primarily focused on the production of cannabidiol (CBD) which is in high demand for its potential role in easing symptoms of many common health issues, including anxiety, depression, acne and heart disease. In other areas of the country hemp may be grown for seed or oil. There is the possibility that pesticides may intentionally or unintentionally be applied during the production of hemp for CBD, seed or oil. The goal of this work is to determine to what extent pesticide residues end up in products intended to be ingested by the consumer.The Connecticut Agricultural Experiment Station (CAES) is an International Organization for Standardization (ISO) 17025 accredited laboratory for pesticide residue analysis in food. It has worked with the CT Department of Consumer Protection (CT DCP) since the 1963 and is the de facto laboratory to provide regulatory enforcement of pesticide residues in food sold within the state. Since 2019 the CAES has worked collaboratively with the CT Department of Agriculture (CT DoAg) to test hemp grown in the state for THC content prior to harvest. The CAES methodology used for food testing will be applied to the testing of Cannabis sativa bud, leaf and seed. The method will be validated to first ensure that pesticides of interest can be appropriately tested in the hemp matrix. Work will then examine how effectively pesticide residues are co-extracted during the purification of the CBD, and if they ultimately end up products available to the consumer.
Animal Health Component
0%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71117302000100%
Goals / Objectives
Agronomic practices -Determine effects on grain, fiber, or dual-purpose productivity as functions of * Cultivars - including suitability to growing conditions/regions. This entails evaluating and developing adapted, improved, monoecious cultivars for grain, fiber, dual purpose (grain + fiber), and essential oil production systems across the USA.
* Soil types
ÿ­ Suitability/adaptability to varied soil types, including disturbed and marginal soils
* Establishment practices
ÿ­ Conventional tillage vs. no-till establishment
ÿ­ Planting date ÿ¿ variety interactions
ÿ­ Planting rates (and depths) appropriate for fiber and seed crops
ÿ­ Row spacing
* Fertilization practices
ÿ­ Application rates
ÿ­ Application timing (especially relative to different production outcomes (grain, fiber, dual purpose (grain + fiber), and essential oil production systems)
* Canopy management
ÿ­ Utility/timing of topping during growing season to induce multiple tillering
* Water use and demand
ÿ­ Irrigation ÿ¿ variety interactions
ÿ­ Evapotranspiration and water demand
ÿ­ Timing availability effects
* Insect, pathogens, and other pest management.
ÿ­ Pest and pathogen sensitivity
ÿ­ Efficacy of seed fungicide and insecticide treatments
ÿ­ Effects of late-season fungicide applications on grain yield and quality
ÿ­ Economic thresholds for insect and other pest control
* Weed management
ÿ­ Pre/post emergence herbicides for weed control
ÿ­ Herbicide sensitivities
* Harvest and handling practices
ÿ­ Evaluate efficacy of field desiccation (e.g., using diquat or glyphosate) for grain crops
ÿ­ Develop best practice protocols for retting both in fiber and dual purpose hemp systems
ÿ­ Determine engineering needs for harvest, handling and processing
* Suitability for crop rotations
ÿ­ Evaluate potential for use in rotations or mixtures with other crop
ÿ­ Measure hemp performance and weed/insect/disease incidence following corn, cotton, soybean, tobacco, forage/pasture/range/fallow
ÿ­ Determine hempâ¿¿s effects on disease/pest cycles of other crops
Project Methods
The 2018 Farm Bill fostered the introduction of hemp and many hemp-derived products into the food stream. As hemp is produced, harvested and dried there is the potential that pesticides may be applied intentionally or inadvertently through drift or cross-contamination. This raises the concern that pesticide residues may be found in hemp and/or hemp derived products that are intended for human use or consumption such as cannabidiol (CBD), seed and seed oil.The Connecticut Agricultural Experiment Station (CAES) has a long history of testing pesticides in food in Connecticut (CT). It has worked with the CT Department of Consumer Protection (CT DCP) since the 1963 and is the de facto laboratory to provide regulatory enforcement of pesticide residues in food sold within the state. Owing to the 2011 Food Safety Modernization Act (FSMA) the CAES attained accreditation to the International Organization for Standardization (ISO) 17025 standard in 2016 for the analysis of pesticide residues in foods.The CAES has experience employing the "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) method which is based on an acetonitrile/water partition and employs a dispersive solid phase extraction cleanup prior to analysis by gas and liquid chromatography with mass spectral detection (GC/MS) and (LC/MS). A total of 349 pesticide analytes (196 LC; 190 GC) have been validated in 30 different commodities / matrices (3 commodity groups; high water content; high acid and high-water content; high oil and intermediate water content) as part of our ISO 17025 accreditation.Given the increased amount of hemp and hemp-derived products in the marketplace, the concern over potential contamination of those products, and the experience of the CAES in analyzing these compounds, it is logical a validated method for these analytes in hemp be developed.The QuEChERS based approach used at the CAES will be applied to hemp bud, leaf and seed and / or hemp derived products. Once the method has been appropriately investigated, a single laboratory validation of the method will be performed working within the ISO 17025 guidelines. The detection limits, accuracy, reproducibility, and ruggedness of the method will be examined.To determine how prevalent pesticide residues might be in hemp grown in CT, largely as a result of potential spray drift, samples of hemp brought in from growers for delta-9 tetrahydrocannabinol (THC) testing can be tested for pesticide residues. Many hemp growers in CT currently submit samples to the CAES for the analysis of THC prior to harvest as required by the CT Department of Agriculture (CT DoAg).Hemp production is under study by the CAES at our Lockwood farm. A plot of hemp was planted in 2019, and again in 2020 to study various aspects of growth and production. This plot provides a unique opportunity to apply pesticides and allow weathering in the field to occur prior to harvest and analysis. This approach allows for the examination of field incurred pesticide residues. In addition it servesas a source of hemp for investigational laboratory studies. The test plot at the Lockwood farm also serves as an outreach and educational tool to the public and hemp growers. The CAES hosts an annual Plant Science Day (PSD) event. The CAES invites CT residents and interested parties to the farm to observe the field plots and to interact with scientists.Hemp growers in CT have primarily focused on the production of cannabidiol (CBD) owing to its high value as a commodity crop. Hemp derived cannabidiol (CBD) is in demand for its potential role in easing symptoms of many common health issues, including anxiety, depression, acne and heart disease. Pesticide residues may adulterate food by being carried along through the CBD extraction process from the hemp plant. As such it may be incorporated into oils and foods intended for human consumption.The fate of pesticide residues in the first step of most purification processes of CBD which is an ethanolic extraction will be examined. Either field or laboratory incurred residues on hemp will be extracted with ethanol at various temperatures and conditions. These extracts willand analyzed by our laboratory validated method for pesticide residues. The information obtained from these analysis will be valuable in determining potential health implications for pesticides in hemp derived product.After establishing the efficiency at which various pesticide residues are extracted into ethanol, the project can move forward, applying the QuEChERS based analysis method to commercially available CBD products for pesticide residue analysis. These efforts will help lead to a clearer understanding of how effectively pesticide residues on hemp may end up in products ingested by the consumer.

Progress 06/04/20 to 09/30/20

Outputs
Target Audience:The test plot of hemp grown by the Connecticut Agricultural Experiment Station (CAES) at our Lockwood farm in Hamden Connecticut (CT) serves as an outreach and educational tool to the public and growers. It provides seasonal summer students and other staff the opportunity to gain hands on knowledge in the growth, culling, harvest and production of hemp. It is further anticipated that the results and findings of this work will be shared through presentations at various formal and informal meetings with the public, local and federal agencies, and hemp growers. It will be shared with the scientific community and the general public through anticipated publications and or bulletins describing the findings.The CAES provides regulatory work in support of the CT Department of Agriculture (CT DoAg) and local hemp growers for delta-9 tetrahydrocannabinol (THC) testing. Hemp growers and the CT DoAg deliver samples to the CAES for testing. This provides additional opportunity for scientists to discuss issues related to hemp growing with growers and CT DoAg staff. Data and information obtained from this work will be directly shared CT DoAg which is responsible for the oversight of hemp production in CT as a raw agricultural commodity. Changes/Problems:During the timeframe in question our gas chromatograph with triple quadrupole detection (GC/MS) was inoperable. It was very recently repaired, and a new, replacement, instrument purchased. The Department of Analytical Chemistry (DAC) at the CAES received accreditation to the ISO/IEC 17025:2005 standard in December of 2016 from the American Association for Laboratory Accreditation (A2LA). The DAC was accredited 1) for the chemical testing of pesticide residues in food by gas- and liquid- chromatography with mass spectrometry (GC/MS and LC/MS) and 2) for arsenic in juice by inductively coupled plasma mass spectrometry (ICP-MS). In February 2018, the DAC was accredited to the updated ISO/IEC 17025:2017 standard. Three new items were included: 1) Testing pesticide residues using an alternative Liquid Chromatography High Resolution Mass Spectrometer (LC/HRMS), 2) Testing of solid and semi solid food matrices for arsenic by ICP-MS, and 3) the analysis of aflatoxin by Ultra High Pressure Liquid Chromatography with Mass Spectrometry (UHPLC/MS). In January 2021, the DAC will be involved in a scope expansion audit by A2LA to add the following to our scope of accreditation: 1) Total Delta-9 tetrahydrocannabinol (THC) and Cannabidiol (CBD) analysis in Hemp using GC-FID, 2) Percent crude Protein Analysis in animal feed by combustion method and 3) Percent crude Fat analysis in animal feed by Fat Extractor. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?The results of this work have been communicated to the hemp growing community in Connecticut (CT) through two separate webinars coordinated by The Connecticut Department of Agriculture (DOAG). What do you plan to do during the next reporting period to accomplish the goals?Our cloned hemp variety (Spec7) will be propagated in the Spring and grown in 2021 at the Lockwood farm in Hamden. This will provide continuity in the hemp bud that is used ongoing validation and extraction work. When the pesticide analytes are sufficiently validated by both gas and liquid chromatography with mass spectral detection (GC/MS) and (LC/MS) through spike-recovery work using frozen bud, we plan to move forward to examine the effect that processing hemp has on pesticide residues (i.e. ethanolic extraction). We can start this work with laboratory spikes, and plan to move forward to field spike, or to simulate those in the lab or greenhouse.

Impacts
What was accomplished under these goals? Eight feminized seed varieties were germinated and grown at the Lockwood farm. The varieties chosen were Abacus Early Bird, Cherry Abacus, Abacus 2.0, Abacus Early Bird 2.0, Cherry Abacus 2.0, Spec 7, and non-feminized seed from 2019 - Youngsim10 lot "2018WFS17A, Youngsim10 lot "2018WFS1." The field was planted with seedlings on June 25, 2020. Hemp buds from all varieties were collected and tested for delta-9 tetrahydrocannabinol (THC) and Cannabidiol (CBD) weekly by GC-FID from August 16 to October 26. Maximum CBD and THC concentration in the buds was found in all varieties during the week of September 19-26 (84-91days after planting). Hemp buds were harvested from the plants during this time frame. The buds have been stored frozen and will be used to obtain spike recovery data for pesticides of interest and to conduct initial CBD extraction work. On September 28, 2020 cuttings were taken from a single plant of flowering Spec7 Variety. These cuttings were successfully rooted and will provide a source for cloned hemp for the 2021 growing season. A sample of fresh bud that was harvested on September 28 was macerated then spiked in duplicate with our mix of 400 pesticides. The samples were extracted using the "quick, easy, cheap, effective, rugged, and safe" QuEChERS method which is under our ISO 17025:2017 scope of accreditation. The samples were analyzed by liquid chromatography with mass spectral detection (LCMS). Of the 196 analytes that are identified by LCMS, there were 159 that met our criteria for matrix expansion. This criterion is an average recovery from separate spikes of between of 50 and 150 percent. Although the hemp was spiked with 190 analytes that are detected by gas chromatography (GC) we were unable to perform this analysis because our GCMS inoperable for several months.

Publications