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Reﬁning Cannabidiol Using Wiped-Film Molecular Distillation:

Experimentation, Process Modeling, and Prediction
Article in Industrial & Engineering Chemistry Research · May 2022

DOI: 10.1021/acs.iecr.2c00290
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Refining cannabidiol (CBD) using wiped-film molecular

distillation: experimentation, process modeling, and
prediction
Journal: Industrial & Engineering Chemistry Research
Manuscript ID ie-2022-002906.R2
Manuscript Type: Article
Date Submitted by the

n/a
Author:
Complete List of Authors: Valizadehderakhshan, Mehrab; North Carolina Agricultural and Technical
State University, Engineering
Kazem-Rostami, Masoud; Macquarie University Department of Chemistry
and Biomolecular Sciences, Department of Molecular Sciences
Shahbazi, Abolghasem ; North Carolina Agricultural and Technical State
University
Azami, Mahsa; UNCG, Nanoscience
Bhowmik, Arnab; North Carolina Agricultural and Technical State
University
Wang, Lijun; North Carolina Agricultural and Technical State University,
Biological Engineering
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Refining cannabidiol (CBD) using wiped-film
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molecular distillation: experimentation, process
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modeling, and prediction
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16 Mehrab Valizadehderakhshan a, Masoud Kazem-Rostami b, c, Abolghasem Shahbazi d, *, Mahsa
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20 Azami e, Arnab Bhowmik d, Lijun Wang d
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24 a Joint School of Nanoscience and Nanoengineering, North Carolina Agricultural and Technical
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State University, Greensboro, NC 27401, USA
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b Department of Chemistry, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208,
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c Faculty of Science and Engineering, Macquarie University, North Ryde, NSW 2109, Australia
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50 * Corresponding author: Abolghasem Shahbazi, Biological Engineering Program, Natural Resources and
51 Environmental Design, North Carolina Agricultural and Technical State University, 1601 E. Market Street,
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53 Greensboro, NC 27411, United States. ash@ncat.edu; Tel.: +1-336-285-3830; fax: +1-336-334-7270
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d Natural Resources and Environmental Design, North Carolina Agricultural and Technical State
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7 University, 1601 E. Market Street, Greensboro, NC 27411, USA
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11 e Department of Nanoscience, Joint School of Nanoscience and Nanoengineering (JSNN),
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15 University of North Carolina at Greensboro, Greensboro, NC 27401, USA.
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4 ABSTRACT
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8 This work describes the stripping and refining of cannabidiol (CBD) from hemp extracts using a
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11 wiped film molecular distillation (WFMD) system. The process takes place in two stages, where
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the CBD gets stripped in the first stage and refined in the second stage. The main feed
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18 encompassing decarboxylated hemp extracts enters the stripping step at CBD concentration of
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21 about 35.8 wt.% and leaves at 48.0 wt.%, where the majority of terpenes leave the extracts. In
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refining stage the effects of process conditions, including pressure, evaporation temperature, and
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28 condensation temperature were examined using the response surface methodology (RSM)
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31 towards maximum CBD concentration and recovery. A second-stage recovery of 92.66 wt.%
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35 was achieved at the concentration of 80.19% by applying pressure of 40Pa, an evaporation
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38 temperature of 170 °C, and an internal condenser temperature of 20 °C. Analysis of the product
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41 showed that a low-pressure operation did not remove tetrahydrocannabinol (THC) from the
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45 CBD-rich product, thus proving to be an improper choice of WFMD for removing the
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48 psychoactive component. It was also found that the pressure reduction and increases in
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51 evaporation and condensation temperatures were contributing to higher CBD concentrations,
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4 although the condensation temperature had no effect on the recovery amount. RSM and artificial
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7 neural network (ANN) were further tested to assess their prediction capacity towards efficiency
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and process performance. It was found that both models offer satisfying prediction capability,
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14 although RSM had larger margins of error.
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4 1 Introduction
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6 Over 120 terpene derivatives and analogs 1, 90 cannabinoids 2, 20 flavonoids 3, and 16 other
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9 categories 1 account for over 750 identified natural chemicals 4 found in Cannabis sativa L.
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(hemp) 5. Among various plants, hemp is known to have the highest quantity of cannabinoids,
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16 which have immunologic effects on the human body. The antibacterial 6, anti-inflammatory 7,8,
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19 and antioxidant 9,10 properties of cannabinoids have been well-studied to date. However, the
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psychoactive effects associated with the existence of tetrahydrocannabinol (THC) turned out to
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26 be a major concern 11,12, resulting in hemp being classified as a controlled substance post World
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29 War II 13. More recently, public awareness of the medical benefits, technological options and
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33 economic aspects has drawn significant attention to cannabis and led to the push towards
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36 constitutional amendments in several countries 13,14. These developments with the growing
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39 interest in the recreational use of legal cannabis opened an unprecedented market for these
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43 materials. Today, the global hemp market is dominated mainly by one of its derivatives,
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46 cannabidiol (CBD) (51%), which stands above other hemp products, such as textiles (32%) and
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49 food (17%) 15.
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4 Nevertheless, the extraction, separation, and purification technologies have not been able to keep
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7 pace with the rapidly growing and ever eager market 16,17. For example, Rosenthal E. (2014)
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reported that only 40 – 70% of potential CBD is successfully obtained as a product, and the rest
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14 is lost during different steps of its refinement 4. The reasons behind the poor progress in the field
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17 are twofold: First, the limits imposed by legal impediments, and second, the lack of
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thermodynamic data as a critical barrier against modeling studies and equipment design and
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24 development 18,19. Among different separation technologies, vacuum evaporation and distillation
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27 offer several benefits, making it the proper choice for processing thermolabile blends. It takes
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advantage of vapor pressure and enables the boiling of different constituents at decreased
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34 temperatures. Various designs have been proposed to date, including falling-film 20 and wiped-
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37 film systems 21,22, which have been used in food, pharmaceutical, perfume, oil refining, and other
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41 chemical industries 19,23–25. A study of orange juice concentration by Chawankul et al. (2001)
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44 using an agitated thin film evaporator (ATFE) proved that the wiped-film evaporator apparatus is
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47 efficient in removing water from heat-sensitive food products 25. Eryzal et al. (2020)
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4 demonstrated the commercial potential of thin-film evaporators by looking at the economic
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7 benefits of their application in refining used lubricant oils 26.
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11 Although the aforementioned methods effectively reduce the boiling point and allow a high-
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throughput energy transfer, they are still unable to address a number of technical issues,
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18 including hot spot formation, lower yield, and processing of azeotropic mixtures. The alternative
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21 design, namely wiped-film molecular distillation (WFMD), incorporates both the heating and
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cooling surfaces into one chamber with a short molecule travel path 27–29. WFMD offers several
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28 unique advantages, including (i) superior energy transfer and continuous turbulence, resulting in
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31 a significant reduction in inner wall temperature requirements, (ii) maximized surface area per
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35 unit volume of flow, which facilitates evaporation, (iii) uniform distribution and exposure, thus
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38 preventing hot spot formation and material degradation, and (iv) enabling the processing of
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41 highly viscous materials. Additionally, as it is tailored to the input materials, tunable wiper
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45 speeds accommodate regulating the required temperature and pressure levels that allow for better
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48 separation 30.
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4 Cannabinoids and terpenes have relatively high boiling points and viscosities 31,32. While
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7 extracting cannabinoids and terpenes from hemp, usually a considerable quantity of undesirable
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co-extract species (including pigments, minerals, phospholipids, various types of herbal
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14 biopolymers, and other thermolabile components – collectively known as wax) 33, hurdle the
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17 post-processing steps. Despite the existing hints of benchtop use of WFMD—and sporadic
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mentions of it in commercial ads—no use of it has been found to this date in the literature about
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24 the refinement of cannabinoids and terpenes with a scientific approach on the evaluation and
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27 process optimization as a whole.
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31 This study aims to present a reliable approach to refine CBD using WFMD. In the spectrum of
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35 hemp extracts, CBD by far remains the dominant ingredient, and in terms of vapor pressure, it
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38 lays in the middle, nearly as the average 31,32. The governing thermodynamics of the extract
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41 mixture 10 demands a two-step operation procedure that strips CBD first, then enriches/purifies it
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45 next. This is mainly due to the difference in the vapor pressure of the constituents involved in the
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48 stripping and refinement steps of CBD. The alternative method for WFMD is short-path
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51 distillation. Short-path distillation is cost-effective and simple but suffers from low recovery and
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4 concentration 34. In this work, we examined the WFMD because it is fast and yields better
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7 recovery and concentration while operating at large scales. Crystallization technique can also
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further purify the final CBD product towards absolute purity; however, it is considerably slow
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14 and is limited scale-wise.
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18 In this study, we aim to achieve the following objectives: (1) introduce and evaluate a two-step
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21 distillation procedure for effective separation of terpenes and cannabinoids from each other, and
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heavy wax, (2) optimize operation parameters, aiming for a maximized CBD concentration and
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28 recovery, and (3) conduct a comparative study on process optimization and performance
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31 prediction between response surface method (RSM) and artificial neural network (ANN).
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2 Materials and methods
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39 2.1 Materials
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The hemp biomass used in this project was cultivated at North Carolina Agricultural and
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44 Technical State University (NCATSU). Once cropped, the flowers were separated and dried in
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47 the absence of sunlight at room temperature. Thereafter, the biomass was ground into powder
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4 using a 2mm mesh grinder. The powder moisture content was checked to make sure it did not
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7 exceed 11 wt.%.
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11 2.2 Cannabinoid extracts analysis
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13 Cannabinoid extracts were analyzed by Agilent’s Infinity-1260SII high-performance liquid
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16 chromatography (HPLC) instrument equipped with Waters C18 column (110Å, 5µm, 250 X
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20 4.6mm), and coupled with Agilent’s 6130 Quadrupole mass analyzer. HPLC chromatograms were
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23 recorded by setting the diode-array detector (DAD) signal channels at 210, 230, 250, and 280nm,
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26 with 4nm bandwidth referencing at 450nm with 100nm bandwidth at 25°C, using H2O/CH3CN
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30 mobile phase gradient from 10 to 90% organic over 20 min, which was followed by an isocratic
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33 elution of 90% organic (1.0 mL min–1 for 10 min). Solutions of cannabis isolates were
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36 microfiltered (0.2 µm, PTFE) before injections (2µL of 1 mg mL–1 in CH3CN) and analyzed by
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40 DAD to determine the time of retention tR for each component and to record their UV-Vis
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43 absorption spectra in the range of 200 – 400nm. A formic acid ammonium format (0.05 %) buffer
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solution was used for the mass detection running on electrospray ionization (ESI) at positive scan
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50 mode, resulting in the detection of 315.3 m/z (AR 100%) and 316.3 m/z (AR 21%) by single
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4 quadrupole MS within the positive ESI scan range of 100-500 Daltons, confirming the chemical
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7 formula of C21H31O2+ that stands for [CBD + H]+ 315.2318 m/z.
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11 2.3 Nuclear magnetic resonance (NMR)
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13 The isolated fractions were dissolved in deuterated chloroform (CDCl3), and their proton spectra
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16 were recorded by Varian INOVA-600MHz NMR spectrometer at a controlled ambient temperature
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20 at 300K. The obtained FIDs were processed by Topspin V.4 and MestReNova V.14. Every NMR
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23 spectrum obtained from the CDCl3 solution was calibrated for 1H of the residual non-deuterated
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26 solvent at 7.26 ppm. A commercially available CBD isolate was purchased from BestLife™ and
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30 analyzed as a reference to facilitate the comparative NMR studies. The NMR spectra and the
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33 assignment of corresponding peaks are included in Figure S1 of the Supporting Information (SI).
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37 2.4 Experimental
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39 2.4.1 Extraction
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41 Prior to large-scale extractions, a screening study was conducted using isopropanol, methanol,
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44 ethanol, and hexane to compare their extraction performances. All solvents were analytical grade
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purchased from Merck® and were used as received. Figure S2 shows biomass extraction (wt.%)
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51 against immersion time for each solvent used. The preliminary extraction yield confirmed the
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4 superior performance of ethanol and isopropanol over others in terms of time and yield. Finally,
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7 ethanol was chosen for large-scale extractions due to its low toxicity and ease of recovery. A
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large-scale ethanol extraction (temperatures < –27°C) was performed in batches using Precision®
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14 Extraction Solutions - Model X10 MSE setup where the biomass ethanol mass ratio was set to
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17 1:3 (see Figure S3 in the SI).
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21 2.4.2 Winterization
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23 Wax is an inhomogeneous extract component that includes complex structures and tends to
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26 denature at elevated temperatures. Post-extraction winterization (intense freezing) results in
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effective wax removal by solubility reduction 4. The remaining extract was re-dissolved in
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33 ethanol (extract to solvent ratio of 1:10 w/w) and was chilled and kept at T< –70 °C for 24h
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36 before draining the solid sediments. The remaining solvent was rotary-evaporated, leaving a
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40 brownish gooey sticky extract.
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44 2.4.3 Decarboxylation
45 Based on Le-Chatelier’s principal, WFMD meets the favorable conditions for the endothermal
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49 decarboxylation reaction (low pressure and exerting sufficient thermal energy), hence making it
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52 inevitable. On the other hand, while operating under low pressure, the release of CO2 via
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4 decarboxylation causes pressure instabilities and other technical issues. To mitigate these, the
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before subjecting it to distillation. The heat-treated winterized extract was then transferred to the
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14 WFMD feed tank and degassed under vacuum pressure.
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18 2.4.4 Experimental setup
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A 2-inches Short-Path Still WFMD manufactured by POPE™ Inc., USA, performed the
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23 distillation, providing an effective evaporation surface area of 0.0325 m2. The WFMD machine
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26 used to conduct the distillation is shown in Figure 1. An electric element installed outside the
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main column surrounded by a thermal jacket supplied the required heat. The wiper (three-
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33 paddled) speed and feed flowrate were kept at 360 rpm and an under-design rate of 100 ml.hr-1,
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36 respectively, assuring that the film thickness was not the limiting factor to the mass transfer.
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40 Collected samples were stored at 0°C before the subsequent analysis.
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44 Because the WFMD is a once-through system with constant pressure throughout the system,
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47 stripping and refinement of CBD are not feasible in one stage; therefore, a double-pass process
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50 was necessary. CBD stripping (the first stage) involves the degassing and removal of the
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4 components with vapor pressures higher than those of cannabinoids (e.g., solvents and most of
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7 the terpenes), and therefore, the obtained stream was named terpenes (for simplification). The
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second step (CBD refinement) mainly separates a cannabinoid-rich stream from the non-
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14 evaporating residuals, referred to as wax. The CBD concentration and recovery changes were
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17 monitored and analyzed by using NMR, HPLC, and MS instruments. Table 1 summarizes the
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employed parameters used in each operational step.
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Table 1. The operation parameters used in each step
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28 Inlet Wiper Pressure Evaporator Internal condensation External Moisture
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30 flow speed temperature temperature condensation trap
31 (P) (EvapT) (CondT) temperature temperature
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33 First 100 360 666 Pa 140 °C –15 °C –20 °C –60 °C
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35 step mL h-1 RPM
36 Second 100 360 Subjected to optimization study detailed in –20 °C –60 °C
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38 step mL h-1 RPM the following section
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37 Figure 1. Wiped-film molecular distillation (WFMD) machine
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4 2.4.5 Experiment design
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Optimization of CBD concentration ([CBD]) and CBD recovery were considered by examining
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9 the affecting factors. CBD recovery is the gain of CBD within the final product stream and can
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12 be defined as (Eq. 1) 27:
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17 (𝑚𝑙ℎ𝑟) × [𝐶𝐵𝐷]
𝑃𝑟𝑜𝑑 𝑓𝑙𝑜𝑤 𝑝𝑟𝑜𝑑
18 𝐶𝐵𝐷 𝑅𝑒𝑐𝑜𝑣𝑒𝑟𝑦 (%) = × 100 (Eq. 1)
𝐹𝑒𝑒𝑑 𝑓𝑙𝑜𝑤 ( ) × [𝐶𝐵𝐷]
19 𝑚𝑙
20 𝑓𝑒𝑒𝑑
ℎ𝑟
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24 Unlike the seemingly linear relation between the recovery (%) and [CBD], the increase in
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27 [CBD]prod usually costs in the decrease of product flow. To optimize these parameters, three
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30 independent factors, including the operation pressure (P in Pa), evaporator temperature (EvapT
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34 in °C), and internal condenser temperature (CondT in °C), each with three levels, were
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37 considered. Compared to 33 full-factorial, the Box-Behnken design (BBD) has been proven to be
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40 economical and efficient 35. Therefore, BBD was adopted, and 17 experiments were designed
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44 using DesignExpert7.0.0.
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48 The data from analysis of experiments were collected, analyzed, and fit into a second-order
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51 polynomial (Eq. 2), followed by an analysis of variance (ANOVA):
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1
2
3 3 3 2 3
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𝛶 = 𝛽0 + ∑𝛽 𝑋 + ∑𝛽
𝑖=1
𝑖 𝑖
𝑖=1
𝑖𝑖 𝑋2𝑖 + ∑ ∑ 𝛽 𝑋𝑋
𝑖 = 1𝑗 = 𝑖 + 1
𝑖𝑗 𝑖 𝑗 (Eq. 2)
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9 Where Y is the response; β0, βi, βii, βij are the regression coefficients estimated using the equation
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12 for the intercept, linearity, square, and interaction, respectively; and Xi and Xj represent the
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independent variables (X1 = pressure, X2 = evaporator temperature, X3 = internal condenser
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19 temperature).
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23 2.4.6 WFMD modeling using artificial neural networks (ANN)
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25 In addition to the response surface method (RSM), ANN was also deployed to predict the
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28 response values. ANN is a class of linear modeling techniques that learns the general behavior of
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31 a process and then predicts a potential outcome using its trained neurons. The procedure involves
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35 the selection of network architecture, determination of hidden layers and number of neurons in
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38 each layer, learning, training, validation, and verification of the data 36. In this study, the three
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41 independent variables, namely P, EvapT, and CondT, were considered input neurons. CBD
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45 concentration ([CBD]) and CBD recovery were also considered the output neurons in separate
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48 studies, where each network carries only one response. Figure 2 schematically shows the feed-
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forward Multilayer Perceptron (MLP) model containing two neurons in its hidden layer. The
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4 process was simulated using the Neural Net Fitting toolbox in MATLAB R2021a. The MLP was
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This algorithm is typically fast and stops once the generalization stops improving. This
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14 procedure minimizes squared errors as the training objectives function and produces a network
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17 that generalizes well 37. The hyperbolic tangent sigmoid transfer function (tansig) was applied in
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both hidden and output layers.
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39
40
41
42 Figure 2. Structure of the neural network used in the WFMD modeling.
43
44
45
ANN and RSM were compared in terms of their performance and their error margins. To do so,
46
47
48
49 the mean absolute error (MAE), mean absolute percentage error (MAPE), root mean square error
50
51
52
53
54
55
56 18
57
58
59
1
2
3
4 (RMSE), and the correlation coefficient (R2) were calculated (Eq. 3 – (Eq. 6) and considered in
5
6
7 examining the ability of each method in process prediction 36,38.
8
9
10 𝑁
11 1
12
13
𝑀𝐴𝐸 =
𝑁 ∑|𝑌
𝑖=1
𝑒𝑥𝑝𝑖 ― 𝑌𝑝𝑟𝑒𝑑𝑖| (Eq. 3)
14
15
16 100 𝑁 |𝑌𝑒𝑥𝑝𝑖 ― 𝑌𝑝𝑟𝑒𝑑𝑖|
17 𝑀𝐴𝑃𝐸 =
𝑁 ∑ 𝑖=1
(
𝑌𝑒𝑥𝑝𝑖
) (Eq. 4)
18
19
20 𝑁
21 1
22 𝑅𝑀𝑆𝐸 =
𝑁 ∑(𝑌 𝑒𝑥𝑝𝑖 ― 𝑌𝑝𝑟𝑒𝑑𝑖)2 (Eq. 5)
23 𝑖=1
24
25 𝑁
26 (𝑌𝑒𝑥𝑝𝑖 ― 𝑌𝑝𝑟𝑒𝑑𝑖)2
27
2
𝑅 = 1― ∑ 2 (Eq. 6)
28
29
30
𝑖=1
[𝑌𝑒𝑥𝑝𝑖 ―
1 𝑁
()
∑ (𝑌 )]
𝑁 𝑖 = 1 𝑒𝑥𝑝𝑖
31
32 Here, N is the number of experiments, Yexp is the experimental data, and Ypred is the
33
34
35
36
corresponding Y value predicted either by ANN or RSM.
37
38
39
40
41 3 Results and discussions
42
43 3.1 Process overview
44
45 The operating parametric values in the first step (Table 1) are selected such that the available
46
47
48 CBD in the main feed gets stripped i.e., the majority of the lighter components leave the blend.
49
50
51
52
During the stripping (first) stage, almost 25% of the main feed ([CBD] = 35.87wt.%) goes to the
53
54
55
56 19
57
58
59
1
2
3
4 terpene stream at the CBD concentration of about 11.6%. The distillate bottom conveying the
5
6
7 majority of CBD carries almost 73% of the main input flow. The stripped blend is subsequently
8
9
10
subjected to more aggressive conditions during the second step for purification. The second-step
11
12
13
14 feed (2nd feed) with a CBD concentration of 45.28% was fed at the rate of 100 mL h-1 under
15
16
17 different operation conditions to refine the distillate stream. Table 2 represents the Box-Behnken
18
19
20
21
design matrix of the second step at which the lower, central, and upper levels were demonstrated
22
23
24 by (–1), (0), and (+1), respectively. During each experiment, the collected mass from the internal
25
26
27 condenser (CBD-rich stream, known as product) was sampled and analyzed for CBD and THC.
28
29
30 𝑃𝑟𝑜𝑑
31
Using product mass flow, the (2𝑛𝑑 𝐹𝑒𝑒𝑑) ratio, and thereby the CBD recovery was calculated using
32
33
34 (Eq. 1). The sample obtained from a test carried out under the highest evaporation temperature
35
36
37
38
and vacuum was named HTV.
39
40
41
42
Table 2. Experimental parameters during the second step of WFMD
43
44
45 Operation parameters 2nd step product characterization
46 𝐶𝐵𝐷
47 P EvapT CondT Approx. [CBD]* [THC]* 𝑇𝐻𝐶 CBD recovery
𝑃𝑟𝑜𝑑
48 (Pa) (°C) (°C) (2𝑛𝑑 𝐹𝑒𝑒𝑑)% (wt.%) (wt.%) (wt.%)
49
50 1 40 (–1) 110 (–1) 20 (0) 43 71.51 2.30 31.09 68.33
51
52 2(HTV) 40 (–1) 170 (+1) 20 (0) 52 80.19 2.76 29.05 92.66
53
54
55
56 20
57
58
59
1
2
3
4 3 40 (–1) 140 (0) –10 (–1) 50 73.28 2.37 30.92 81.42
5
4 40 (–1) 140 (0) 50 (+1) 46 77.63 2.52 30.81 79.36
6
7 5 220 (0) 140 (0) 20 (0) 40 60.72 1.94 31.30 53.97
8
9 6 220 (0) 140 (0) 20 (0) 32 52.86 1.76 30.03 37.59
10
7 220 (0) 140 (0) 20 (0) 36 57.97 1.67 34.71 46.38
11
12 8 220 (0) 140 (0) 20 (0) 33 59.88 1.82 32.90 43.91
13
14 9 220 (0) 140 (0) 20 (0) 42 58.15 1.86 31.26 54.27
15
10 220 (0) 110 (–1) –10 (–1) 21 45.63 1.34 34.05 21.29
16
17 11 220 (0) 170 (+1) –10 (–1) 35 64.76 2.10 30.84 50.37
18
19 12 220 (0) 110 (–1) 50 (+1) 33 48.54 1.48 32.80 35.60
20
21
13 220 (0) 170 (+1) 50 (+1) 38 68.42 2.48 27.59 57.78
22 14 400 (+1) 110 (–1) 20 (0) 15 30.76 0.97 31.71 10.25
23
24 15 400 (+1) 170 (+1) 20 (0) 21 49.87 1.70 29.34 23.27
25
26
16 400 (+1) 140 (0) –10 (–1) 17 38.58 1.23 31.37 14.57
27 17 400 (+1) 140 (0) 50 (+1) 16 44.51 1.41 31.57 15.83
28
29 *The values include the common thermal and oxidation artifacts of the decarboxylated form of
30
31 cannabinoids.
32
33
34
35
36
37 During the second step, applying the lowest evaporator temperature, the highest mass fraction of
38
39
40 wax was obtained 39. Inversely, with the increase of EvapT, as expected, a bigger portion of the
41
42
43 𝑃𝑟𝑜𝑑
44 inlet feed (the 2nd feed) transferred to distillate, hence a higher 2𝑛𝑑 𝐹𝑒𝑒𝑑 ratio was obtained.
45
46
47 Elevated EvapT may lead to a higher denaturing rate and solidification of the thermolabile
48
49
50
51
52
53
54
55
56 21
57
58
59
1
2
3
4 components of the feed—e.g., pigments, phospholipids, etc.—which at its extreme may clog the
5
6
7 exit elbow (Figure 1).
8
9
10
11 3.2 Product analysis
12
13 The original winterized and decarboxylated feed submitted to WFMD system was divided into
14
15
16 two main fractions, one constituted the first-step bottom outlet (about 73wt.%) and the other was
17
18
19
20 a terpene-rich stream distillate (25 wt.%). This process also ended with the accumulation of
21
22
23 volatiles (less than 2 wt.%) in the external condenser and cold trap that includes the solvent
24
25
26 residues and moisture (Figure 1). In the second step, however, the product characteristics,
27
28
29
30 recovery, and external condenser collections varied depending on the applied operating conditions
31
32
33 (detailed in Table 2).
34
35
36
37 Because of the employed intense conditions in HTV, this sample demonstrated to have the
38
39
40 𝑃𝑟𝑜𝑑
highest [CBD] and 2𝑛𝑑 𝐹𝑒𝑒𝑑 ratio, thus the highest CBD recovery (Eq. 1). Despite the profound
41
42
43 𝐶𝐵𝐷
44 changes in the operation conditions (Table 2) and, therefore, [CBD] changes, the 𝑇𝐻𝐶 ratio
45
46
47
remained almost the same, proving that employing different operational conditions did not
48
49
50
51 contribute to separating CBD from its psychoactive counterpart, THC. Table 3 summarizes the
52
53
54
55
56 22
57
58
59
1
2
3
4 analysis of the main streams involved in the double-step enrichment process. The quantity of
5
6
7 natural cannabinoids (CBDA and THCA) has been reduced to none in all the process streams,
8
9
10
implying a successful and thorough decarboxylation. Typically, when products containing
11
12
13
14 natural cannabinoids are subjected to thermophysical processes, they undergo potential
15
16
17 transformations that make them unsuitable for medical and analytical applications, as they may
18
19
20
21
lead to further production of the psychoactive agent.
22
23
24
25
Table 3. Cannabinoid analysis of major streams
26
27 𝐶𝐵𝐷
28 Sample CBD THC 𝑇𝐻𝐶 CBDA or
29
30 (wt.%) (wt.%) THCA
31 1st Feed 35.87 1.80 16.79 Trace
32
33 Terpene outlet 11.67 0.23 50.73 Non
34
35 2nd Feed 45.28 1.59 28.78 Non
36 HTV Wax 16.67 1.05 15.87 Non
37
38 HTV Product 80.19 2.76 29.05 Non
39
40
41
42
43
44 To further examine the success of stripping and purification stages, the product streams from the
45
46
47 first step (terpene-rich) and the second step (CBD-rich) were further analyzed using NMR to
48
49
50
trace the undetected components and monitor their qualitative purity. Figure 3 compares the
51
52
53
54
55
56 23
57
58
59
1
2
3
4 stacked 1H NMR spectra of pure CBD, HTV product, and terpene stream (1st step), numbered 1–
5
6
7 3, respectively.
8
9
10
11 According to the NMR spectrum of pure CBD (Figure 3, Spectra 1), the major component of
12
13
14
sample number 2 is indeed CBD, while sample 3 carries a significant quantity of impurities.
15
16
17
18 Based on the peak shape and the unique chemical shift of a singlet at 5.57 ppm, which was
19
20
21 assigned for one vinyl proton, it was chosen as the reference for the comparative NMR
22
23
24
25
integrations. This sharp singlet pattern reduces the chances for overlapping in this region of
26
27
28 chemical shift, which mainly belongs to vinyl groups and does not allocate a vast majority of
29
30
31 other functional groups. In addition, the absence of disadvantageous diastereotopic and coupling
32
33
34
35 effects increased the sharpness of this peak, making it the most suitable reference peak among
36
37
38 the rest. Based on the integrals of the assigned reference peaks for CBD, samples 2 and 3 showed
39
40
41 a relative purity percentage of 13.14 % and 85.94 %, respectively. These findings are in
42
43
44
45 agreement with the product analysis parallelly conducted via reversed-phase HPLC. The
46
47
48 comparative HPLC analysis of the final products (HTV-Wax and CBD-rich HTV product) and
49
50
51 pure CBD are shown in Figure S4 of the SI.
52
53
54
55
56 24
57
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59
1
2
3
4
5
6
7
8
9
10
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17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
Figure 3. 1H-NMR analysis of the major streams (600 MHz, CDCl3, 300K). The spectra represent
36
37
38 pure CBD (1), HTV product (2), and terpene-rich stream (3), respectively.
39
40
41
3.3 The effects of operating parameters on CBD concentration (2nd stage)
42
43 The applied temperature of 140 °C during the first stage (P= 666 Pa) removed the most volatile
44
45
46
47
terpenes leaving the cannabinoids and wax behind 40,41. The pressure-temperature nomograph
48
49
50 interactive tool 42 provides an approximate boiling profile of different chemicals under reduced
51
52
53
54
55
56 25
57
58
59
1
2
3
4 pressures. During the second step, the employed operating conditions cover a broad CBD
5
6
7 concentration ranging from 30 – 80 wt.% (Table 2). At the lower end of the scale, the product
8
9
10
possesses less [CBD] than the feed (45.28 wt.%), indicating that the employed conditions
11
12
13
14 stripped the desired component, i.e., the CBD was progressively stripped and lost from the
15
16
17 product stream. While operating in this condition, terpenes with low vapor pressure (e.g.,
18
19
20
21
terpineol, linalool, eucalyptol, and D-limonene) comprise most of the product stream collected in
22
23
24 the internal condenser flask. Conversely, with the increase of EvapT and/or decrease of operating
25
26
27 pressure, more CBD jumps through the short path to join the product stream, surpassing the feed
28
29
30
31
CBD concentration at P = 220 Pa, EvapT = 110 °C, and CondT = –10 °C.
32
33
34
35 RSM is a statistical tool that allows us to find the relationship between the response(s) and the
36
37
38 independent variables. This technique fits experimental data to the mathematical models to
39
40
41 provide a better picture of the general response behavior. RSM can be used for response
42
43
44
45 interpretation, prediction, and more importantly, enhance the efficiency of the experiments
46
47
48 35,36,43. In this study, RSM was used to investigate the effect of operation parameters on both
49
50
51 concentration and recovery. Figure 4 represents the response plots of [CBD] versus the
52
53
54
55
56 26
57
58
59
1
2
3
4 independent variables. After eliminating non-significant model terms, (Eq. 7 was obtained,
5
6
7 showing a semi-linear functionality of the product concentration versus the operating parameters.
8
9
10
The adequacy of the model was evaluated by ANOVA (Table 4). The model F-value of 144.58
11
12
13
14 implies the model is significant. There is only a 0.01% chance that a model F-value this large
15
16
17 could occur due to noise. Values of Prob > F less than 0.05 indicate that model terms are
18
19
20
21
significant (Eq. 7). The "Lack of Fit F-value" of 0.34 implies the lack of fit is not significant
22
23
24 relative to the pure error. (Eq. 7 applies to the pressure and evaporator temperature ranges
25
26
27 between 40Pa < P < 400Pa and 110°C < EvapT < 170°C, respectively.
28
29
30
31 Table 4. Analysis of variance of reduced quadratic model for [CBD] vs. affecting factors
32
33
34
35 Sum of p-value
36 Source Squares df Mean Square F- Value (Prob > F)
37
38 Model 3031.77 4 757.94 144.58 < 0.0001 significant
39
40 A – P (Pa) 2411.30 1 2411.30 459.96 < 0.0001
41 B – EvapT (°C)557.78 1 557.78 106.40 < 0.0001
42
43 C – CondT 35.49 1 35.49 6.77 0.0231
44
45 (°C)
46 AB 27.20 1 27.20 5.19 0.0419
47
48 Residual 62.91 12 5.24
49
50 Lack of Fit 25.57 8 3.20 0.34 0.9079 not significant
51
Pure Error 37.34 4 9.34
52
53
54
55
56 27
57
58
59
1
2
3
4 Cor Total 3094.68 16
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29 (a) (b)
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48 (c) (d)
49
50
51
52 Figure 4. Response surface and contour curves of CBD concentration.
53
54
55
56 28
57
58
59
1
2
3
4 Figure 4 (a) shows the greatest range (30 – 81%), indicating the significance of the factors.
5
6
7 Noticeably, the semi-parallel contours of CBD with CondT confirm that the CondT has provided
8
9
10
a minor contribution to the final product concentration (Figure 4 – b and c). It can be attributed to
11
12
13
14 two hypotheses, (1) the employed condenser range is lower than the dew point of CBD in the
15
16
17 operation pressure, and (2) the chemical of interest is a dominant component within the blend.
18
19
20
21
Both hypotheses hold true from the thermodynamic data of CBD and Table 2.
22
23
24
25 [𝐶𝐵𝐷] = +53.56–0.16 × 𝑃 + 0.17 × 𝐸𝑣𝑎𝑝𝑇 + 0.07 × 𝐶𝑜𝑛𝑑𝑇 + 4.83 × 10 ―4 × 𝑃 × 𝐸𝑣𝑎𝑝𝑇 (Eq
26
27 . 7)
28
29
30 The response contour lines and the plot slopes show that [CBD] is the greatest function of
31
32
33
34 pressure, which is also evident in the final model (Eq. 7). In determining the significant model
35
36
37 terms, the p-value was taken into account, then the non-significant model terms were omitted44,
38
39
40
and (Eq. 7) was obtained. This equation shows that the effects of pressure and evaporator
41
42
43
44 temperature are higher than that of condenser temperature. Additionally, as shown in Figure 4-d,
45
46
47 the interaction term in the model (Eq. 7) is still considered significant (p-value = 0.0419), which
48
49
50
51
justifies the marginal curvature in Figure 4-a. Similar results were reported by Dantas et al. 39
52
53
54
55
56 29
57
58
59
1
2
3
4 and Martines et al. 28, investigating the effect of different parameters on product concentration in
5
6
7 molecular distillation systems.
8
9
10
11 3.4 The effect of operating parameters on CBD recovery (the 2nd stage)
12
13 The selected factors demonstrated different influences on CBD recovery. These effects were
14
15
16 analyzed for ANOVA and statistical significance, considering a confidence level of 94% (chance
17
18
19
20 of a variable influencing a response). Table 5 is the ANOVA table evaluating the model terms of
21
22
23 CBD recovery, and the obtained model describing these effects is given by (Eq. 8). The model F-
24
25
26 value of 145.75 implies the model is significant. There is only a 0.01% chance that a model F-
27
28
29
30 value this large could occur due to noise. Only the pressure and the EvapT were found to be
31
32
33 significant (Prob > F). The working temperature of the internal condenser was found to be
34
35
36 statistically insignificant, i.e., of no substantial contribution to the recovery percentage (%).
37
38
39
40
Table 5. ANOVA table for reduced response surface of CBD recovery against affecting factors
41
42
43
44 Sum of p-value
45
Source Squares df Mean Square F- Value Prob > F
46
47 Model 9292.29 2 4646.15 123.63 < 0.0001 significant
48
49 A – P (Pa) 8310.83 1 8310.83 221.15 < 0.0001
50
51
B – EvapT (°C) 981.47 1 981.47 26.12 0.0002
52 Residual 526.13 14 37.58
53
54
55
56 30
57
58
59
1
2
3
4 Lack of Fit 326.46 10 32.65 0.65 0.7335 not significant
5
Pure Error 199.66 4 49.92
6
7 Cor Total 9818.42 16
8
9
10
11
12
13 Recovery (wt.%) = + 33.99 – 0.18 × P + 0.37 × EvapT (Eq. 8)
14
15
16 Figure 5 (a) demonstrates the response plot of the CBD recovery against the significant operating
17
18
19
20
parameters, and (b) schematically shows the effect of all parameters on the response around/near
21
22
23 their central point before truncating the model. The 2nd-stage recovery percentage starts at about
24
25
26 7wt.%—at 400Pa and evaporator temperature of 110 °C—reaching the all-time maximum of
27
28
29
30 nearly 93% in HTV conditions which Table 2 displays for the optimal double-stage recovery of
31
32
33 85.1%. The perturbation graph of the un-truncated model (Figure 5-b) clearly shows that the
34
35
36 effect of process pressure on CBD recovery is skyrocketing. The overall evaporation chamber
37
38
39
40 temperature rises as the internal condenser temperature increases, pushing the lighter
41
42
43 components out of the chamber that results in an increase in CBD concentration in the final
44
45
46 product. However, as previously stated, technical issues such as polymerization occur at higher
47
48
49
50 chamber temperatures, preventing further recovery. Therefore, as can also be seen in (Eq. 8), the
51
52
53
54
55
56 31
57
58
59
1
2
3
4 effect of CondT was found to be non-significant on CBD recovery. The pressure level and
5
6
7 evaporation temperature were the key parameters in this equation, with no interaction term of
8
9
10
statistical significance that could affect the overall recovery.
11
12
13
14
Reaching the CBD recovery of 93% from the 2nd feed is outstanding, as it means only 7% of the
15
16
17
18 CBD in the 2nd feed has not been successfully collected in the product stream. When working on
19
20
21 the effect of agitation speed, evaporator temperature, and condenser temperature on patchoulol
22
23
24
25
enrichment using molecular distillation, Dantas et al. (2020) found that the internal condenser
26
27
28 temperature exerted a weak influence on the product recovery 39. They reported no significant
29
30
31 interaction between the affecting parameters on the recovery ratio, which is in agreement with
32
33
34
35 our findings.
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56 32
57
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20 (a) (b)
21
22
23
24 Figure 5. CBD recovery (wt.%) vs operation parameters; response surface (a) and, Perturbation in
25
26
27
coded form (b). A: pressure, B: evaporator temperature, and C: internal condenser temperature.
28
29
30 Figure 6 illustrates the 3D presentation of the reversed-phase HPLC chromatogram for the
31
32
33 fraction obtained by HTV in which the time of retention (tR in minutes) is against the absorbed
34
35
36
37
wavelengths (λ in nanometers) against the intensity of the incident light at the given wavelengths
38
39
40 in thousandth of an absorbance unit (mAu). This example intentionally exceeds the Beer-
41
42
43 Lambert reliability zone for the major component due to the high concentration of the sample to
44
45
46
47 visualize the smaller peaks related to the negligible impurities. This indicates that CBD
48
49
50 dominates the sample as the only major component, as it is exceedingly evident from the
51
52
53
54
55
56 33
57
58
59
1
2
3
4 characteristic peak, retention time, the acquired mass spectra, and the UV absorption profile.
5
6
7 Nonetheless, NMR spectroscopy was parallelly conducted to determine whether the negligible
8
9
10
impurities are, in fact, minor in concentration and not transparent to UV light or invisible to the
11
12
13
14 DAD. The NMR experiments confirmed the only major component was indeed CBD, and those
15
16
17 negligible impurities did not exceed 15wt.% collectively.
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43 Figure 6. 3D plot of reversed-phase HPLC chromatogram of HTV sample obtained at P=40Pa,
44
45
46
EvapT=170 °C, and CondT = 50 °C, [CBD] = 80.19%
47
48
49
50
51
52
53
54
55
56 34
57
58
59
1
2
3
4 3.5 Model of artificial neural network
5
6
CBD concentration and recovery were simulated in separate studies, wherein in each study, 70%
7
8
9 of the experimental data were used to train the model. Successive model re-training was
10
11
12 conducted, and the best models were determined using the statistical indicators. The results of
13
14
15
16 ANN regression for training, testing, validation, and the general state for both responses are
17
18
19 shown in Figure 7.
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
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36
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56 35
57
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59
1
2
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7
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34
35 (a)
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56 36
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33 (b)
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37 Figure 7. ANN Regression graphs for training data, test data, validation data, and the overall
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40 model for CBD concentration (a) and recovery (b). The target implies the experimental data.
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43 From (Eq. 3 – (Eq. 6, depending on the distribution of datapoints, the employed ANN algorithm,
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47 as well as the value of N in each test (or the overall regression), the R-value can vary
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50 significantly. Overall, good correlation coefficients were obtained for all data simulated by both
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56 37
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4 ANN models, indicating that the models can efficiently optimize the process parameters. Table 6
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7 reports the predicted response values by ANN and RSM models along with the corresponding
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experimental values.
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Table 6. Process variables, experimental results, predicted values by ANN and RSM for CBD
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17 concentration and recovery.
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20 Operation parameters [CBD] wt.% CBD recovery wt.%
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22 # P EvapT CondT Exp. Predicted Exp. Predicted
23 (Pa) (°C) (°C) ANN RSM ANN RSM
24
1 40 110 20 71.51 68.99 69.39 68.33 69.19 67.49
25
26 2 (HTV) 40 170 20 80.19 81.02 80.74 92.66 87.46 89.69
27 3 40 140 -10 73.28 75.21 72.96 81.42 76.80 78.59
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29 4 40 140 50 77.63 77.61 77.16 79.36 81.04 78.59
30 5 220 140 20 60.72 58.23 58.44 53.97 44.61 46.19
31
6 220 140 20 52.86 58.23 58.44 37.59 44.61 46.19
32
33 7 220 140 20 57.97 58.23 58.44 46.38 44.61 46.19
34 8 220 140 20 59.88 58.23 58.44 43.91 44.61 46.19
35
36 9 220 140 20 58.15 58.23 58.44 54.27 44.61 46.19
37 10 220 110 -10 45.63 45.50 48.05 21.29 24.75 35.09
38
11 220 170 -10 64.76 64.95 64.62 50.37 53.02 57.29
39
40 12 220 110 50 48.54 50.60 52.25 35.60 36.33 35.09
41 13 220 170 50 68.42 67.48 68.82 57.78 66.86 57.29
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43 14 400 110 20 30.76 31.77 30.91 10.25 8.55 2.69
44 15 400 170 20 49.87 49.70 52.70 23.27 23.28 24.89
45
16 400 140 -10 38.58 37.15 39.71 14.57 14.63 13.79
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47 17 400 140 50 44.51 42.69 43.91 15.83 16.22 13.79
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56 38
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1
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4 The ANN prediction results for both outputs show close proximity to the actual performance of
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7 the process, confirming that the ANN model has an acceptable precision. Table 7 compares the
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ANN and RSM predictions by summarizing the statistical indicators from (Eq. 3 – (Eq. 6.
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Table 7. The statistical parameters used to compare ANN and RSM.
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18 [CBD] CBD recovery (%)
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ANN RSM ANN RSM
21 MAE 1.35 1.465 3.47 4.00
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23 MAPE 2.50 2.76 7.8 14.50
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25 RMSE 1.88 2.08 4.79 5.57
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27 R2 0.9807 0.9797 0.9605 0.9552
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From Table 7, it can be concluded that both models offer acceptable abilities to design and
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36 interpret process data; however, the ANN exhibited relative superiority in predicting the
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39 experimental results over RSM, both for [CBD] and CBD recovery 45. Even though the
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correlation coefficient of RSM for CBD recovery is still higher than 95%, the model is less
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46 reliable due to the extravagant higher error values, as can be seen from the obtained results in
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49 Table 6. This can be attributed to numerous RSM model terms for CBD recovery (Eq. 8), which
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53 have been crossed out due to insignificance, suggested by ANOVA analysis. Due to
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56 39
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4 experimental hurdles in the calculation of recovery values, the predictions offered by both
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7 models showed lesser accuracy than those to predict [CBD].
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4 Conclusion
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This work optimized the major parameters of WFMD to obtain CBD-rich products from hemp
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18 extracts, using response surface and artificial neural models. The Box-Behnken planning
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21 provided mathematical representations describing the CBD concentration and recovery vis-à-vis
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the considered parameters. Among different operational settings tested for refinement, the most
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28 optimum setting include pressure of 40Pa, evaporation temperature of 170 °C, and internal
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31 condenser temperature of 20 °C, resulting in a maximum CBD concentration of 80.19% and
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35 recovery of 92.66%. Within the tested parameters ranges, it was found that pressure reduction
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38 increases both the concentration and recovery of CBD, and this contribution is stronger than the
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41 effect of other parameters. It was also discovered that the WFMD system is not a proper
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45 apparatus for obtaining medical-grade products, as it did not help with the separation of CBD
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48 from psychoactive THC.
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56 40
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4 Both RSM and ANN were able to offer good predictions of the CBD concentration. In the case
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7 of recovery, however, ANN showed lower error values than RSM, confirming the robustness of
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the employed neural network algorithm over RSM. To the best of the authors’ knowledge, this
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14 research serves as the first simulation study in applying the predictive models in CBD refinement
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17 from industrial hemp. With the WFMD system being a large-scale processing equipment, this
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pioneering research documents a robust process design that yields CBD with a high
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24 concentration and high recovery rate.
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28 Compliance with current federal and state legislation: The feedstock used in this study contained
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31 less than 0.2wt.% THC content (based on dry biomass weight), which is below the maximum
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35 permitted level (MPL) designated by the current federal 46 and state 47 legislations. All the
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38 pertinent stages related to plant growth and cannabinoid processing have been under the
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41 surveillance of the industrial hemp committee of NC A&T State University.
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45 Supporting information: Additional information and characterization data, including the NMR
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49 spectra, extraction efficiency chart for different solvents, large-scale extraction setup, and HPLC
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52 chromatograms, are available free of charge via the internet at http://pubs.acs.org.
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56 41
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4 Acknowledgment: This work was supported by the United States Department of Agriculture –
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7 National Institute of Food and Agriculture (USDA–NIFA Evans Allen NC.X332-5-21-130-1
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Accession no. 1023321). The authors also would like to thank the Joint School of Nanoscience
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14 and Nanoengineering (JSNN) for supporting the research.
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18 Conflicts of Interest: The authors declare no conflict of interest.
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Refining Cannabidiol Using Wiped Film Molecular Distillationie 2022 002906.R2 Proof Hi

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Reﬁning Cannabidiol Using Wiped-Film Molecular Distillation:

Article in Industrial & Engineering Chemistry Research · May 2022

Mehrab Valizadehderakhshan Masoud Kazem-Rostami

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Arnab Bhowmik Lijun Wang

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Refining cannabidiol (CBD) using wiped-film molecular

Journal: Industrial & Engineering Chemistry Research

Manuscript Type: Article

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