Excretion and residue depletion of cannabinoids in beef cattle fed hempseed cake for 111 days.

Thirty-two crossbred heifers were fed either a control diet or 20% (dry matter basis) hempseed cake in a complete ration for 111 days; of the cattle fed hempseed cake, four each were harvested with 0, 1, 4, and 8-day withdrawal periods. Urine and plasma were collected during the feeding and withdrawal periods and liver, kidney, skeletal muscle, and adipose tissue were collected at harvest. Total cannabinoid (n = 10) concentration of hempseed cake averaged 11.3 ± 11.7 mg kg-1 across the feeding period with total cannabidiol and tetrahydrocannabinol (CBD/THC) concentrations of 1.3 ± 0.8 mg kg-1. Neutral cannabinoids (cannabinol [CBN], CBD/THC, and cannabidivarin [CBDV]) were not detected in plasma or urine, but CBD/THC was measured in adipose tissue at all withdrawal periods (6.3 ± 2.1 to 10.1 ± 2.5 ng g-1). In contrast, cannabinoid acids (cannabinolic acid [CBNA], cannabidiolic acid [CBDA]/tetrahydrocannabinolic acid [THCA], cannabichromenic acid [CBCA], and cannabidivarinic acid [CBDVA]) were sporadically detected (<15 ng mL-1) in plasma and urine of cattle fed hempseed cake. Cannabinoid acids were depleted from liver by withdrawal day 4, but could still be measured (<1 ng g-1) in kidney of some animals harvested on withdrawal day 8. Assessment of human exposures to CBD/THC residues through the consumption of beef fat from animals fed hempseed cake suggests that the probability of consuming the equivalent of an acute reference dose (ARfD) is remote, even with the use of a conservative reference dose ARfD (1 µg kg-1 body weight).

Rapid quantification of cannabinoids in beef tissues and bodily fluids using direct-delivery electrospray ionization mass spectrometry.

Hempseed cake is a byproduct of hempseed oil extraction and is potentially a useful source of protein and fiber for use in ruminant diets. However, data are lacking on the appearance and/or clearance of cannabinoids in tissues of animals fed hempseed cake. To this end, a rapid method for quantifying cannabinol (CBN), cannabidiol (CBD), cannabinolic acid (CBNA), cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), cannabichromenic acid (CBCA), cannabidivarin (CBDV), cannabidivarinic acid (CBDVA), tetrahydrocannabinol (THC) and tetrahydrocannabinolic acid (THCA) in cattle tissues, plasma, and urine was developed using rapid screen electrospray ionization mass spectrometry (RS-ESI-MS). Regression coefficients of matrix-matched standard curves ranged from 0.9946 to >0.9999 and analyte recoveries averaged from 90.2 ± 15.5 to 108.7 ± 18.7% across all compounds. Limits of detection and quantification ranged from 0.05 to 2.79 ng · mL-1 and 0.17 to 9.30 ng · mL-1, respectively, while the inter-day relative standard deviation ranged from 5.1 to 15.1%. Rapid screening electrospray ionization mass spectrometry (RS-ESI-MS) returned no false positives for any cannabinoid in plasma, urine, and tissue (liver, skeletal muscle) samples from 6 non-dosed control animals (n = 90 samples; of which 72 samples were plasma or urine and 18 samples were tissues). Across-animal cannabinoid concentrations measured in 32 plasma samples of cattle dosed with ground hemp were quantified by RS-ESI-MS; analytical results correlated well (r2 = 0.963) with independent LC-MS/MS analysis of the same samples.

Influence of hempseed cake inclusion on growth performance, carcass characteristics, feeding behavior, and blood parameters in finishing heifers.

As the hemp industry continues to develop in the United States, there is an increasing interest in feeding byproducts of industrial hemp production to livestock. A completely randomized design experiment using crossbred finishing heifers (initial body weight [BW] ±â€…SE = 494 ±â€…10 kg) was conducted to determine the effects of feeding hempseed cake in a corn-based finishing diet (10% forage) formulated to meet or exceed ruminally degradable and metabolizable protein requirements on growth performance, carcass characteristics, feeding behavior, and plasma parameters. Dietary treatments were the inclusion of 20% (dry matter [DM] basis) of dried corn distillers grains plus solubles (DDGS, n = 16) or hempseed cake (HEMP, n = 15). Cattle were housed in two pens, had ad libitum access to feed and water, and individual intakes and feeding behavior were monitored using the Insentec feeding system. Cattle were fed treatment diets for 111 d, and every 14 d BW were measured and blood samples were collected. Blood plasma was analyzed for glucose, urea nitrogen, and individual amino acids, and results were analyzed using repeated-measures analysis in SAS. Final BW, average daily gain, gain:feed, and hot carcass weight decreased (P ≤ 0.05) by 2.3%, 7.7%, 7.7%, and 2.6%, respectively, in heifers fed the HEMP diet than in heifers fed the DDGS diet. Net energy for maintenance and gain (Mcal/kg of feed, DM basis), estimated based on heifer intake and performance, were greater (P = 0.02) for the DDGS diet than for the HEMP diet. All other performance and carcass characteristics were not different (P ≥ 0.20) between treatments. Heifers fed the HEMP diet had greater (P < 0.05) plasma urea nitrogen concentration in samples from each collection day compared with heifers fed the DDGS diet, although there was a treatment-by-day interaction (P < 0.01) because of variability in the magnitude of treatment differences over time. Plasma glucose concentration was not influenced (P = 0.17) by dietary treatment. Plasma concentrations of total amino acids, nonessential amino acids, and essential amino acids were not different between treatments (P ≥ 0.09), although there were several interactions between treatment and day (P ≤ 0.04) for individual amino acids. These data suggest that hempseed cake has a lower net energy for maintenance and gain relative to DDGS when adequate metabolizable protein is supplied, while still providing adequate nutrition to support the acceptable performance of finishing cattle.

This experiment evaluated the effects of dietary inclusion of hempseed cake in comparison with dried corn distillers grains plus solubles (DDGS) on growth performance, feeding behavior, carcass characteristics, and blood parameters in finishing heifers. Angus-crossbred heifers were assigned randomly to one of two treatments (20% hempseed cake [HEMP] or 20% DDGS; dry matter basis) and were fed for 111 d until slaughter. Heifers receiving the DDGS treatment had greater final body weights, average daily gain, gain efficiency, dietary concentration of net energy for maintenance and gain, and carcass weight than heifers fed the HEMP treatment. All other carcass characteristics, as well as feeding behavior, were not influenced by treatment. Plasma urea nitrogen was greater in heifers fed the HEMP diet compared with the DDGS diet, while glucose was not influenced by treatment. Several plasma amino acid concentrations were influenced by treatment. Although the inclusion of hempseed cake decreased growth performance, it could be a viable alternative feed source for cattle.

Structure-activity relationships of the prototypical TRPM8 agonist icilin.

A series of structural analogues of the TRPM8 agonist icilin was prepared. The compounds were examined for their ability to exert agonist or antagonist effects in HEK-293 cells expressing the TRPM8 receptor. Most structural modifications of the icilin structure largely met with diminished TRPM8 agonist activity. Cinnamamide 'open-chain' analogs of icilin, however, demonstrated significant antagonistic actions at the TRPM8 receptor. Optimal potency (IC50=73 nM) was observed in the 3-iodo derivative 18l.