Animal trials were conducted to determine the bioavailability of the TurmiZn complex in rats and chickens. Rats are a more suitable mammalian model than mice to perform kinetic studies that require blood withdrawal for longer durations. Chickens are suitable models for bioavailability studies for researchers interested in avian production. Doses were selected based on the ability to readily monitor TurmiZn metabolites analytically in plasma and were not based on therapeutic levels.
For the avian bioavailability trials, chickens (Rhode Island Red, 3 chickens/group) that were each 2 years old and weighing 5 pounds were selected, and 1 mL of blood was drawn from the wing vein for baseline measurements. The blood was allowed to clot and the plasma was removed by centrifugation for subsequent baseline analysis. The chickens were allowed to rest overnight. One gram of TurmiZn, one gram of curcumin, and one gram of THC were suspended in 5 mL of water and placed into respective syringes. The contents of each of the respective syringes were then administered orally ensuring that the contents were consumed. Blood was drawn at 2, 4, and 24 h from each bird and allowed to clot. The recovered serum was mixed with equal volumes of acetone via vortexing for 30 s and then centrifuged to remove proteins [34 (link),35 (link)] in the pellet while recovering curcuminoids, THC, or elements of TurmiZn in the supernatant. The supernatants from each time point were analyzed by HPLC methodology as described for the presence of curcuminoids, THC, or TurmiZn at 425 nm.
Bioavailability study in rats was approved by IAEC (Institutional Animal Ethics Committee) of the Institute of Pharmacy, Nirma University (Ahmedabad, India) as per the CPCSEA (Committee for Control and Supervision of Experiments on Animals) guidelines, Department of Animal Husbandry and Dairying (DAHD), Ministry of Fisheries, Animal Husbandry and Dairying (MoFAH&D), Govt. of India. The protocol number was IP/PCOL/FAC/29/2021/32. The bioavailability was evaluated in adult healthy male Wistar rats with two different studies: oral administration and intravenous administration of TurmiZn complex as a single dose. For orally administered trials in mammals, baseline blood samples were taken and three animals were administered TurmiZn 5 g by mouth. Blood samples were then withdrawn at 15-minute intervals up to 60 min and then hourly for up to 4 h and allowed to clot. The recovered serum was mixed with equal volumes of acetone via vertexing for 30 s and then centrifuged to remove proteins in the pellet while recovering elements of TurmiZn in the supernatant. The samples were analyzed by HPLC and liquid chromatography-mass spectrometry/mass spectrometry (LC/MS/MS). In another study, intravenous (IV) injection of TurmiZn in a single healthy adult Wistar rat was used to obtain metabolic profiles of TurmiZn over time. This was based on a protocol [36 (link)] for the pharmacokinetics and tissue distribution of curcumin in mice. TurmiZn was injected (100 mg/Kg, IV) and blood was collected at 2, 5, 15, 30, 45, and 60 min and allowed to clot and extracted as previously described in acetone for ultra-fast liquid chromatography (UFLC), LC-MS/MS analysis.
For the avian bioavailability trials, chickens (Rhode Island Red, 3 chickens/group) that were each 2 years old and weighing 5 pounds were selected, and 1 mL of blood was drawn from the wing vein for baseline measurements. The blood was allowed to clot and the plasma was removed by centrifugation for subsequent baseline analysis. The chickens were allowed to rest overnight. One gram of TurmiZn, one gram of curcumin, and one gram of THC were suspended in 5 mL of water and placed into respective syringes. The contents of each of the respective syringes were then administered orally ensuring that the contents were consumed. Blood was drawn at 2, 4, and 24 h from each bird and allowed to clot. The recovered serum was mixed with equal volumes of acetone via vortexing for 30 s and then centrifuged to remove proteins [34 (link),35 (link)] in the pellet while recovering curcuminoids, THC, or elements of TurmiZn in the supernatant. The supernatants from each time point were analyzed by HPLC methodology as described for the presence of curcuminoids, THC, or TurmiZn at 425 nm.
Bioavailability study in rats was approved by IAEC (Institutional Animal Ethics Committee) of the Institute of Pharmacy, Nirma University (Ahmedabad, India) as per the CPCSEA (Committee for Control and Supervision of Experiments on Animals) guidelines, Department of Animal Husbandry and Dairying (DAHD), Ministry of Fisheries, Animal Husbandry and Dairying (MoFAH&D), Govt. of India. The protocol number was IP/PCOL/FAC/29/2021/32. The bioavailability was evaluated in adult healthy male Wistar rats with two different studies: oral administration and intravenous administration of TurmiZn complex as a single dose. For orally administered trials in mammals, baseline blood samples were taken and three animals were administered TurmiZn 5 g by mouth. Blood samples were then withdrawn at 15-minute intervals up to 60 min and then hourly for up to 4 h and allowed to clot. The recovered serum was mixed with equal volumes of acetone via vertexing for 30 s and then centrifuged to remove proteins in the pellet while recovering elements of TurmiZn in the supernatant. The samples were analyzed by HPLC and liquid chromatography-mass spectrometry/mass spectrometry (LC/MS/MS). In another study, intravenous (IV) injection of TurmiZn in a single healthy adult Wistar rat was used to obtain metabolic profiles of TurmiZn over time. This was based on a protocol [36 (link)] for the pharmacokinetics and tissue distribution of curcumin in mice. TurmiZn was injected (100 mg/Kg, IV) and blood was collected at 2, 5, 15, 30, 45, and 60 min and allowed to clot and extracted as previously described in acetone for ultra-fast liquid chromatography (UFLC), LC-MS/MS analysis.
Free full text: Click here
