In Situ Single-Pass Perfused Rabbit Intestinal Study Sample Analysis of Ractopamine by UPLC

Fatima. Namzi

National College of Pharmacy, KMCT Group of Institutions, Manassery, Kozhikode-673602, Kerala, India.

Kathirvel. Singaram

National College of Pharmacy, KMCT Group of Institutions, Manassery, Kozhikode-673602, Kerala, India.

Raja Rajeswari. Katta *

Department of Pharmaceutical Analysis, Sri Sivani College of Pharmacy, Srikakulam-532 402, Andhra Pradesh, India.

*Author to whom correspondence should be addressed.


Abstract

Researchers have struggled in recent years to anticipate how oral drugs would be absorbed by people, and numerous permeability study approaches have been created for a variety of uses, including biowaiver procedures. The Single-Pass Intestinal Perfusion (SPIP) technique, used on rats, can produce permeability results that are most similar to in vivo circumstances. Since SPIP in rats is the closest approximation to in vivo settings, calculations for absorption prediction in humans and SPIP in rats may show the transport pathways and/or pre-systemic metabolism implicated on permeation processes of pharmaceuticals. In order to increase feed efficiency and promote growth in cattle, ractopamine hydrochloride, a commercial beta-adrenergic agonist, is frequently used as a dietary supplement. Currently, muscle and liver are the only regulatory target tissues (as authorised in the New Animal Drug Application with the Food and Drug Administration) for ractopamine residue testing. The study's goal is to create and validate a UPLC assay method to assess whether or not perfused ractopamine is detectable and quantifiable in cow small intestine. Acquity UPLC BEH C18 (150 mm x 2.1 mm, 1.7 µm) reversed phase column is used for the separation. Acetonitrile and 0.01M ammonium formate are used as the mobile phase in gradient mode with a flow rate of 0.5 mL/min. The guiding principles are satisfied by the precision, accuracy, extraction recovery, matrix effect, and stability. The created approach was successfully applied to a perfusion research in the small intestine of a rabbit, and the samples were analysed by Empower software, in order to verify the sensitivity and selectivity of the method in a real-time context. Ractopamine has a Peff value of 0.022 0.003 x 10-4, indicating that the jejunum region has minimal permeability for it.

Keywords: UPLC, Ractopamine, Perfusion, Small Intestine, Validation


How to Cite

Namzi , Fatima., Kathirvel. Singaram, and Raja Rajeswari. Katta. 2023. “In Situ Single-Pass Perfused Rabbit Intestinal Study Sample Analysis of Ractopamine by UPLC”. Asian Journal of Research in Nephrology 6 (1):101-13. https://journalajrn.com/index.php/AJRN/article/view/68.

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References

Colbert WE, Williams PD, Williams GD. Beta-adrenoceptor profile of ractopamine HCl in isolated smooth and cardiac muscle tissues of rat and guinea-pig. The Journal of Pharmacy and Pharmacology. 1991;43(12):844–847. DOI:10.1111/j.2042-7158.1991.tb03192.

Liu X, Grandy DK, Janowsky A. Ractopamine, a livestock feed additive, is a full agonist at trace amine-associated receptor 1. The Journal of Pharmacology and Experimental Therapeutics. 2014;350(1):124–129. DOI:10.1124/ jpet.114.213116.PMC 4170122.

Mersmann HJ. Overview of the effects of beta-adrenergic receptor agonists on animal growth including mechanisms of action. Journal of Animal Science. 1998;76:160-172. DOI: 10.1016/j.aca.2004.09.053.

Wang JP, Zhang SX, Shen JZ. Technical note: A monoclonal antibody-based immunoassay for determination of ractopamine in swine feeds,” Journal of Animal Science. 2006;84:1248–1251.

Guanglong D, Li D, Qin J, Zhu J, Wang B, Geng Q, Guo M, Punyapitak D, Cao Y. Development and validation of a high-performance liquid chromatography method for determination of ractopamine residue in pork samples by solid phase extraction and pre-column derivatization. Meat Science. 2015;106:55-60.

Sakai T, et al. Determination method for ractopamine in swine and cattle tissues using LC/MS". Shokuhin Eiseigaku Zasshi. 2007;48(5):144–147. DOI:10.3358/shokueishi.48.144

Antignac JP, Marchand P, Le Bizec B, Andre F. Identification of ractopamine residues in tissue and urine samples at ultra-trace level using liquid chromatography-positive electrospray tandem mass spectrometry. Journal of Chromatography B. 2002;774:59–66.

Haley ED, Crystal-Dawn B, Patrick B, Ifigenia G, Thomas JB, John S, Keith B, Jessica P. Quantification of ractopamine residues on and in beef digestive tract tissues, Journal of Animal Science. 2019;97(10):4193–4198.

Nie JT, Hong ZT, Lian J, Deng XL. Rapid determination of ractopamine and clenbuterol in animal urine with HPLC-MS/MS. Chin. J. Vet. Drug. 2008;42(8):17–20.

Ying YF, Pi XE, Wu PG, Chen HH, Zhu CY, Lu CB. Simultaneous determination of ractopamine and clenbuterol residues in Animal Urine with Gas Chromatography-Mass Spectrometry. J. Chin. Mass Spectr. Soc. 2006;27(2):74–78.

Food and drug administration. Guidance for Industry: Bioanalytical Method Validation Center for Drug Evaluation and Research. Silver Springs, MD: FDA; 2001.

Amruta SK, Nayana VP, Puja SG, Sushila DC. Bioanalytical method validation. Asian J. Pharm. Ana. 2015;5(4):219-225.

ICH guideline M10 on bioanalytical method validation and study sample analysis EMA/CHMP/ICH/172948/2019.

VICH guideline 49 studies to evaluate the metabolism and residue kinetics of veterinary drugs in food-producing animals, U.S. food and drug administration, center for veterinary medicine; 2011. Available:http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/GuidanceforIndustry/UCM207942.pdf

Pereira PF, Wojnowski W, Tobiszewski M. AGREE - Analytical GREEnness Metric Approach and Software. Anal Chem. 2020;92(14):10076–82.

Gałuszka A, Migaszewski ZM, Konieczka P, Namieśnik J. Analytical Eco-Scale for assessing the greenness of analytical procedures. TrAC - Trends Anal Chem. 2012;37:61–72.