Residues of anabolic agents in Red Meat and Meat Products Using HPLC and ELISA Isam T. Kadim Department of Animal and Veterinary Sciences College of Agricultural and Marine Sciences Sultan Qaboos University 2nd International Conference on Food Safety and Regulatory Measures, June 06-08, 2016, London, UK

♦ Increasing global distribution of food ♦ Countries more dependant on each other for their food supply ♦ Mass production (increase population) Consumers demand: ♦ a nutritious product ♦ with minimal microbial contamination ♦ no chemical residues above acceptable limits ♦ no parasites ♦ beneficial to human health

Chemical Contaminants ♦ Intentional Food Additives ♦ Use of materials which enhance the acceptability of the products and/or aid in the development of the product ♦ Unintentional Food Additives ♦ Residues ♦ Includes both food processing component residues as well as PESTICIDES, ANTIBIOTIC OR HORMONE IMPLANT residues ♦ Natural toxicants ♦ Product tampering ♦ Nutritional components

♦ Chemical contaminants considered number one hazard in human food ♦ 400 million tons of chemicals produced annually worldwide ♦ Meat and meat products may contain harmful residues ♦ Food additives are a major cause of cancer in the world

What are contaminants? ♦ Any biological, chemical property that may cause food to be unsafe for human consumption ► Microbial ► Chemical ► Biological Maximum Residue Limits (MRLs)

Contaminants from Farm to Consumers Pesticides Agriculture products Processing Transport Storage Feed Environment Hormones Antibiotic Medication Display Aflatoxin Disinfectants Heavy metals Melamine Microbial Slaughtering

Chemical Contamination ♦ Veterinary drugs (600 substances) ♦ Pesticides (600 substances) ♦ Heavy metals ♦ Melamine ♦ Alfatoxin

Biological Contaminants “Biological” means “living” ♦ Bacteria ♦ Viruses ♦ Parasites ♦ Fungi –Moulds –Yeasts

Risk of Contamination of Animal Products ♦ ♦ ♦ ♦ ♦ ♦ Contamination of pasture, drinking water and air ♦ Growth promoters and veterinary medicines ♦ Deposition of harmful substances in meat, fat, liver, kidney, brain and spleen ♦ Improper meat processing can cause contamination ♦ Contamination of meat and meat products during the processing and food cycle

♦ Although cost benefits using hormones in meat production, no compromise to human health ♦ Hormones naturally occurring substances ♦ Hormone residues should be below the MRLs ♦ MRLs safety measures based on Accepted Daily Intakes (ADIs) ♦ Estrogen, progesterone and testosterone hormoned naturally present in red meat ♦ These hormones also present in humans

11 Growth-Promoting Implants ♦ Used in animals to: ♦ increase rate of growth & weight gain ♦ increase feed efficiency ♦ increase muscle growth and influence carcass traits ♦ increase meat mass and reduce fat ♦ improve nutrient absorption ♦ improve meat quality

12 ♦ Contain either natural or synthetic hormones ♦ natural hormones are gender-based hormones ♦ estradiol (esterogen) ♦ progesterone ♦ testosterone ♦ synthetic hormones are based on natural hormones but modified in a lab ♦ zeranol ♦ melengestrol acetate ♦ trenbolone acetate ♦ animal must not have received the compound or drug immediately preceding harvest ♦ allow time for the hormone to pass via animal’s system Growth-Promoting Implants

♦ I mportant food commodity which provides nutrients essential for human health ♦ S upply high biological value protein, iron, zinc, and vitamin B 12 contents ♦ Oman is an open market for a variety of meat types from different countries ♦ Meat and meat products are integral parts of the Omani daily diet Why Meat and Meat Products

Meat Consumption Rate (kg Capita/year) ♦ Global meat consumption (approximately 41.3 kg/year/person) ♦ Meat consumption in developing countries (31.6 kg/year) ♦ Meat consumption in developed countries (85.8 kg/year) ♦ Arab countries are rich in livestock ♦ High meat consumption ♦ Open market for a variety of food ♦ Low quality of animal products FAOSTAT, 2015

Objectives ♦ Determine levels of anabolic agents ♦ Testosterone ♦ Estrogen ♦ Trenbolone ♦ Fresh, frozen, minced, sausages, burgers & mortadella ♦ Beef ♦ Buffalo ♦ Sheep

Sample Collection 81 red meat samples Beef, Buffalo and Sheep Fresh, Frozen and Processed Minced, burger, sausages, mortadella

HPLC ELISA Two methods used to determine hormones in meat and meat products

Determination of hormones Extraction triplicates Following standard procedure ReadingELISA HPLC

RESULTS Hormone residues (ppb) in fresh beef meat representing two brands available in the local Omani market ND: Not detected

RESULTS Hormone contaminates residues (µg/L) in fresh sheep meat representing two brands available in the local Omani market

RESULTS Hormone residues (µg/L) in frozen beef, and sheep meats representing three different brands available in the local Omani market.

RESULTS Hormone residues (ppb) in frozen buffalo meat representing five brands available in the local Omani market

RESULTS Hormone pesticide residues (ppb) in beef and buffalo minced meat representing various brands available in the local Omani market

RESULTS Hormone residues (ppb) in beef sausage representing three brands available in the local Omani market ●●● The value of concentration is higher than the highest concentration of the standard dilution (2000 ppb for testosterone )

Conclusion ♦ Residues of testosterone and estrogen were above the maximum residual limits (MRLs) ♦ Anabolic residues and their metabolites in meat products may cause adverse toxic or carcinogenic effects on consumers’ health ♦ A monitoring program needs to be carried out to prohibit distribution of foodstuffs that could pose a risk to human health if consumed ♦ There should be a criteria for identification and confirmation of various contaminants in meat and meat products, for compliance with rules and regulations

Thank you