February Survey on the Concentrations of Hormones (Estradiol-17b, Progesterone, Testosterone) in Bovine Muscles

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1 February b, , Survey on the Concentrations of Hormones (,, ) in Bovine Muscles Tomoyuki M>N6O6@> and Tsuneo H6H=>BDID (Tokyo Metropolitan Institute of Public Health: , Hyakunin-cho, Shinjuku-ku, Tokyo , Japan; Corresponding author) Natural sex hormones, estradiol-17b (EST), progesterone (PRO) and testosterone (TES), in sixty domestic bovine muscles and forty bovine muscles imported from USA and Australia were determined by radioimmunoassay. The EST, PRO and TES levels (mean standard deviation) in domestic samples (n 60) was ppt, ppb and ppt on a whole basis, respectively. The hormone levels in muscles was presumed to reflect sex, estrous cycle, and so on. The hormone levels in imported bovine muscles (n 40) were ppt (EST), ppb (PRO) and ppt (TES), respectively. (Received August 28, 2003) Key words: bovine muscle; -17b estradiol-17b; progesterone; testosterone; hormone level; RIA radioimmunoassay -17b, -17b, 1 1) ) (4 : 1) Sep-Pak Vac C 18 Bond Elut DEA 1) 10.0 g 10 ml, (1 : 2) 80 ml

2 50 3) 1ppt, 0.04 ppb, 10 ppt 4) 30 1) ppb 4 /60 (6.6 ) 10 ppt 32 /60 (53.3 ) -17b 1ppt 44 / 60 (73.3 ) Table 1-17b 0.87 ppt, 1.73 ppt, 2.62 ppb, 4.34 ppb, 16.6 ppt, 59.5 ppt 2 Vol. 45, No. 1 2) Table 2-17b 1.51 ppt, 0.80 ppt 6.00 ppb, 0.37 ppb 22.7 ppt, 39.0 ppt -17b, b (12.8 ppt) (952 ppt) r 0.67 r 0.49 r ) Table 3-17b 0.87 ppt 2.68 ppt, 1.58 ppt, 7.40 ppb, 4.31 ppb, 3.89 ppb 24.9 ppt, 15.8 ppt, 37.0 ppt -17b 3 Table 1. Hormone Concentrations in Muscles of Domestic Cattle Species Number of samples Holstein-Fresian ( ) (13.9) (0.0418) (1075.4) Japanese Black Cattle (Wagyu) ( ) (112.8) (0.0436) (10952) 1) Detection limits: 1 ppt: 0.04 ppb: 10 ppt 2) Upper column indicates mean S.D., Lower column indicates concentration range Table 2. Hormone Concentrations in Muscles of Domestic Cattle by Sex Sex Number of samples Female [Heifer] ( ) (112.8) (0.0418) (1049.4) Male [Steer] ( ) (15.8) ( ) (10952)

3 February Table 3. Hormone Concentrations in Muscles of Heifers in Estrous Cycle Estrous cycle Number of samples 2 2) 4) 20 Table 4 No., -17b 3.11 ppt, 3.56 ppt, 0.55 ppb, 0.50 ppb, 8.51 ppt, 9.05 ppt Luteal phase (12.6) ( ) (1075.4) Follicular phase (112.8) (1.7 18) ( ) Uncertain (15.2) ( ) (1015.2) 2 5) Table 5-17b ppt, ppt, 3.19 ppb, 0.52 ppb 30.9 ppt, 8.8 ppt JECFA FAO/WHO ) Arnold 1-17b (Steer) pg/g 2.1 pg/g Heifer, pg/g, 5.80 pg/g RCBA (Recombinant Cell yeast Bioassay) 4) 6) 6) ADI 52 JECFA 3) 32 JECFA 7) ADI Acceptable Daily Intake: (Table 6) 0 50 ng/kg/day, 0 30 mg/kg/day, 0 2 mg/kg/day Table 4. Hormone Concentrations in Muscles of Imported Cattle Country Number of samples Australia ( ) (19.8) ( ) (1032.4) USA ( ) (110.0) ( ) (1078.4) 1 Arnold, D., Hormones, revised working paper on residues ( ); JECFA 1999.

4 52 Vol. 45, No. 1 Table 5. Hormone Concentrations in Muscles of Imported and Domestic Cattle Sample Number of samples Domestic ( ) (112.8) (0.0436) (10952) Imported ( ) (110.0) ( ) (1078.4) Table 6. ADI of Hormones Established at the 52nd Meeting of JECFA ( ) Compound No-Observed-E#ect Level (NOEL) 0.3 mg/day (5 mg/kg bw) 200 mg/day (3.3 mg/kg bw) 100 mg/day (1.7 mg/kg bw) Safty Factor (SF) Acceptable Daily Intake (ADI) ng/kg mg/kg 1, mg/kg MRLs (Maximum Residue Limits): not specified in cattle muscle, liver, kidney, or fat MRLs Maximum Residue Limits: 50 kg 1 2,500 ng/day, 1,500 mg/day, 100 mg/day -17b 12.8 ppt (ng/kg) kg -17b -17a ) 9) 11) EU 11) 11 12) 10 13) 1) Hashimoto, T., Jimbo, K., Miyazaki, T., Simple preparation for determination of steroid hormones in beef by radioimmunoassay. Tokyo Eiken Nenpo, 53, (2002). 2) Domeki, I., Rinsho Jui (J. Clin. Vet. Med.), 4, (1986). 3) Joint FAO/WHO expert committee on Food additives, WHO technical report series 893, Evaluation of certain veterinary drug residues in food. Fifty-second report, 2000, p ) Klein, K. O., Baron, J., Colli, M. J., McDonnell, D. P., Cutler, Jr., G. B., Estrogen levels in childhood determined by an ultra-sensitive recombinant cell bioassay. J. Clin. Invest., 94, 2,475 2,480 (1994). 5) Coldham, N. G., Evaluation of a recombinant yeast cell estrogen screening assay. Environ. Health Perspect., 105, (1997). 6) Burdge, G. C., Determination of estrogen concentration in bovine plasma by a recombinant estrogen receptorreporter gene yeast bioassay. Analyst, 123, 2,585 2,588 (1998). 7) Joint FAO/WHO expert committee on Food additives, WHO technical report series 763, Evaluation of certain veterinary drug residues in food. Thirty-second report, 1988, p ) Hartmann, S., Lacorn, M., Steinhart, H., Natural occurrence of steroid hormones in food. Food Chemistry, 62, 7 20 (1998). 9) Maume, D., Deceuninck, Y., Pouponneau, P., Paris, A., Le Bizec, B., Andre, F., Assessment of estradiol and its metabolites in meat, APMIS, 109, (2001). 10) Andersson, A. M., Skakkebaek, N. E., Exposure to exogenous estrogenes in food: possible impact on human development and health. Eur. J. Endocrinol., 140, (1999). 11) Stephany, R. W., Hormone in meat: di#erent ap-

5 February proaches in the EU and in the USA. APMIS Suppl., 103, S (2001). 12) 11 ( ). 13) 10 ( ).