Miniaturized PVC Sensors for Determination of Fe2+, Mn2+ and Zn2+ in Buffalo-Cows’ Cervical Mucus Samples
Three polyvinyl chloride membrane sensors were developed for the electrochemical evaluation of ferrous, manganese and zinc ions. The sensors were used for assaying metal ions in cervical mucus (CM) of Egyptian river buffalo-cows (Bubalus bubalis) as their levels vary dependent on cyclical hormone variation during different phases of estrus cycle. The presented sensors are based on using ionophores, β-cyclodextrin (β-CD), hydroxypropyl β-cyclodextrin (HP-β-CD) and sulfocalix-4-arene (SCAL) for sensors 1, 2 and 3 for Fe2+, Mn2+ and Zn2+, respectively. Dioctyl phthalate (DOP) was used as the plasticizer in a polymeric matrix of polyvinylchloride (PVC). For increasing the selectivity and sensitivity of the sensors, each sensor was enriched with a suitable complexing agent, which enhanced the sensor’s response. For sensor 1, β-CD was mixed with bathophenanthroline; for sensor 2, porphyrin was incorporated with HP-β-CD; while for sensor 3, oxine was the used complexing agent with SCAL. Linear responses of 10-7-10-2 M with cationic slopes of 53.46, 45.01 and 50.96 over pH range 4-8 were obtained using coated graphite sensors for ferrous, manganese and zinc ionic solutions, respectively. The three sensors were validated, according to the IUPAC guidelines. The obtained results by the presented potentiometric procedures were statistically analyzed and compared with those obtained by atomic absorption spectrophotometric method (AAS). No significant differences for either accuracy or precision were observed between the two techniques. Successful application for the determination of the three studied cations in CM, for the purpose to determine the proper time for artificial insemination (AI) was achieved. The results were compared with those obtained upon analyzing the samples by AAS. Proper detection of estrus and correct time of AI was necessary to maximize the production of buffaloes. In this experiment, 30 multi-parous buffalo-cows were in second to third lactation and weighting 415-530 kg, and were synchronized with OVSynch protocol. Samples were taken in three times around ovulation, on day 8 of OVSynch protocol, on day 9 (20 h before AI) and on day 10 (1 h before AI). Beside analysis of trace elements (Fe2+, Mn2+ and Zn2+) in CM using the three sensors, the samples were analyzed for the three cations and also Cu2+ by AAS in the CM samples and blood samples. The results obtained were correlated with hormonal analysis of serum samples and ultrasonography for the purpose of determining of the optimum time of AI. The results showed significant differences and powerful correlation with Zn2+ composition of CM during heat phase and the ovulation time, indicating that the parameter could be used as a tool to decide optimal time of AI in buffalo-cows.
 M.Guardia and S.Garrigues, Hand Book of Mineral Elements in Food, Wiley-Blackwall, UK, 2015.
 A.Bratovčić, A.Odobašić, S.Ćatić, Agriturae Consectus Scientificus 7(2) (2015) doi:10.175508.
 L.Fritea, Novel cyclodextrin modified electrodes for pharmaceutical and biomedical applications, Materials, Universit´e Grenoble Alpes, 2015. Submitted on 15 Oct 2015.
 S.S. Hassan, M.M. Amer, S.A. Abd El-Fattah, A.M. El-Kosasy, Talanta 46 (1998) 1395-1403.
 A.M. EL-Kosasy, M.Y. Salem, M.G. El-Bardicy, M.K. Abdelrahman, Chem. Pharm. Bull. 56 (2008) 753-757.
 T.F. Sousa, C.G. Amorim, M.C.B.S.M. Montenegro, A.N. Araújo, Sensors and Actuators B 176 (2013) 660-666.
 A.M. El-Kosasy, M. Nebsen, M.K. Abd El-Rahman, M.Y. Salem, M.G. El-Bardicy, Talanta 85 (2011) 913-918.
 M.A. El-Sayed, Sensors and Actuators B 190 (2014) 101-110.
 M. Nebsen, G.M. El-Sayed, M. Abdel Kawy, S.Z. El-Khateeb, Anal. Bioanal. Electrochem. 5(3) (2013) 368-380.
 J. Roelofs, F. Gatius, R. Hunter, F.V. Eedenburg, C. Hanzen, Theriogenology 74(3) (2010) 327-344.
 T. Rao, N. Khumar, P. Khumar, S. Chaurasia, N. Patel, Vet. World 6(6) (2013) 363-369.
 M. Jainudeen and E. Hafez, Cattle and buffalo. In: Hafez ESE, editor. Reproduction in farm animals. 6th ed., Philadelphia: Lea and Febiger. (1993) 315-329.
 M. Jainudeen, Reproduction problems of buffaloes in the world. Proceedings II World Buffalo Congress, New Delhi III: (1988) 189-196.
 R. Scipioni and R. Foote, Journal of Dairy Science 82 (1999) 1742-1745.
 S. Layek, A. Kumaresan, K. Behera, S. Chand, Science Cattle 152(2) (2013) 273-281.
 R. Predojevici, T. Petrujki, M. Predojevi, LucrariStinifice Medicine Veterina, Vol. XI, Timisoara (2007) 91-94.
 M.M. Eltohamy, A.D. Zakaria, N.A. Taha, Animal Reproduction Science 22(3) (1990) 203-211.
 A.Peters, and P. Ball, Reproduction in cattle, Blackwell Science, Oxford, UK (1995) 170-182.
 L. Modi, B. Suthar, C. Chaudhari, N. Chaudhari, H. Nakhashi, M. Falguni, Vet World 6(3) (2012) 143-146.
 J. Das, P. Dutta, C. Deka, R. Biswas, B. Sarmah, A. Dhali, Livestock Research for rural development, 21(5) (2009).
 S. Anas, A. Narsat, M. Gamal, Frontiers in Bioscience E5 (2013) 798-808.
 M.K. Shepard, and Y.D. Senturia, Fertility and Sterility 28 (1977) 541-548.
 E.Norkus,J. Incl Phenom Macrocycl Chem 65(2009) 237-248
 IUPAC, Analytical Chemistry Division, Commission on Analytical Nomenclature, Pure Appl. Chem. 72 (2000) 1851.
 A.Manda, B. Gereksinim, S. Kalitesi, E. Uretimi, Kocatepe Vet J. 3(2) (2010) 55-64.
 J.S. Stevenson, Y. Kobayashi, K.E. Thompson, J. Dairy Sci. 82 (1999) 506-515.
 F. Fernandez and H. Kohn, ClinChem. 13 (1991) 1096-101.
 R. Ungaro, A. Arduini, A. Casnati, A. Pochini, F. Ugozzoli. Pure and App. Chem. 68(6) (1996) 1213-1218.
 R. Ludwig and N. Thi K. D. Sensors 2 (2002) 397-416
 L. Chen, J. Zhang, W. Zhao, X. He, Y. Liu, J. Electroanal. Chem. 589 (2006) 106-110.
 M. Zareh, B. Malinowska, J. AOAC Int. 90(1) (2007) 147-152.
 F. Mizani, M. Majdi, S. Taghvaei, Anal. Bioanal. Electrochem. 4(5) (2012) 529-543.
 M. Biesaga, K. P. Ska, M. Trojanowicz, Talanta 51 (2000) 209-224.
 Z. Shi, C. Fu, Talanta 44(4) (1997) 593-604.
 V.K. Gupta, D.K. Chauhan, V.K. Saini, S. Agarwal, M.M. Antonijevicand, H. Lang, Sensors 3 (2003) 223-235.
 S. Chandra, D. R. Singh, Journal of Saudi Chemical Society 14 (2010) 55-60.
 E.M. Rakhman’ko, V.V. Egorov, M.Ya. Tarazevich, A.D. Rubinova, Journal of Analytical Chemistry 58(7) (2003) 691-697.
 Q. Fu, S. Qian, Na Li, Q. Xia, Y. Ji, Int. J. Electrochem. Sci. 7 (2012) 6799-6806.
 S. Amemiya, P. Bühlmann, Y. Umezawa, Anal. Chem. 70 (1998) 445.
 P.Bühlmann, Y. Umezawa, Electroanalysis 11 (1999) 687.
 G. Snedecor, and W. Cochran, Statistical methods 8th edition. Iowa State University. Press, Ames, Iowa, USA (1987).