Sintesis Konjugat Bsa-Hapten Asam 4-(4,6-Diamino-1,3,5-Triazin-2- Ylamino) Butanoat Untuk Pengembangan Deteksi Selektif Melamin
DOI:
https://doi.org/10.21580/wjc.v3i1.6134Keywords:
melamin, sintesis, hapten, konjugat BSA-HaptenAbstract
Sejak isu kontaminasi melamin pada produk susu diketahui secara luas, diperlukan penelitian terbaru tentang teknik pemantauan yang cepat dan akurat untuk mendeteksi melamin dalam produk susu. Suatu metode indirect competitive enzyme-linked immunosorbent assay (icELISA) dengan spesifisitas yang tinggi dikembangkan untuk deteksi melamin dalam susu. Sintesis hapten dan pembentukan konjugat hapten-protein dilakukan sebagai immunogen untuk membentuk antibodi yang selektif terhadap melamin. Hapten telah disintesis dari asam gamma-amino butirat (GABA) dengan 2- kloro-4,6-diamino-1,3,5-triazin (CAAT). Digunakan disikloheksilkarbodiimida (DCC) dan N-hidroksisuksinimida (NHS) untuk merubah gugus karboksilat hapten menjadi ester aktif sehingga dapat dikonjugasikan ke bouvine serum albumin (BSA). Hapten asam 4-(4,6-diamino-1,3,5-triazin-2-ylamino)-butanoat berhasil disintesis dengan yield 8,16%, dan serapan maksimum UV pada panjang gelombang 233 nm. Karakterisasi hapten menunjukkan adanya serapan inframerah gugus amina sekunder pada bilangan gelombang 1654,92 cm-1 dan spektrum mass spectroscopy pada m/z 212,4 yang menunjukkan massa relatif hapten (C7H12N6O2). Konjugat hapten-BSA menunjukkan serapan UV maksimum pada panjang gelombang 215 nm. Hasil karakterisasi menunjukkan hapten dan konjugat hapten-BSA telah berhasil disintesis.
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Cao B., Hong Y., Juan S., Huafang C., Shuqun Li., Anping D. 2013. Sensitivity and Specificity Enhanced Enzyme-Linked Immunosorbent Assay by Rational Hapten Modification and Heterogeneous Antibody/Coating Antigen Combinations for the Detection of Melamine in Milk, Milk Powder and Feed Samples. Talanta 116 : 173–180
Lei, H.T., Shen, Y.D., Song, L.J., Yang, J.Y., Chevallier, O.P., Haughey, S.A., Wang, H., Sun, Y.M., Elliott, C.T. 2010. Hapten synthesis and antibody production for the development of a melamine immunoassay. Anal. Chim. Acta., vol. 665 : 84-90.
Liu, J.X., Zhong, Y.B., Liu, J. 2010. An enzyme linked immunosorbent assay for the determination of cyromazine and melamine residues in animal muscle tissues. Food Control, vol. 21 : 1482-1487.
Miguel, Edwin García, et al. 2018. Detection of Cyanuric Acid and Melamine in Infant Formula Powders by Mid-FTIR Spectroscopy and Multivariate Analysis. Journal of food quality, Volume 2018 Article ID 7926768
Ningrum, dian kartika, Gervacia Jenny Ratnawati, Indah Purwaningsih. 2018. Pengaruh Suhu Penyeduhan Terhadap Kadar Protein Pada Susu Formula Menggunakan Metode Kjeldahl. Jurnal Laboratorium Khatulistiwa e-ISSN : 2597-9531
Preechakasedkit, P., Kulwadee P., Wijitar D., Weena S., Wanpen immunochromatographic strip test using gold nanoparticles for the rapid detection of Salmonella typhi in human serum. Biosensors and Bioelectronics : 562– 566.
Wang, Zongyi, et al. 2010. Screening and Determination of Melamine Residues in Tissue and Body Fluid Samples. Analytica Chimica Acta 662 : 69–75.
Wei, DU Xin., ZHANG Yan-xin, SHE Yong-xin, LIU Guang-yang, ZHAO Feng-nian1, WANG Jing, WANG Shan-shan, JIN Fen, SHAO Hua, JIN M1zao-jun, ZHENG Lu-fei. 2016. Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics. Journal of Integrative Agriculture 2016, 15(5): 1166–1177
Yin, W., Liu, J., Zhang, T., Li, W., Liu, W., Meng, M., He, F., Wan, Y., Feng, C., Wang, S. 2010. Preparation of monoclonal antibody for melamine and development of an indirect competitive ELISA for melamine detection in raw milk, milk powder, and animal feeds. J. Agic. Food Chem., vol. 58 : 8152– 8157.
Zeng H., Ran Y., Qing-wen W., Jian-jun L., Ling-bo Q. 2011. Determination of Melamine by Flow Injection Analysis Based on Chemiluminescence System. Food Chemistry 127 : 842–846.
Zhou, Yu, et al. 2012. Monoclonal antibody based inhibition ELISA as a new tool for the analysis of melamine in milk and pet food samples. Food Chemistry Volume 135, Issue 4, 15 December 2012, Pages 2681-2686
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