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Abstrak :
[AFB1 merupakan mikotoksin yang dihasilkan jamur Aspergillus terutama A. flavus dan A. parasiticus yang hidup di daerah tropis. AFB1 dapat ditemukan pada berbagai produk pertanian dan makanan pokok mamalia seperti kacang tanah, jagung, kedelai, beras, dan gandum sehingga produk ini beresiko tinggi terkontaminasi toksin terutama AFB1 pada masa panen maupun pada masa penyimpanan. Resiko terbesar yang ditimbulkan toksin ini adalah kanker dan kematian. Telah banyak metode atau upaya yang dikembangkan untuk pendeteksian dini terhadap keberadaan jamur atau aflatoksin. Salah satu metode yang sederhana, terjangkau (murah), dan teruji kehandalannya adalah immunoassay dan immunokromatografi. Immunoassay membutuhkan antibodi sebagai pengenal AFB1. Pada penelitian ini antibodi AFB1 diproduksi. Mula-mula hapten AFB1 yang telah berhasil disintesis dikonjugasikan dengan protein (BSA) untuk dijadikan immunogen melalui metode ester aktif. Immunogen lalu disuntikan ke kelinci untuk menghasilkan antibodi poliklonal. Pembentukan antibodi terjadi pada hari ke-15 terhitung dari hari pertama imunisasi. Keberadaan antibodi dipastikan melalui Gel Agarose Precipitation Test (AGPT). Endapan yang terbentuk menunjukan terjadinya ikatan antara antibodi AFB1 dengan antigennya. Antibodi dimurnikan untuk dikonjugasikan dengan nano CdS hasil sintesis. Hasil konjugasi digunakan untuk mengembangkan biosensor berupa immunostrip pendeteksi aflatoksin B1. Metode immunostrip dijadikan alternatif deteksi karena teknik ini lebih praktis, cepat, mudah dilakukan oleh siapa saja.

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Aflatoxin B1 (AFB1), is mycotoxin produced by the fungus Aspergillus, especially A. flavus and A. parasiticus lived in the tropics. AFB1 composed by coumarin and two rings of furan. AFB1 can be found in a variety of agricultural products and basic food of mammals such as peanuts, corn, soybeans, rice, and wheat therefore they have high-risk products contaminated by toxin mainly AFB1 on the harvest or during storage. The greatest risk of the toxin is cancer or even death. Many methods have been developed for early detection of the presence of aflatoxins. One method that is simple, affordable (cheap), and proven reliability are immunoassay and immunochromatografi. Immunoassay needs antibody of AFB1 for the sensing antigen. The Immunogen synthesized by conjugating hapten of aflatoxin B1 with bovine serum albumin (BSA) by using the active ester method. The hapten – BSA was then injected into rabbits to produce polyclonal antibodies. The antibodies can be detected at the 15th days after the injection had been conducted. The presence of antibodies is confirmed using Agarose Gel Precipitation Test (AGPT). The precipitation showed the bond between AFB1 antibodies with antigens. The purified antibody then conjugated with nano CdS. The conjugate then applied to develop the biosensor for detection of AFB1 using immunostrip based on immunokromatographic method as an alternative method which can provide ease, rapid, simple method and practicable for everyone., Aflatoxin B1 (AFB1), is mycotoxin produced by the fungus Aspergillus, especially A. flavus and A. parasiticus lived in the tropics. AFB1 composed by coumarin and two rings of furan. AFB1 can be found in a variety of agricultural products and basic food of mammals such as peanuts, corn, soybeans, rice, and wheat therefore they have high-risk products contaminated by toxin mainly AFB1 on the harvest or during storage. The greatest risk of the toxin is cancer or even death. Many methods have been developed for early detection of the presence of aflatoxins. One method that is simple, affordable (cheap), and proven reliability are immunoassay and immunochromatografi. Immunoassay needs antibody of AFB1 for the sensing antigen. The Immunogen synthesized by conjugating hapten of aflatoxin B1 with bovine serum albumin (BSA) by using the active ester method. The hapten – BSA was then injected into rabbits to produce polyclonal antibodies. The antibodies can be detected at the 15th days after the injection had been conducted. The presence of antibodies is confirmed using Agarose Gel Precipitation Test (AGPT). The precipitation showed the bond between AFB1 antibodies with antigens. The purified antibody then conjugated with nano CdS. The conjugate then applied to develop the biosensor for detection of AFB1 using immunostrip based on immunokromatographic method as an alternative method which can provide ease, rapid, simple method and practicable for everyone]

Abstrak :
Kontaminasi aflatoksin pada makanan memberikan dampak yang sangat berbahaya bagi kesehatan. Kebijakan pemerintah harus diperketat dengan melakukan pemeriksaan pada setiap bahan pangan pokok. Dalam setiap pemeriksaan, baku aflatoksin diperlukan. Namun, keterbatasan anggaran untuk membeli baku tersebut menjadi hambatan pemerintah dalam melakukan pemeriksaan. Penelitian ini bertujuan untuk memproduksi bahan baku aflatoksin dengan metode yang lebih sederhana dan ekonomis sehingga dapat membantu pemerintah menyediakan baku aflatoksin. Konsentrasi aflatoksin tertinggi dihasilkan pada media kacang tanah sehingga digunakan dalam pembuatan baku aflatoksin. Waktu penyimpanan kacang tanah (2-4 minggu) dan pelarut untuk ekstraksi (metanol atau asetonitril) dilakukan optimasi. Penentuan kondisi analisis optimum dilakukan dengan membuat variasi komposisi fase gerak dan panjang gelombang emisi. Analisis dilakukan menggunakan KCKT Shimadzu® dengan detektor fluoresensi RF-10AXL, kolom C18 pada laju 0,8 mL/menit, suhu 40oC. Hasil optimasi kondisi analisis pada panjang gelombang emisi dan komposisi fase gerak masing-masing adalah 360 nm dengan air-metanol (60:40). Hasil validasi aflatoksin B2 diperoleh persamaan garis linier y = 5111,5x - 589,6 dengan nilai koefisien relasi (r) sebesar 0,9997. Hasil LOD dan LOQ yang didapatkan sebesar 0,6818 pg dan 2,2727 pg. %UPK dan %KV yang didapatkan sebesar 60-80% dan 0,3986-0,9545%. Waktu penyimpanan kacang tanah dan pelarut ekstraksi yang optimum adalah 4 minggu dengan pelarut metanol. Konsentrasi aflatoksin B2 dalam metanol yang didapatkan dari hasil penyimpanan 414,61 gram kacang tanah selama 4 minggu sebesar 79,74 ppb. pH telah diuji pada hasil produksi larutan aflatoksin B2 dan menunjukkan pH yang sesuai dengan kestabilannya. ......Aflatoxin contamination in foods cause very dangerous effect on health. Government policies must be tightened to do inspections on every staple food. In each inspection, aflatoxin standards are required. However, the limited budget for purchasing that standards is an obstacle for the government in conducting inspections. This study aims to produce aflatoxin raw materials with more simple and economical method so it can help the government to supply the standards. The highest aflatoxin concentration is produced in peanut media so it is chosen in making raw aflatoxin. The storage time of peanuts (2-4 weeks) and the solvent for extraction (methanol or acetonitrile) are optimized. The determination of optimum analysis conditions was also carried out by varying the composition of the mobile phase and emission wavelengths. Analyzes were performed using HPLC Shimadzu® with RF-10AXL fluorescence detector, C18 column at a rate of 0,8 mL/min, at 40oC. The results of the optimization of the analysis conditions of the emission wavelength and mobile phase composition are 360 nm with water-methanol (60:40). The results of validation of aflatoxin B2 were obtained linear regression y = 5111,5x-589,6 with correlation coefficient (r) 0,9997. The results of LOD and LOQ were 0,6818 pg and 2,2727 pg. %UPK and %KV were 60-80% and 0,3986-0,9545%. The optimum storage time for peanuts and extraction solvents is 4 weeks with methanol. The concentration of aflatoxin B2 in methanol obtained from 4 weeks of storage of 414,61 grams of peanuts for 79,74 ppb. pH has been tested on the result of the production of aflatoxin B2 solution and shown the pH in accordance with its stability.

Abstrak :
[ABSTRAK
Antibodi poliklonal anti aflatoksin B1 telah berhasil diproduksi pada hewan uji kelinci betina New Zealand White setelah diimunisasikan hapten aflatoksin B1-CMO yang dikonjugasikan dengan Bovine Serum Albumin (BSA) sebagai antigen. Hapten aflatoksin B1-CMO disintesis menggunakan metode karbodiimida dengan substrat aflatoksin B1 dan carboxymethyl hydroxylamine hemihydrochloride (CMO) sebagai linkernya. Hasil karakterisasi kromatografi lapis tipis dengan nilai Rf rata-rata sebesar 0.395, spektrum UV-Visibel dengan puncak λ maks pada 362, 264, 218 nm, spektrum IR dengan puncak 3448.126 cm-1 (3000-3600 cm-1) : OH, pada 1632.249 cm-1(1540-1725 cm-1) : C=O, dan 1642.451 cm-1 (1640-1690 cm-1) :C=N (Oksim), dan hasil fragmentasi spektrometri massa (MS/MS) pada m/z 386, 368.2, 310 membuktikan hapten aflatoksin B1-CMO berhasil disintesis. Hapten ini kemudian dikonjugasikan dengan BSA membentuk antigen aflatoksin B1-BSA (AFB1-BSA) sebelum diimunisasikan ke kelinci. Spesifitas antigen AFB1-BSA terhadap antibodinya dan uji konjugasi hapten ke BSA menunjukkan hasil positif menggunakan uji Dot Blot Immunoassay dengan konsentrasi BSA di dalam antigennya sebesar 1.74 mg/mL. Serum darah kelinci berdasarkan uji Agar Gel Precipitation Test (AGPT) positif mengandung antibodi poliklonal anti aflatoksin B1 setelah dua pekan (hari ke-11) sejak imunisasi primer antigen AFB1-BSA dilakukan. Dari serum darah bleeding panen, diperoleh konsentrasi antibodinya sebesar sebesar 2.19 mg/mL. Immunokromatogafi strip tes berhasil dibuat dengan nanopartikel iridium oksida (IrO2 NPs) sebagai kandidat label antibodinya dan dapat digunakan untuk mendeteksi sampel H IgG pada rentang 0.1 μg/mL sampai 10 μg/mL. Studi pendahuluan ini menunjukkan bahwa perangkat strip tes ini dapat digunakan untuk aplikasi konjugat sensor antibodi anti aflatoksin B1-nanopartikel iridium oksida untuk deteksi aflatoksin B1.

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ABSTRACT
Polyclonal antibody against aflatoxin B1 have been successfully produced in New Zealand White Rabbit after immunized by hapten of aflatoxin B1-CMO conjugated with Bovine Serum Albumin (BSA) as antigen. Hapten of aflatoxin B1-CMO was synthesized using carbodiimide method with afltoksin B1 as substrate and carboxymethyl hydroxylamine hemihydrochloride (CMO) as its linker. The characterization results of thin layer chromatography with Rf value of 0.395, the spectrum of UV-Visible with λ max peaks at 362, 264, 218 nm, the IR spectrum with peak at 3448.126 cm-1 (3000-3600 cm-1): OH , 1632.249 cm-1(1540-1725 cm-1): C = O, 1642.451 cm-1 (1640-1690 cm-1): C = N (oxime), and the results of mass spectrometry fragmentation (MS / MS) at m/ z of 386, 368.2, 310 proved that hapten of aflatoxin B1 -CMO successfully synthesized. Then, the hapten was conjugated to BSA to form antigen of aflatoxin B1-BSA (AFB1-BSA) before immunized to rabbits. The specificity of antigen of AFB1-BSA to its antibody and the confirmation of hapten-BSA conjugated showed positive results using dot blot immunoassay with BSA concentration in the antigen of 1.74 mg/mL. Based on Agar Gel Precipitation Test (AGPT) shown the rabbit blood serum resulted positive for polyclonal antibody against aflatoxin B1 after two weeks (day 11st) since the primary immunization of its antigen. From blood serum bleeding at harvest obtained the concentration of antibodies was 2.19 mg / mL. An Immunochromatogaphic test strip was successfully fabricated using iridium oxide nanoparticles (IrO2 NPs) as a labeled antibody candidate and can be used to detect the IgG H sample between of 0.1 μg/mL to 10 μg/mL. This preliminary study shown that the device can be used for applications of antibody against aflatoxin B1-nanoparticle iridium oxide conjugate for detection of aflatoxin B1 , Polyclonal antibody against aflatoxin B1 have been successfully produced in New Zealand White Rabbit after immunized by hapten of aflatoxin B1-CMO conjugated with Bovine Serum Albumin (BSA) as antigen. Hapten of aflatoxin B1-CMO was synthesized using carbodiimide method with afltoksin B1 as substrate and carboxymethyl hydroxylamine hemihydrochloride (CMO) as its linker. The characterization results of thin layer chromatography with Rf value of 0.395, the spectrum of UV-Visible with λ max peaks at 362, 264, 218 nm, the IR spectrum with peak at 3448.126 cm-1 (3000-3600 cm-1): OH , 1632.249 cm-1(1540-1725 cm-1): C = O, 1642.451 cm-1 (1640-1690 cm-1): C = N (oxime), and the results of mass spectrometry fragmentation (MS / MS) at m/ z of 386, 368.2, 310 proved that hapten of aflatoxin B1 -CMO successfully synthesized. Then, the hapten was conjugated to BSA to form antigen of aflatoxin B1-BSA (AFB1-BSA) before immunized to rabbits. The specificity of antigen of AFB1-BSA to its antibody and the confirmation of hapten-BSA conjugated showed positive results using dot blot immunoassay with BSA concentration in the antigen of 1.74 mg/mL. Based on Agar Gel Precipitation Test (AGPT) shown the rabbit blood serum resulted positive for polyclonal antibody against aflatoxin B1 after two weeks (day 11st) since the primary immunization of its antigen. From blood serum bleeding at harvest obtained the concentration of antibodies was 2.19 mg / mL. An Immunochromatogaphic test strip was successfully fabricated using iridium oxide nanoparticles (IrO2 NPs) as a labeled antibody candidate and can be used to detect the IgG H sample between of 0.1 μg/mL to 10 μg/mL. This preliminary study shown that the device can be used for applications of antibody against aflatoxin B1-nanoparticle iridium oxide conjugate for detection of aflatoxin B1 ]