ENKAPSULASI ANTIBODI ANTI OKRATOKSIN A DAN ANTI AFLATOKSIN B1 DALAM SILIKA GEL UNTUK CLEAN UP PADA ANALISIS OKRATOKSIN A DAN AFLATOKSIN B1 DALAM PALM KERNEL CAKE DENGAN HPLC-FLD; ENCAPSULATION OF ANTI-OCHRATOXIN A AND ANTI-AFLATOXIN B1 ANTIBODY ON SILICA GEL FOR CLEAN UP IN ANALYSIS OF PALM KERNEL CAKE OCHRATOXIN A AND AFLATOXIN B1 BY MEAN HPLC-FLD
PRANOWO, DENI, Nuryono
2016 | Tesis | FMIPAEncapsulation of anti-OTA and anti-AFB1 antibodies on silica gel has been done in order to perform clean up OTA and AFB1 analysis in Palm Kernel Cake (PKC) with HPLC-FLD. For this purpose, the antibodies were trapped in pore of silica gel, made of the TMOS (SiTMOS), commercial Na2SiO3 (SiKom) and Na2SiO3 extracted from rice husk ash (SiASP) as raw materials using sol gel technique. The modified silica gel with antibodies were then used as Immunoaffinity Chromatography Column (IAC) matrix and applied for OTA and AFB1 clean up performed in PKC. Matrix of IAC-SG OTA column was prepared by encapsulating of 1 mg Ab-OTA in 0.5 g silica gel of TMOS. While, for matrices of IAC-SG AFB1 SiTMOS, IAC-SG AFB1 SiKom, and IAC-SG AFB1 SiASP columns were prepared by encapsulating of 13.6 mg Ab-AFB1 in 0.5 g silica gel from SiTMOS, SiKom, and SiASP, respectively. In addition, the matrices were packed into the columns, followed by recovering and re-using tests prior being applied to the real samples. The recovery result of IAC-SG OTA was 95.71 ± 6.18%, the column could be used for eight times repetition with the recovery result reached more than 80%. Furthermore, the OTA in PKC using IAC-SG OTA clean up found 12 of 20 contaminated samples with the highest level of 1.47 ?g/kg and the average of 0.19 ?g/kg, while for using comercial IAC, found 13 of 20 contaminated samples with the highest level of 1.56 ?g/kg and the average of 0.21 ?g/kg. Both analytical results were linear correlated and not significantly different. The recovery results of IAC-SG AFB1 SiTMOS, IAC-SG AFB1 SiKom, and IACSG AFB1 SiASP columns of AFB1 were 92.12 ± 7.86, 93.75 ± 6.57, and 91.78 ± 9.50%, respectively. The binding capacity of the IAC-SG AFB1 SiTMOS columns was 40 ng, and for IAC-SG AFB1 SiKom and IAC-SG AFB1 SiASP columns were 50 ng. All the columns could be re-used up to five times repetitions with recovery more than 80%. In addition, based on PKC analysis, 9 of 10 AFB1 contaminated samples were found. The highest level of AFB1 found after clean up using IAC-SG AFB1 SiTMOS was 6.83 ?g/kg with average of 1.14 ?g/kg, while for IAC-SG AFB1 SiKom and IAC-SG AFB1 SiASP samples were 7.59 ?g/kg with average of 1.56 ?g/kg, and 8.03 ?g/kg with average of 1.73 ?g/kg, respectively. Whereas for the commercial IAC, the highest level of clean up was reached 9.49 ?g/kg and the average was 1.62 ?g/kg. The clean up recovery result using IAC comercial is higher in comparison with the other colums, however they were linear correlated.
Kata Kunci : N
