Development of in-house ELISA for detection of chloramphenicol in bovine milk with subsequent confirmatory analysis by LC-MS/MS

This study was undertaken to develop and validate direct competitive ELISA for the determination of chloramphenicol residues in bovine milk. Antisera and an enzyme-tracer for chloramphenicol were prepared and used to develop an ELISA with inhibition concentrations, IC₂₀ and IC₅₀, of 0.09 and 0.44 ng mL^?1, respectively. Milk samples were spiked with standards equivalent to 0, 0.2, 0.3, 0.5, 1.0 &; 1.5 ng mL^?1 and extracted in methanol. The mean recoveries were found to be 73–100% with coefficient of variance 7–11%. The decision limit (CCα) and detection capability (CCβ) were calculated as 0.10 and 0.12 ng mL^?1, respectively. The results were found comparable with the commercial ELISA, having recoveries of 87 to 100%, CCα 0.09 ng mL^?1 and CCβ 0.12 ng mL^?1. As per Commission Decision 2002/657/EC, in-house ELISA was further validated by using LC-MS/MS. Mass spectral acquisition was done by using electrospray ionization in the negative ion mode applying single reaction monitoring of the diagnostic transition reaction for CAP (m/z 152, 194 and 257). The calibration curve showed good linearity in concentrations from 0.025 to 1.6 ng mL^?1 with correction coefficient 0.9902. The mean recoveries were found to be 88 to 100%. The CCα was calculated as 0.057 ng mL^?1 and CCβ 0.10 ng mL^?1. Since CCα and CCβ are less than half of the MRPL (0.15 ng mL^?1), the test was found suitable for screening and quantification of CAP residues in bovine milk samples. Results of surveillance studies indicated that out of 31 analyzed milk samples, 12.9% samples were found with CAP residues but only 3.2% samples were declared positive with maximum concentration 0.31 ng mL^?1, slightly above the MRPL.     

Health risk assessment of hexachlorocyclohexane in soil,water and plants in the agricultural area of Potohar region,Punjab, Pakistan

Environmental Geochemistry and Health - As the largest organ of the human body, the skin is the major exposure route of NO2. However, the evidence for a relationship between NO2 exposure and...     

Role of NADH-dependent chromium reductases,exopolysaccharides and antioxidants by Paenibacillus thiaminolyticus PS 5 against damage induced by reactive oxygen species

ABSTRACT

Cr (VI) being used in various activities of industries and its improper treatment lead to contamination of environment. Among different methods, biological is the most efficient method to control pollution from soils affected with metals. Present study was designed to assess the role of Paenibacillus thiaminolyticus PS5 for adsorption, Cr (VI) reduction and mechanism of reduction. Sorption of chromium (VI) by strain PS5 was obtained by batch equilibrium method. Cr (VI) reduction in both free and immobilised cells were evaluated by 1,5-Diphenyl Carbazide method. The formation of biofilm by Paenibacillus thiaminolyticus PS5 was observed for colour change as well as quantified spectrophotometrically. Analysis kits were used to measure the amount of eDNA, superoxide and malondialdehyde (MDA). Metal resistant strain PS5 was characterised as P. thiaminolyticus using 16S rRNA gene sequence. Maximum biosorption of Cr (VI) by strain PS5 was found at pH 6–8 and 100–400 µgCr/mL within 24 hours of incubation. Complete reduction of Cr (VI) by strain PS5 was achieved at pH 6–8 and100–300 µg/mL Cr (VI). Paenibacillus thiaminolyticus PS5 immmobilisation on sodium alginate and polyvinyl alcohol facilitated complete reduction of Cr (VI) within 18 hours due to the formation of more biofilm under metal stress conditions. Strain PS5 reduced almost all Cr (VI) into Cr (III) in supernatant, most of which was immobilised by cell debris. Experiments confirmed the reduction of Cr (VI) by cytosolic cell-free extracts which is a mechanism of detoxification. The release of exopolysaccharides and antioxidants by strain PS5 played a protective role against cell damage by Cr (VI) as Cr (VI) could not release the significant amount of eDNA, superoxide and MDA. The present study proved strain PS5 to be a super bioinoculant which has great capacity for adsorption, biotransformation and can activate cytosolic reductases, exopolysaccharides and antioxidants against oxidative damage.     

Role of ambient air pollution in asthma spread among various population groups of Lahore City: a case study

Environmental Science and Pollution Research - Air pollution levels rise as a result of industrial and vehicular emissions, epidemiological issues such as asthma become more prevalent in Lahore,...     

Organochlorine pesticides in the surrounding soils of POPs destruction facility: source fingerprinting,human health,and ecological risks assessment

Environmental Science and Pollution Research - The elimination of persistent organic pollutants (POPs) obsolete pesticides stockpiles, particularly the organochlorine pesticides (OCPs), is one of...