| 摘要 |
The invention relates to clinical biochemistry, biotechnology and medicine, in particular to a method of detecting permissible tralkoxydim doses and derivatives thereof in humans and animals based on reproduction in vitro lipolytic components of digestion action and can be used for pre-testing toxicity of novel biologically active compounds having pesticide activity. According to the invention a method is disclosed for detecting permissible tralkoxydim doses for humans and animals based on modeling digestion conditions characterized in detecting change of activity of pancreatic phospholipase Aby determining the content of phospholipase metabolites involving methemoglobin in kinetic regime using difference spectophotometry (AD) and including: a) preparation of first check sample comprising micellae of phosphatidylcholine of egg yolk or natural phosphatidylserine emulsified by bile acid with ratio phospholipid/bile acid 1:2-3 in buffer solution at pH 7.4-8.2 in the presence of CaCland solution of methemoglobin in tris-HCl buffer, preparation of second test sample comprising phospholipase Aand micellae of phosphatidylcholine of egg yolk or natural phosphatidylserine emulsified by bile acid in buffer solution at pH 7.4-8.2 in the presence of CaCland solution of methemoglobin in tris-HCl buffer, and recording the difference spectophotometry (AD) of said samples; b) preparation of third test sample pesticide being tested in suitable solvent, phospholipase A, micellae of phosphatidylcholine of egg yolk or natural phosphatidylserine emulsified by bile acid in buffer solution at pH 7.4-8.2 in the presence of CaCland solution of methemoglobin in tris-HCl buffer, and forth test sample comprising said solvent and, if necessary pesticide being tested, micellae of phosphatidylcholine of egg yolk or natural phosphatidylserine emulsified by bile acid in buffer solution at pH 7.4-8.2 in the presence of CaCland solution of methemoglobin in tris-HCl buffer, and recording the difference spectophotometry (AD) of said samples; c) subsequent calculation of change of the enzyme activity level established in the control by known arts and determining pesticide as safety provided the change of the enzyme level in its presence within at least 20% of the control. A reactant or a preparative pesticide form can be used as a compound to be tested, 3α,12α-di- and 3α,7α,12α-trihydroxycholanic acids (deoxycholic and cholic acids correspondingly). The use of phospholipotic reaction as a simple model of a process for destruction of food phospholipids provides to reliably evaluate pesticide safety for organisms of humans and animals. The inventive express-method has an obvious advantage since there is no need to perform durable tests using laboratory animals and creates favourable conditions for determining pesticides of new generations among a wide range of plants protection means safety for a human. |
