1) A. S. Sadykov Institute of Bioorganic Chemistry, Academy of Sciences of the Republic of Uzbekistan, 700143, Tashkent, ul. Kh. Abdullaeva, 83, fax (371) 162 70 71, e-mail: root@ibc.edu.uz; 2) University of Northern Colorado, 501 20 Street, Greeley, CO 80639, USA, e-mail: spmacke@bentley.unco.edu. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 482-485, November-December, 2001. Original article submitted July 24, 2001. 0009-3130/01/3706-0562$25.00 2001 Plenum Publishing Corporation © 562 Chemistry of Natural Compounds, Vol. 37, No. 6, 2001 COMPARATIVE BIOCHEMISTRY OF THE PHYSIOLOGICALLY ACTIVE COMPONENTS OF VENOM, HEMOLYMPH, AND EGGS OF THE KARAKURT SPIDER (Latrodectus tredecimguttatus)* A. A. Akhunov, Z. Golubenko, N. A. Abdurashidova, UDC 577.993 1 1 1 E. Ch. Mustakimova, F. A. Ibragimov, and S. Mackessy 1 1 2 The physicochemical and functional properties of the venom, hemolymph, and egg extract of the karakurt spider Latrodectus tredecimguttatus were studied. A highly purified toxin that differs from the neurotoxins found previously in the venom was isolated from the hemolymph and characterized. Key words: whole venom, hemolymph, karakurt spider egg extract. The use of biologically active natural products to solve scientific and technical problems of modern biology and medicine is a promising approach. Therefore, further searches for new physiologically active components in natural substances involve their identification, isolation, overall characteristics, and the determination of the areas and methods of application. These searches are important not only for fundamental science, which expands and deepens understanding of the environment, but also for practical application. Extracts of the venom glands of Latrodectus spiders have been used as starting materials to study the venom and latrotoxins in them [1]. Information about the toxic properties of hemolymph and egg extract was also obtained [2]. However, these latter studies were incomplete. Therefore, many issues are unresolved. The main ones concern the nature and properties of toxins in these materials and the possibility that they are identical or similar to toxins in the venomous secretions of these spiders. We present results from research on the physicochemical and functional properties of the venom, hemolymph, and egg extract of Latrodectus tredecimguttatus. As a rule, preparations obtained from the venom glands of the spiders contain a trace impurity of hemolymph components. Therefore, the study of these preparations using a commercial antiserum against karakurt venom that was prepared in Tashkent at a Scientific-Research Institute of the Ministry of Health of the Republic of Uzbekistan and monospecific antisera against α- and β-latrotoxins is of interest. Thus, antiserum with a 1:64 titer against the venom extracted from spider venom glands gives three precipitation lines upon binary radial immunodiffusion against spider hemolymph. Monospecific antisera against α-latrotoxin purified until homogeneous with a 1:64 titer and β-latrotoxin with a 1:32 titer gave no precipitation lines upon Ouchterlony reaction against hemolymph and egg extract. Therefore, hemolymph and egg extract contain toxins different from those of spider venom glands. A determination of the toxicity is the first step in the study of the biological activity of any natural substance. The toxicity (LD ) is known to depend on the physiological state of the specimens from which the venom is obtained, the time of 50 year, and the methods of preparation, drying, and storage, in addition to the geographical distribution of the population of venomous subjects. Furthermore, the sensitivity of experimental animal species to the sample is important [1]. We used venom and hemolymph of spiders collected in summer (July-August) in Dzhizak District of Uzbekistan. Spider egg silk was obtained in September from the insectarium of the Zoology Institute of the Academy of Sciences of the Republic of Uzbekistan. ______ *Present at the IVth International Symposium on the Chemistry of Natural Compounds (Isparta, Turkey, June 6-8, 2001).