Establishment and Validation of an New Immunity Chip Synthetic Method for High-Flux Determination of Ingredients in Chinese Materia Medica

Abstract

We have established a new immunity chip synthetic method for high-flux determination of the total ingredients within the Chinese Materia Medica (CMM) according to their labeled immunology [1,2]. In nature, the discrimination of heterogeneous ingredients among biologic systems is conducted through immunologic reactions between antigens and antibodies with specific and non-specific groups. We have taken the ingredients from CMM either directly as an antigen, or indirectly as a semi-antigen to be synthesized chemically, which was then injected into animals to produce the specific antibody to be made for an immunity chip, which could be used to obtain the cross reaction information for any antibodies to any semi-antigens. For the non-cross reaction ingredients, we could then read out its content directly according to the curve of labeled immunological competitive reaction. For the cross-reaction ingredients, we could carry out a labeled immunological competitive reaction on antibodies to semi-antigens in CMM and relevant formulas, then use a multivariant concentration or logarithm concentration linear matrix equation to resolve their concentrations. Taking these together we were able to establish an immunity chip synthetic method for the determination of ingredients in CMM's and formulas. With it we were able to analyze chrysophanol, emodin, emodin monomethyl ether, Rhein, Aloe-emodin in Radix and Rhizoma Rhei with a cross reaction ratio lower than 15%, and contents of 0.7521%, 0.6732%, 0.1483%, 1.239%, 0.5732%, with RSD of 2.967%, 6.471%, 5.257%, 4.878%, 3.551% respectively. The presented results will outline the newly developed immunity chip synthetic method in conjunction with HPLC determinations [3]. Acknowledgements: Thanks go to the Pharmacogenetic Research Institute, Central South University, Honghao Zhou, for partial support for this research.