Clear And Open Spirit Injection Hydrolyzed Buffalo Horn Quality Control And The Near Infrared Methodology Research

Qing Kai Ling injection is a traditional Chinese medicine injection which developed from AnGongNiuHuang Pill. With the complex prescription, Qing Kai Ling injection has significant curative effect, and is one of the national essential drugs. As the adverse reactions reported are increasing, the quality control method of Qing Kai Ling injection needs to be further improved.In the process of Qing Kai Ling injection production:Buffalo horn is hydrolyted to add to the product. The whole hydrolysis process of buffalo horn in the production of the Qing Kai Ling injection time was lasting for7-8h.So far there has not yet established the hydrolysis process and quality control methods of hydrolyzateand the chang trend of each active ingredient is not clear. So it needs to set a real-time online monitoring method to guarantee the stability and conformity of the hydrolysis process.In addition, near-infrared spectroscopy (NIR) technology is a fast, non-destructive analysis technics. With the rapid development of computer technology, near infrared spectroscopy is widely used in various fields. However, the parameters of traditional NIR analysis technics cannot evaluate the model stability and veracity. And near-infrared spectroscopy has still not formed a unified approach to methodology, which affects further development.To solve those problems above, this paper systematically studied the QingKaiLing Injection buffalo horn hydrolyzate quality control methods, and preliminarily discuss the establishment of near-infrared methodology. The main research contents and results are as follows:1The study of amino acids in Qing Kai Ling Injection and buffalo horn hydrolyteLiquid chromatography-mass spectrometry isotope labeling method was use to determine the type and content of amino acids in Qing kai ling injection and water buffalo horn hydrolyte. Then the phenyl isothiocyanate (PITC) pre-column derivatization reaction HPLC was establish to accurately measure content of12kinds of amino acids in water buffalo horn. The results showed that the establish method were accurate, laying a good foundation for the subsequent study of buffalo horn hydrolyzate quality control.2The analysis of hydrolysis process of buffalo horn in Qing Kai Ling InjectionTaking the12kinds of amino acids which have higher content levels in Buffalo horn as an indicator, the PITC pre-column derivatization high performance liquid chromatography method was used to analysis buffalo horn alkaline solution. The results showed that the content of serine decreased with the increase of hydrolysis time. The other11kinds of amino acid concentration increased until to the final equilibrium. But the balance time of12kinds of amino acids were different. The time of reaching balance for aspartic acid and glycine was shorter, at about100minutes, and for histidine was longer, at about500minutes. The hydrolysis process of buffalo horn was fitted by different kinds of kinetic equation by SAS statistical software. The results showed that the amino acid dissolution curve were in line with the first order equation, that can be used to describe hydrolysis process. The similarity of the hydrolysis process was analyzed by f2factor. The results showed that the amino acid hydrolysis curves were statistically significant.3. The NIR online quality control of the hydrolysis process of buffalo horn in Qing Kai Ling injectionTaking the12kinds of amino acids which have higher content levels in Buffalo horn as indicators, and the HPLC with Pre-column PITC Derivation as reference method, the quantitative calibration model between near-infrared spectral and HPLC reference values was established. The results showed that NIR can be used in the on-line quality control of the hydrolysis process of buffalo horn to except for asp, glu, gly and ser.4. The study of NIRS methodologyTaking baicalin of Shuanghuanglian oral liquid as the carrier, the quantitative calibration model between near-infrared spectral and HPLC reference values was established. The accuracy profile methodology was introduced to evaluate the NIR quantitative model, in order to overcoming the one-sidedness of traditional evaluation parameters R2. RMSEC, RMSEP. The results showed that:the accuracy profile can be used to evaluate the NIR quantitative model in Chinese medicine system.