Detect Macromolecules In Traditional Chinese Medicine Injection And Their Cellular Toxicity

Objective:The main objective of this study was to prove the safety of Traditional Chinese Medicine injection(TCMI) is improved by detecting the macromolecules including polysaccharide, resin, protein and tannin, and the cellular toxicity. Four TCMIs(Qingkailing injection, Shuanghuanglian injection Danshen injection and Dengzhanxixin injection) were used as examples in the study.Method:(1) Detected polysaccharides in TCMIs by colorimetric method based on the principle that the polysaccharide is able to make the aniline blue fade.(2) Macromolecule-rich injections(retained fluid) and Macromolecule-free injections(filtrate) were prepared by molecular sieves with different pore diameters based on the principle of molecular sieve. Macromolecule-free injections were used to detect resin with an adapted method similar to Chinese Pharmacopoeia.(3) Macromolecule-free injections(filtrate) were used to prove the validity of the detection methods of protein and tannin, established by Wei-gang Duan.(4) The Cell Counting Kit-8(CCK-8) was used to observe the cellular toxicity both for macromolecule-rich injections and Macromolecule-free injections.Results:(1) Although aniline blue fade method can not accurately detect polysaccharide contented in TCMIs, but can indirectly reflect the amount of polysaccharide and protein, therefore, provides some help to improve the safety of TCMIs. The results showed that TCMI free of macromolecules contained less protein and polysaccharide.(2) A new and better method for the detection of resin was established by adapting the method in Chinese Pharmacopoeia, and resin contained in the injections was detected by this method. And the results showed that TCMI free of macromolecules contained less resin.(3) Feasibility and sensitivity of the detection methods of protein and condensed tannin were proved by used macromolecule-free injections(filtrate)prepared by molecular sieves with different pore diameters based on the principle of molecular sieve. And the results showed that TCMI free of macromolecules really contained less protein and condensed tannin.(4) Macromolecule-rich injections showed higher cellular toxicity, compared with macromolecule-free injections, which proved that the safety of TCMIs was enhanced by removing macromolecules.Conclusion:TCMI free of macromolecules made with molecular sieves contains less macromolecular substances, and has smaller cellular toxicity. All the results suggest that TCMI free of macromolecules is much safer.