| Dolutegravir is a heavyweight new anti-AIDS drugs developed by GSK,which has many advantages,such as high drug resistance,less drug interactions and superior efficacy,and has a promising market prospect in the future.R-3-aminobutanol,as a chiral amino alcohol compound,has been widely used in organic synthesis and drug production and it is the chiral source of the novel anti-AIDS drug,dolutegravir.This article focuses on the detection of 3-aminobutanol and the preparation of R-3-aminobutanol by a biological resolution method.In order to solve the limitations of the current 3-aminobutanol detection method,a new method for rapid and convenient detection of3-aminobutanol using charge transfer reaction and liquid chromatography was developed.The concept,features and application of charge-transfer complexes were introduced.Illustrate that the possibility of liquid phase detection of 3-aminobutanol without UV absorption.Through the investigation of buffer pH,loading reagent amount,reaction temperature and time,as well as the determination of liquid chromatography conditions including flow ratio,flow rate,column temperature,and gradient time.Concretely confirmed the charge-transfer reaction between aminobutanol and TCBQ were reacted completely at 60?water bath for 60 min under pH8.4 buffer solution.The determined liquid phase detection conditions were as follows:0.001%triethylamine aqueous solution-methanol gradient elution,column temperature 25°C,flow rate 1.0 mL·min-1,injection volume 10?L.The linear range of detection method for aminobutanol was0.1-0.6 g·L-1,R2=0.9994.The recovery rate of the method was98.3%-103.6%,and the RSD%was 0.205-0.344.The charge transfer reaction-HPLC method established in this study has high resolution and sensitivity,high repeatability of analysis results,simple operation,and is not easily affected by impurities and other factors.The detection range is wide,and it can be used for other non-UV absorption aliphatics amino alcohols.The determination of amino alcohols provides a powerful analytical tool for the detection and study of aminobutanol and the detection of other amino alcohols.A screening model of aminobutanol-degrading bacteria was established.Four strains of bacteria capable of degrading 3-aminobutanol were screened with mixed aminobutanol as the sole carbon source or the sole nitrogen source.The strains were identified as Streptomyces sp.?C1?,Paenibacillus sp.?C2?,Burkholderia sp.?C3?and Enterobacter sp.?N1?.The ability of four strains to degrade 3-aminobutanol and the optical configuration were analyzed by charge transfer reaction-HPLC and gas chromatography-derivatization methods.It was found that R-aminobutanol and S-aminobutanol consumed by C1 strains is almost the same,N1 strains consumed S type more than R type 4%,while C2 and C3 consumed S type more than R type 8%.In the future,the four strains will be further analyzed to analyze the mechanism and specific enzymes for degradation of3-aminobutanol,and the enzymes will be subjected to exogenous expression and orientation transformation.Combined with the optimization of culture conditions,the specific and efficient degradation of S-3-Aminobutanol will be performed.Thus,the purpose of preparing R-3-aminobutanol by bio-resolving method is to replace the existing high-cost chemical resolution method. |
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