Analysis Of The Separation Of Diaminopentane In Fermentation Broth

Diaminopentane (also known as1,5-diaminopentane, cadaverine) has attracted moreand more attention as a kind of platform compound due to the development of bio-economy.As a very important chemical products which have been widely used in industrialapplications, diaminopentane monomer has been largely adopted in various polyamides,polyurethanes, chelating agents and additives. Large scale production of diaminopentane byusing biotechnology is an alternative to the traditionally developed biogenicamines-hexamethylene diamine. Moreover, the dibasic acid polymerization reaction can beused to synthesize an organic polymer materials, a new type of nylon, which represents anenvironmentally friendly, sustainable model, for broad applications.Fermentation is new route to produce diaminopentane with improved production rate.However, this novel synthetic route is largely limited due to the difficulty of extract, refineand analyze diaminopentane from the complex composition of fermentation broth. In thispaper, we developed new methods to analyze diaminopentane from the fermentation brothby understanding the research status of the source, extraction, refining, quantitative andqualitative analysis method of diaminopentane. Pre-Column Derivation HPLC-UV andARPLC-ESI/MS were used as derivatives and non-derivative analysis method to analyzepentanediamine in fermentation broth quantitatively and qualitatively, respectively.The method of Pre-Column Derivation HPLC-UV was developed for determiningdiaminopentane in fermentation broth. By using dansyl chloride as the derivative reagent,the reaction only needs just10min with the assistance of ultrasonic wave. Different factorsthat affecting the derivatization were optimized to achieve the best results. The derivativeswere separated on a Hyersil BDS C18(2.1mm×100mm,3μm) with methanol-water(80:20, v/v) as mobile phase. The flow rate was0.2ml/min and the temperature of columnwas30℃. The column eluate was monitored by ultraviolet detector with a wavelength of254nm and quantified by external standard method. The analysis time for the sample wasless than6min, which effectively decrease the consumption of the mobile phase andconsequently save the cost of operation. Inthe concentrationrange from1to250μg/ml, thediaminopentane presented good linear correlation (R2>0.999) with the limit of detection 0.281ppb. Therefore, this devoloped method is a simple, fast, sensitive, reproducible andpractical strategy for the analysis of diaminopentane in fermentation broth.The other new method was based on the successful separation by using acidic reversedphase liquid chromatography (ARPLC) and followed by the detection via electrosprayionization tandem mass spectrometry (ESI/MS). YMC-Triart C18(2.0mm×100mm,3μm) chromatographic column was used as the stationary phase. It has excellent acidic andalkali resistance in a wide pH range from1to12and is suitable for the separation ofdiaminopentane. Moreover,10%2mM NH4Ac-0.1%HCOOH buffer solution and90%methanol were used as the mobile phase. The sample injection volume was2μl, the columntemperature was25℃, and the flow rate was150μl/min. The detection limit was37.42ppb and the analysis time was only1.22min. In addition, we also designed HILIC-ESI/MSas a analyze method to analysis diaminopentane. It was found that the retention time was8.39min, which was longer than that observed with ARPLC-ESI/MS. This should beattributed to the strong polarity of diaminopentane.