Journal of Chromatography & Separation Techniques
Open Access

Synthesis of two new derivatizing reagents and their application to separation of chiral drug

4^th World Congress on Chromatography

August 07-09, 2017 | Rome, Italy

Vinod Kumar Vashistha and Ravi Bhushan

GLA University, India
Indian Institute of Technology Roorkee, India

Posters & Accepted Abstracts: J Chromatogr Sep Tech

Abstract :

Under appropriate reaction conditions of reaction time, temperature and solvent, the chlorine atoms in cyanuric chloride (2,4,6â�? trichloroâ�?1,3,5â�?triazine; CC) can be substituted by nucleophiles in a controlled sequential manner, resulted into MCT (monochloro triazine) and DCT (dichloro triazine) based CDRs. Commercial availability and economic suitability makes the CC a more attractive starting material for the synthesis of CDRs. By these characteristics, synthesis of two MCT reagents, namely, Nâ�? (4â�?chloroâ�?6â�?piperidinylâ�?[1,3,5]â�? triazineâ�?2â�?yl)â�?Lâ�?Isoleucine; (CDRâ�?1) and Nâ�?(4â�?chloroâ�?6â�?piperidinylâ�?[1,3,5]â�? triazineâ�?2â�?yl)â�?LMethionine; (CDRâ�?2) were carried out by nucleophilic substitution of one of the chlorine atoms by a piperidinyl group and the second with L-amino acids (as the chiral auxiliary). These reagents were characterized and used for derivatization of (RS)-isoprenaline (spiked in human plasma). The diastereomeric derivative were separated on a reversed-phase C18 column with a mobile phase consist of acetonitrile and 0.1% TFA under gradient mode from 35-65% of acetonitrile at a flow rate of 1.0 mL min-1 andï�? UV detection at 254 nm. The method was validation according to ICH guidelines. The separation mechanism and elution order of the diastereomeric derivative were proposed and supported by developing the geometry optimized lowest energy structures of the two diastereomers using a DFT based program, Gaussian 09 Rev. A.02 and hybrid density functional B3LYP with 6-31G basis set. In L-(R)-diastereomer, the bulky moieties, alkyl group on the stereogenic center of amino acid (present in the CDR), and phenyl group on stereogenic center of Ipn, are oriented on the same side with respect to triazine moiety, i.e., cis orientation. In L-(S)-diastereomer, these groups are oriented in a manner anti to each other with respect to triazine moiety and thus have a trans-type arrangement. Therefore, being less hydrophobic, it is L-(S)-diastereomer which eluted before its counterpart.

Biography :

Email: vinod.vashistha@gla.ac.in