title: Analysis of 26 amino acids in human plasma by HPLC using AQC as derivatizing agent and its application in metabolic laboratory.
abstract: The present study reports the simultaneous analysis of 26 physiological amino acids in plasma along with total cysteine and homocysteine by high-performance liquid chromatography (HPLC) employing 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) as precolumn derivatizing reagent. Separations were carried out using Lichrospher 100 RP-18e (5 μm) 250 × 4.0 mm column connected to 100 CN 4.0 × 4.0 mm guard column on a quaternary HPLC system and run time was 53 min. Linearity of the peak areas for different concentrations ranging from 2.5 to 100 pmol/μL of individual amino acids was determined. A good linearity (R (2) > 0.998) was achieved in the standard mixture for each amino acid. Recovery of amino acids incorporated at the time of derivatization ranged from 95 to 106 %. Using this method we have established the normative data of amino acids in plasma, the profile being comparable to the range reported in literature and identified cases of classical homocystinuria, cobalamin defect/deficiency, non-ketotic hyperglycinemia, hyperprolinemia, ketotic hyperglycinemia, urea cycle defect and maple syrup urine disease.
authors: Department of Pediatrics, Postgraduate Institute of Medical Education and Research, Chandigarh, 160012, India.
authors: Gaurav
authors: G
authors: Sharma
authors: Savita Verma
authors: SV
authors: Attri
authors: Bijaylaxmi
authors: B
authors: Behra
authors: Swapnil
authors: S
authors: Bhisikar
authors: Praveen
authors: P
authors: Kumar
authors: Minni
authors: M
authors: Tageja
authors: Sheetal
authors: S
authors: Sharda
authors: Pratibha
authors: P
authors: Singhi
authors: Sunit
authors: S
authors: Singhi
mesh_terms: D000596:Amino Acids / Q000097:blood* / Q000737:chemistry; D000634:Aminoquinolines / Q000737:chemistry*; D002219:Carbamates / Q000737:chemistry*; D002648:Child; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D002658:Developmental Disabilities / Q000097:blood; D005260:Female; D006712:Homocystinuria / Q000097:blood; D006801:Humans; D008297:Male
chemical_list: C078178:6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; D000596:Amino Acids; D000634:Aminoquinolines; D002219:Carbamates