Homocysteine is a non-essential sulfur containing amino acid closely linked to the essential amino acid methionine. Homocysteine is rapidly oxidized in plasma into homocystine and cysteinehomocysteine disulfide. Total plasma homocysteine is the sum of homocysteine and its oxidized forms. The determination of total homocysteine plays an indicator role in diagnosis and therapy of folate, vitamin B12 and vitamin B6 deficiencies. In addition, several observational studies have confirmed that increased total homocysteine is an independent predictor of cardiovascular risk.
Pipette 50 µL of serum/plasma or quality control sample into a glass centrifuge tube and add
50 µL of internal standard
Vortex for 5 sec. and add 100 µL of reducing solution, again vortex for 15 sec. Then,
incubate in a water bath at 60oC for 15 min.
Add 650 µL Reagent-2 and vortex for 5 sec. then centrifuge at 3000 rpm for 5 min.
Decant the clear supernatant into HPLC vial prior to injection