The keto acids contain a carbonyl group on adjacent carbon atom to carboxylic acid in PYR KB MKBA KG K3MVA K4MVA and PPY and reacted with DDB to form cyclic ring structure as substituted tetramethyl dihydro-pyrazinol (Fig.
3 4) (Table 2) with average recovery was 95 90 85 90 82 88 and 90 for PYR KB MKBA KG K3MVA K4MVA and PPY (n=4) respectively.
In this case, the activity of PYR carboxylase was high (62.
Here, the activity of the PEP carboxylase (25) was half of the PYR carboxylase, while the activity of malic enzyme was zero.
In conclusion, hemolysis interference in urine and CSF is method dependent, and microhemolysis may contribute to unexpected urine and CSF protein results in laboratories using BTC and PYR
, whereas Hb does not seem to interfere in the BC method.
To an aqueous solution (1mL) containing GA, PYR, MKBA, KB, K3MVA, KG, K4MVA, KHA, and PPY of each within the concentration range as indicated in (Table 1), was added NPD reagent solution (1 mL, 1%, w/ v in methanol) and acetic acid-sodium acetate buffer pH 3 (1 mL).
A part was treated as HPLC analytical procedure and other was added PYR (3 ug), KB (8 ug), MKBA (6 ug), K3MVA (30 ug), KG (30 ug), K4MVA (60 ug), PPY (40 ug), and procedure HPLC was again followed the final volume was adjusted to 5 mL.
We compared the performance characteristics of our method with those of the Bradford CBB assay (7), the PYR assay (14), and the BTC assay (3).
The PYR assay was linear up to 3000 mg/L; the BC, BTC, and M-TP assays were linear up to 1500 mg/L; and the CBB assay was linear up to 1000 mg/L.
Finally, PYR, which does not bind to albumin, was not decreased in elderly subjects compared with adults 19-59 years of age.
5] Nonstandard abbreviations: BCKA, branched-chain keto-acid; KIC, [alpha]-ketoisocaproate; KIV, [alpha]-ketoisovalerate; KMV, [alpha]-keto-p-methylvalerate; PYR, pyruvate; KV, ketovalerate; OPD, o-phenylenediamine; and IS, internal standard.
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