KETO ACIDS Dotaz Zobrazit nápovědu
Annals of the New York Academy of Sciences ; vol. 573, no. 1, 1989
[1st ed.] XVIII, 474 s. : obr., tab., grafy ; 23 cm
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- biochemie
- MeSH
- chronické selhání ledvin dietoterapie krev moč MeSH
- dospělí MeSH
- esenciální aminokyseliny MeSH
- finanční podpora výzkumu jako téma MeSH
- hodnoty glomerulární filtrace MeSH
- lidé středního věku MeSH
- lidé MeSH
- metabolická clearance MeSH
- nízkoproteinová dieta MeSH
- senioři MeSH
- větvené aminokyseliny analýza metabolismus terapeutické užití MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- srovnávací studie MeSH
The determination of α-keto acids derived from amino acids is currently the most reliable approach for the diagnosis of some congenital metabolic diseases. An HPLC method for the simultaneous measurement of selected α-keto acids in dried blood samples has been developed and evaluated. Blood spot samples from a group of healthy blood donors were collected onto #903 Specimen Collection Paper. Prior the separation, the α-keto acids were derivatized with 1,2-diamino-4,5-dimethoxybenzene to the corresponding 3-substituted-6,7-dimethoxy-2(1 H)-quinoxalinol derivatives. For the separation, a reverse-phase column LichroCart 125-4, Purospher RP-18e, 5 μm, was used. The mixture of 25% ACN in deionized water (mobile phase A) and 100% ACN (mobile phase B) were used for a gradient elution of α-keto acids derivatives. Analytical performance of this method is satisfactory for all α-keto acids. The intra-assay and inter-assay coefficients were below 10% and recoveries were close to 100%. We have developed relatively simple, rapid, selective and sufficiently sensitive HPLC method with fluorescence detection for the determination of selected α-keto acids in dried blood samples. The presented method is suitable for clinical testing purposes.
- MeSH
- diabetická ketoacidóza etiologie farmakoterapie komplikace MeSH
- dítě MeSH
- draslík terapeutické užití MeSH
- edém mozku farmakoterapie MeSH
- fosfor terapeutické užití MeSH
- hydrogenuhličitan sodný terapeutické užití MeSH
- inzulin aplikace a dávkování terapeutické užití MeSH
- lidé MeSH
- rehydratační roztoky terapeutické užití MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- MeSH
- chronické selhání ledvin dietoterapie MeSH
- ketokyseliny aplikace a dávkování metabolismus MeSH
- lidé MeSH
- nízkoproteinová dieta metody využití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- ketokyseliny krev MeSH
- krevní nemoci diagnóza MeSH
- nemoci drůbeže diagnóza etiologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
BACKGROUND: The determination of branched chain amino acids [BCAA; valine (Val), leucine (Leu), isoleucine (Ile)], alpha-keto acids derived from BCAA [BCKA; alpha-ketoisovaleric acid (KIV), alpha-ketoisocaproic acid (KIC), alpha-ketomethylvaleric acid (KMV)], methionine (Met), phenylalanine (Phe) and tyrosine (Tyr) is currently the most reliable approach for the diagnosis of maple syrup urine disease (MSUD), hypermethioninemia, phenylketonuria (PKU) and tyrosinemia. The aim of this study was to develop rapid and simple HPLC methods for measurement of BCAA, Met, Phe, Tyr and BCKA in plasma and dried blood samples. METHODS: Samples of peripheral venous blood with EDTA as anticoagulant were obtained from a group of healthy blood donors (n=70, 35 females, 27-41 years of age and 35 males, 28-43 years of age). Blood-spot samples from a group of newborns (n=80, 40 girls and 40 boys 3-5 days of age) were collected onto #903 Specimen Collection Paper and allowed to dry for at least 24 h before analysis. Prior to separation, the amino acids (AA) were derivatized with o-phthaldialdehyde (OPA) and BCKA with o-phenylenediamine (OPD). Reverse phase column chromatography (LiChroCart 125-4 Purospher RP-18e, 5 microm) was used for separation and fluorescence detection used to monitoring of effluent. For AA analysis, 25 mmol/L sodium hydrogenphosphate-methanol (90:10, v/v), pH 6.5+/-0.1 was used as mobile phase A and 100% methanol was used as mobile phase B. Measurement of BCKA used a mixture of methanol and deionized water (55:45, v/v) as mobile phase A and mobile phase B consisted of 100% methanol. RESULTS: Analytical performance of these methods was satisfactory for the determination of all AA and BCKA. The intra-assay and inter-assay coefficients of variation were below 10% and recovery ranged from 90%-110%. CONCLUSIONS: We have developed simple, rapid and selective HPLC methods with fluorescence detection for the determination of BCAA, Met, Phe, Tyr and BCKA in plasma and dried blood samples.
- MeSH
- dospělí MeSH
- fenylalanin krev MeSH
- fluorescence MeSH
- ketokyseliny krev MeSH
- krevní skvrny MeSH
- lidé MeSH
- methionin krev MeSH
- novorozenec MeSH
- referenční hodnoty MeSH
- tyrosin krev MeSH
- větvené aminokyseliny krev MeSH
- vrozené poruchy metabolismu aminokyselin krev MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- novorozenec MeSH
- ženské pohlaví MeSH
