Separation of free amino acids in human plasma by capillary electrophoresis with laser induced fluorescence: potential for emergency diagnosis of inborn errors of metabolism

Boulat, Olivier; McLaren, David G.; Arriaga, Edgar A.; Chen, David D. Y.

doi:10.1016/s0378-4347(00)00611-3

Cited by 60 publications

(54 citation statements)

References 23 publications

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“…Since its introduction in the early 1980s [1] capillary electrophoresis (CE) has proven to be an invaluable tool for the separation of a wide variety of compounds, including those of biological [2,3], pharmaceutical [4] and industrial [5] interest. However, despite the widespread use of CE as a separation technique, there is still at least one area in which it is consistently outperformed by HPLC or slabgel electrophoresis: preparative operation.…”

Section: Introductionmentioning

confidence: 99%

A quantitative study of continuous flow‐counterbalanced capillary electrophoresis for sample purification

McLaren

Chen

2003

Electrophoresis

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A systematic evaluation of continuous flow-counterbalanced capillary electrophoresis for microscale preparative applications is presented. The success of the technique was found to depend on the specific nature of the analyte and background electrolyte and was most heavily influenced by factors such as solution conductivity and rate of buffer depletion. The performance of the technique for the system studied was ultimately limited by contamination arising from changes in analyte mobilities during extended run times. Marked improvements in both purification rate and yield were observed using the zwitterionic buffer 2-(N-cyclohexylamino)ethanesulfonic acid when compared to similar runs carried out using a borate background electrolyte. A quantitative evaluation of the technique was performed for the analytes studied and the performance of the technique has been compared to fraction collection methods. The highest recovery achieved was 5.2% of the fastest migrating component present in the original sample with a resolution of 9 to the adjacent peak and required 100 min under optimized conditions. Recovery could be further increased to 9.0% in 120 min for the same analyte using pressure-ramped flow-counterbalanced capillary electrophoresis.

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Section: Introductionmentioning

confidence: 99%

A quantitative study of continuous flow‐counterbalanced capillary electrophoresis for sample purification

McLaren

Chen

2003

Electrophoresis

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“…The separated analytes are commonly detected by an on-column UV-absorbance or laser-induced fluorescence (LIF) detectors. Several protocols have been developed for determination of AAs [32][33][34] and organic acids [35][36][37][38][39][40] such as orotic acid [41][42][43] in urine or blood samples using CE. Direct measurements can often be performed on samples with minimum initial pretreatment -i.e., centrifugation and dilution, if needed.…”

Section: Capillary Electrophoresismentioning

confidence: 99%

“…Such an arrangement is suitable for the analysis of substances with native fluorescence or the ones which can be readily derivatized. For example, the free AAs in human plasma were analyzed with high sensitivity after derivatization with 3-(4-carboxybenzoyl)quinoline-2-carboxylaldehyde (CBQCA) [32]. The derivatization step takes approximately 40 min with the analysis concentration limit of detection as low as 4610 28 M. Labeled AAs can be easily excited by the 488 nm line of an Ar-ion laser, or the 442 nm line of He/Cd laser.…”

Section: Capillary Electrophoresismentioning

confidence: 99%

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Capillary electrophoresis and mass spectrometry for screening of metabolic disorders in newborns

2004

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Clinical analyses always represent a challenge for the sensitivity and selectivity of the analytical techniques. Of the most critical are the techniques required for the quick determination of the disease state and application of the proper treatment in newborns. This short critical review overviews the present state of the art of the use of mass spectrometry and capillary electrophoresis for screening of metabolic disorders in newborns.

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“…To overcome the problem of detection, the determination of AA is generally accomplished by a combination of off-line derivatization and separation with CE [1][2][3][4][5][6]. It results in the loss of target compositions due to many transfers of very limited samples in the off-line manipulation.…”

Section: Introductionmentioning

confidence: 99%

On‐column labeling technique and chiral CE of amino acids with mixed chiral selectors and UV detection

Han

Chen

2007

Electrophoresis

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A novel method has been developed for the on-column labeling of amino acid enantiomers with 9-fluoroenylmethyl chloroformate (FMOC), followed by chiral CE with a binary chiral selector system and UV detection. Efficient labeling was achieved by sequential injection of amino acids, borate buffer, and FMOC labeling solution at 0.2 psi for 6 s. After injection, the sandwich sections were electrically mixed at 250 V/cm for 6 s and allowed to react (electric field-free) at room temperature for 2 min. With this procedure, successful online-labeling and chiral CE separation of 19 pairs of amino acids (AA) have been conducted, giving 17 pairs fully enantioresolved (R(s) = 1.73-5.79) and two pairs partially resolved (Ala, R(s) = 0.39 and Arg, R(s) = 1.15) using a running buffer of 150 mM borate containing 30 mM beta-CD, 30 mM sodium taurodeoxycholate (STDC), and 15% isopropanol (IPA) at pH 9.0. Chiral CE of some mixed pairs was also demonstrated, much the same as using precolumn labeling. Surprisingly, Met, Asp, Asn, Gln, and His gained even higher enantioresolution (up to 2.5%) compared with the case of precolumn labeling. As validated by both artificially prepared solutions and serum samples, the method was applicable to the quantitative determination of AA, with LODs down to 4.0 microM. The method allowed the determination of D-AA at the ratio of 1:100 (D:L).

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Separation of free amino acids in human plasma by capillary electrophoresis with laser induced fluorescence: potential for emergency diagnosis of inborn errors of metabolism

Cited by 60 publications

References 23 publications

A quantitative study of continuous flow‐counterbalanced capillary electrophoresis for sample purification

A quantitative study of continuous flow‐counterbalanced capillary electrophoresis for sample purification

Capillary electrophoresis and mass spectrometry for screening of metabolic disorders in newborns

On‐column labeling technique and chiral CE of amino acids with mixed chiral selectors and UV detection

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