Determination of lipoic acid in human plasma by high-performance liquid chromatography with ultraviolet detection

Chwatko, Grażyna; Krawczyk, Marta; Iciek, Małgorzata; Kamińska, Adrianna; Bilska‐Wilkosz, Anna; Marcykiewicz, Bernadeta; Głowacki, Rafał

doi:10.1016/j.arabjc.2016.10.006

Cited by 17 publications

(20 citation statements)

References 28 publications

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“…It indicates the need to control the concentration of each of these compounds. Although many analytical protocols dedicated to the determination of LA or metabolically related amino thiols in human plasma have been presented in the literature, all of them require the determination of LA and GSH, Cys, Hcy in two different assays [21][22][23][24][25][26][27][28][29][30].…”

Section: Resultsmentioning

confidence: 99%

“…To avoid these troubles, usually deproteinization with the use of The high separation capacity of high performance liquid chromatography (HPLC) makes it the preferred technique for biological samples analysis. For quantification of LA in biological fluids, mainly in human plasma, several HPLC methods which exploit spectrophotometric [21], spectrofluorometric [22][23][24], electrochemical [25,26], and mass spectrometry [27] detection have been developed and described in the literature. Although these assays allow quantification of LA in human plasma, they do not give the possibility to control levels of metabolically important endogenous amino thiols such as GSH, Cys, Hcy, and Cys-Gly.…”

Section: Reduced Formmentioning

confidence: 99%

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A Simplified Method for Simultaneous Determination of α-Lipoic Acid and Low-Molecular-Mass Thiols in Human Plasma

Borowczyk

Olejarz

Chwatko

et al. 2020

IJMS

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α-Lipoic acid, glutathione, cysteine, and cysteinylglycine can be applied as therapeutic agents in civilization diseases such as diabetes mellitus, cardiovascular diseases, and cancers. On the other hand, a higher concentration of homocysteine can result in health problems and has been indicated as an independent risk factor for cardiovascular disease and accelerated atherosclerosis. Here, the first simplified HPLC-UV assay that enables simultaneous determination of α-lipoic acid and low-molecular-mass thiols in plasma, reduces the number of steps, shortens the total time of sample preparation, and limits the amount of single-use polypropylene laboratory materials is described. The assay is based on reversed-phase high performance liquid chromatography with UV detection and simultaneous reduction of disulfide bound with tris(2-carboxyethyl)phosphine and the selective pre-column derivatization of the thiol group with 1-benzyl-2-chloropyridinium bromide. Linearity in the detector responses for plasma samples were observed in ranges: 0.12–5.0 nmol mL−1 for α-lipoic acid; 2.0–20.0 nmol mL−1 for glutathione, cysteinylglycine, and homocysteine; and 40.0–400.0 for cysteine. The LODs for α-lipoic acid and low-molecular-mass thiols were 0.08 and 0.12 nmol mL−1, respectively, while LOQs were 0.12 and 0.16 nmol mL−1, respectively. The usefulness of the proposed method has been proven by its application to real samples.

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Section: Resultsmentioning

confidence: 99%

Section: Reduced Formmentioning

confidence: 99%

A Simplified Method for Simultaneous Determination of α-Lipoic Acid and Low-Molecular-Mass Thiols in Human Plasma

Borowczyk

Olejarz

Chwatko

et al. 2020

IJMS

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“…In order to overcome this problem, derivatization reaction was utilized with the use of BCPB that is well known as the suitable reagent for thiols derivatization . It has been earlier proven that BCPB could also effectively react with LA , according to a general derivatization reaction scheme shown in Fig. .…”

Section: Resultsmentioning

confidence: 99%

“…Due to biological importance of LA, significant efforts have been made to develop a robust and reliable methods for its determination in biological fluids . Several methods were used for the detection and measurement of LA in urine or plasma including HPLC‐FL , HPLC‐UV , HPLC‐EC , HPLC‐MS , and MALDI‐TOF MS .…”

Section: Introductionmentioning

confidence: 99%

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Determination of lipoic acid in human urine by capillary zone electrophoresis

Kubalczyk

Głowacki

2017

Electrophoresis

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Fast, simple, and accurate CE method enabling determination of lipoic acid (LA) in human urine has been developed and validated. LA is a disulfide-containing natural compound absorbed from the organism's diet. Due to powerful antioxidant activity, LA has been used for prevention and treatment of various diseases and disorders, e.g. cardiovascular diseases, neurodegenerative disorders, and cancer. The proposed analytical procedure consists of liquid-liquid sample extraction, reduction of LA with tris(2-carboxyethyl)phosphine, derivatization with 1-benzyl-2-chloropyridinium bromide (BCPB) followed by field amplified sample injection stacking, capillary zone electrophoresis separation, and ultraviolet-absorbance detection of LA-BCPB derivative at 322 nm. Effective baseline electrophoretic separation was achieved within 6 min under the separation voltage of 20 kV (∼80 μA) using a standard fused-silica capillary (effective length 51.5 cm, 75 μm id) and BGE consisted of 0.05 mol/L borate buffer adjusted to pH 9. The experimentally determined limit of detection for LA in urine was 1.2 μmol/L. The calibration curve obtained for LA in urine showed linearity in the range 2.5-80 μmol/L, with R 0.9998. The relative standard deviation of the points of the calibration curve was lower than 10%. The analytical procedure was successfully applied to analysis of real urine samples from seven healthy volunteers who received single 100 mg dose of LA.

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Application of simultaneous separation and derivatization for the determination of α‐lipoic acid in urine samples by high‐performance liquid chromatography with spectrofluorimetric detection

Borowczyk

Olejarz

Chwatko

2019

Biomedical Chromatography

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To help to clarify therapeutic functions of lipoic acid (LA) in biochemical and clinical practice we have elaborated a fast, simple and accurate HPLC method enabling determination of LA in human urine. The proposed analytical approach includes reduction of LA with tris(2‐carboxyethyl)phosphine and simultaneous separation and derivatization of the analyte with butylamine and o‐phthaldialdehyde followed by spectrofluorimetric detection at λex = 340 nm and λem = 440 nm. The assay was performed using gradient elution and the mobile phase containing 0.0025 mol L−1 o‐phthaldialdehyde in 0.0025 mol L−1 NaOH and acetonitrile. Linearity of the detector response for LA was observed in the range of 0.3–8 μmol L−1. Limits of detection and quantification for LA in urine samples were 0.02 and 0.03 μmol L−1, respectively. The total analysis time, including sample work‐up, was <20 min. The analytical procedure was successfully applied to analysis of real urine samples delivered from six healthy volunteers who received a single 100 mg dose of LA.

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Determination of lipoic acid in human plasma by high-performance liquid chromatography with ultraviolet detection

Cited by 17 publications

References 28 publications

A Simplified Method for Simultaneous Determination of α-Lipoic Acid and Low-Molecular-Mass Thiols in Human Plasma

A Simplified Method for Simultaneous Determination of α-Lipoic Acid and Low-Molecular-Mass Thiols in Human Plasma

Determination of lipoic acid in human urine by capillary zone electrophoresis

Application of simultaneous separation and derivatization for the determination of α‐lipoic acid in urine samples by high‐performance liquid chromatography with spectrofluorimetric detection

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