Determination of taurine in biological samples and isolated hepatocytes by high-performance liquid chromatography with fluorimetric detection

doi:10.1016/0378-4347(94)80067-7

Journal of Chromatography B: Biomedical Sciences and Applications

Volume 657, Issue 1, 1 July 1994, Pages 37-45

https://doi.org/10.1016/0378-4347(94)80067-7 Get rights and content

Abstract

A high-performance liquid chromatographic method with fluorimetric detection is described for the routine and selective determination of taurine in urine, serum, tissues and isolated hepatocytes. The preparation and use of ion-exchange resins to extract taurine from biological samples is included. Taurine was derivatised with o-phthalaldehyde/2-mercaptoethanol prior to injection onto a C₁₈ column (LiChrospher^R 100 RP-18, 5 μm, 125 × 4 mm I.D.). Isocratic elution of the adduct was carried out using NaH₂PO₄ (0.05 M, pH 5.4) in methanol and water (43:57, v/v). Homoserine was used as an internal standard to facilitate the standardisation and quantitation of samples and analysis was completed in 6 min with homoserine and taurine eluting after 3 and 4 min, respectively. The method will detect 0.5 pmol of taurine on the column. Appropriate dilutions of these biological samples enable these samples to be assayed on an autosampler, using the same standard curve. Concentrations of taurine in human, dog and rat urine, rat liver, serum and isolated hepatocytes are reported.

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Reduced de novo synthesis of 5-methyltetrahydrofolate and reduced taurine levels in ethanol-treated chick brains
2010, Comparative Biochemistry and Physiology - C Toxicology and Pharmacology
In previous studies, exogenous ethanol (3 mmol EtOH/kg egg) caused a 1.6-fold increase in chick brain homocysteine (HoCys) levels at 11 days of development and the mixture of 3 mmol EtOH/kg egg and 34 μmol folic acid/kg egg attenuated EtOH-induced increases in chick brain HoCys levels. Because HoCys is converted to methionine utilizing the methyl donor, 5-methyltetrahydrofolate (5-methyl THF), we studied whether exogenous ethanol (3 mmol EtOH/kg egg) or the mixture of 3 mmol EtOH/kg egg and 34 μmol 5-methyl THF/kg egg inhibited chick brain 10-formyltetrahydrofolate dehydrogenase (10-FTHF DH; EC 1.5.1.6) activities and brain N⁵, N¹⁰-methylenetetrahydrofolate reductase (MTHFR; EC 1.5.1.20) activities at 11 days of development. Three daily dosages of 3 mmol EtOH/kg egg (E_0–2) caused approximately a 7-fold reduction in brain 10-FTHF DH activities and approximately a 1.9-fold reduction in brain MTHFR activities as compared to controls at 11 days of development (p ≤ 0.05). Because HoCys is also removed by the transsulfuration pathway, which synthesizes taurine, we studied whether exogenous ethanol (3 mmol EtOH/kg egg) or the mixture of 3 mmol EtOH/kg egg and 34 μmol 5-methyl THF/kg egg influenced chick brain taurine levels. In EtOH-treated and EtOH and 5-methyl THF-treated embryos, brain taurine levels decreased by approximately 5.5-fold and 6.2-fold as compared to controls, respectively (p ≤ 0.05). Exogenous 5-methyl THF failed to attenuate EtOH-induced decreased brain taurine levels at 11 days of development.
Taurine-deficient diet up-regulated cystathionine β-synthase monoallele in hemizygous cystathionine β-synthase knockout mice
2009, Nutrition Research
Citation Excerpt :
For liver supernatant taurine analysis, a Waters LC Module-1 Plus (comprised a Model 600 ternary solvent delivery system and a Model 715 autosampler) was used in conjunction with a Waters 474 fluorometer (Waters Corporation) at a flow rate of 2 mL/min, with the column temperature at 50°C at an excitation wavelength of 230 nm and emission wavelength of 389 nm. Waters Millennium-32 Chromotography Software was used to control the chromatographic integration, quantitation, and mobile phase [10]. Liver taurine concentrations were expressed as micromoles per gram of protein.
Impaired cystathionine β-synthase (CBS) causes hyperhomocystinuria and hyperhomocysteinemia, both risk factors for cardiovascular diseases. Reduced CBS activity could decrease cysteine and taurine biosyntheses (metabolites of homocysteine degradation) and lead to less taurocholic acid production with a resultant increased cholesterol content. We hypothesized that a deficiency in CBS genetic material and enzyme activity would reduce taurine synthesis, which would lead to an elevated cholesterol concentration. Both sexes of hemizygous C57BL/6J-Cbs^tm1Unc [CBS (+/−)] and wild-type C57BL/6J mice [CBS (+/+)] were divided into 2 groups. One group of CBS (+/−) and CBS (+/+) mice was fed a cysteine- and taurine-deficient diet for 8 weeks, and the other group was fed a cysteine, taurine, and vitamin B6–deficient diet for 8 weeks. Significantly higher plasma total homocysteine concentrations occurred in the CBS (+/−) mice than their CBS (+/+) cohorts. Female mice of both genotypes had significantly higher plasma total homocysteine concentrations and significantly lower relative CBS mRNA levels than did male mice. During vitamin B₆ deficiency, plasma total homocysteine concentrations were significantly elevated. Three important findings were a differential sex response of CBS mRNA to feeding the vitamin B₆ diet; CBS (+/−) mice had a significantly lower plasma cholesterol concentration, contrary to what was anticipated; and during feeding, the taurine- and cysteine-deficient diet, CBS mRNA levels in CBS (+/−) mice were reduced only 13% rather than the expected 50%. We conclude that the remaining CBS monoallele is up-regulated in mice when fed a taurine-deficient diet to produce additional CBS mRNA.
Exogenous folate ameliorates ethanol-induced brain hyperhomocysteinemia and exogenous ethanol reduces taurine levels in chick embryos
2009, Comparative Biochemistry and Physiology - C Toxicology and Pharmacology
The effects of exogenous ethanol and/or folic acid on endogenous homocysteine (HoCys) and SAM (S-adenosylmethionine)/SAH (S-adenosylhomocysteine) levels in chick brains were studied at 11 days of development. Embryonic EtOH (3.0 mmol/kg egg) exposure caused a 1.6-fold increase in brain HoCys levels and a 9-fold decrease in brain SAM/SAH levels as compared to controls (p ≤ 0.05). Brain HoCys and SAM/SAH levels returned to control values when injected with a mixture of EtOH and folic acid (3.0 mmol EtOH/kg egg and 34 μmol folic acid/kg egg). The effects of exogenous EtOH on the remethylation pathway, as measured by 10-formyltetrahydrofolate dehydrogenase (10-FTHF DH) activities, and the transsulfuration pathway, as measured by taurine levels, were studied at 18 days of development. A single dosage of EtOH (3.0 mmol/kg egg; E₀) and two daily dosages of EtOH (E_0–1) failed to influence brain and hepatic 10-FTHF DH activities when compared to controls. However, three daily dosages of EtOH (E_0–2) caused approximately a two-fold increase in brain 10-FTHF DH activities and a three-fold increase in hepatic 10-FTHF DH activities as compared to controls (p ≤ 0.05). Three daily EtOH dosages (E_0–2) caused reduced taurine levels in both brain and hepatic tissues (p ≤ 0.05). Meanwhile, a single EtOH dosage (E₀), two daily EtOH dosages (E_0–1), and three daily EtOH dosages (E_0–2), caused reduced hepatic taurine levels as compared to controls (p ≤ 0.05).
Site-specific sampling of taurine from rat brain followed by on-line sample pre-concentration, throughout in-capillary derivatization and capillary electrophoresis
2006, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
A method of pinpoint-sampling followed by on-line pre-concentration of the sample, throughout in-capillary derivatization and capillary electrophoretic separation was evaluated by demonstrating the detection of taurine, 2-aminoethanesulfonic acid at a specific location of a rat brain. The direct sampling of taurine from the rat brain was accomplished by using voltage injection associated with two kinds of driving forces, electrophoretic flow and electroosmotic flow (EOF). The capillary tube (75 μm of inner diameter × 375 μm of outer diameter) of the capillary electrophoresis (CE) apparatus was already filled with a CE run buffer, viz., 40 mM phosphate–borate buffer (pH 10) containing 2 mM o-phthalaldehyde (OPA)/N-acetylcysteine (NAC) as the derivatization reagent. One end of a platinum wire (0.5 mm o.d.), used as the anode, and the inlet end of capillary tube (from which a 1.0 cm long polyimide coating was removed), were pricked down onto the surface of either the cerebrum or cerebellum of a rat brain at a location of very small dimension. When a low voltage (5 kV, 30 s) was applied, taurine began to move from the rat brain into the capillary tube, and, simultaneously, electric focusing of taurine occurred by the action of “the pH-junction effect” at the inlet end of the capillary tube. After completing the injection, both the platinum wire and capillary tube were detached from the brain and dipped into the run buffer in an anode reservoir filed with the same solution as that in the capillary tube for the CE apparatus. Then, by applying a high voltage (20 kV) between the ends of the capillary tube, taurine was automatically derivatized to yield the fluorescent derivative, separated and detected with fluorescence (E_x = 340 nm, E_m = 455 nm) during migration throughout the capillary tube. The migration profiles obtained from cerebrum and cerebellum appeared to be different, but the peak corresponding to taurine was identified on both electropherograms. The efficacy of the present method including sample on-line pre-concentration prior to throughout in-capillary derivatization CE was first verified with several preliminary experiments by using samples of taurine in water, saline and a piece of 1.5% agar–gel block, as an alternate standard for the rat brain used in this study.
Development and validation of a high-performance liquid chromatography-mass spectrometry assay for methylxanthines and taurine in dietary supplements
2005, Journal of Pharmaceutical and Biomedical Analysis
A procedure based on liquid chromatography–mass spectrometry (LC–MS) is described for determination of caffeine, theobromine, theophylline, taurine in different dietary supplements. After addition of tryptophan as internal standard, both solid and liquid specimens were extracted with 4 ml of hexane/isopropanol (9:1).
Chromatography was performed on a C₁₈ reversed-phase column using water/methanol/acetic acid (75:20:5, v/v/v) as a mobile phase. Analytes were determined in LC–MS single ion monitoring mode with atmospheric pressure ionization-electrospray (ESI) interface. The method was validated in the range 0.1–500 and 0.06–500 μg/ml or μg/g for taurine and caffeine, respectively; 0.06–100 μg/ml or μg/g for theobromine and theophylline. Mean recoveries ranged between 70.1 and 94.4% for different analytes. The quantification limits were 0.1 μg/ml or μg/g for taurine and 0.06 μg/ml or μg/g for methylxanthines either in liquid samples or in solid samples. The method was applied to the analysis of various dietary supplements containing methylxanthines and taurine. Energetic drinks contained amounts of taurine in the range of hundreds to thousands μg/ml and ten times lower amounts of caffeine. Conversely, herbal powders, tablets and capsules mainly contained mg amounts of caffeine per gram of product with the other two methylxanthines in the range of ten to hundred μg/g.
Quantitative determination of taurine in real samples by high-performance anion-exchange chromatography with integrated pulsed amperometric detection
2004, Talanta
As taurine is a very important compound involved in a large number of metabolic processes, it is naturally present in the mammal tissues and is often deliberately added in some foods as a fortifying component. A detailed knowledge of taurine metabolic roles in biological systems can be obtained only if a sensitive, reliable and rapid analytical method is available. This article describes the successful application of high-performance anion-exchange chromatography coupled with integrated pulsed amperometric detection (HPAEC-IPAD) for taurine determination in egg white and yolk samples, as well extracts of human serum and urine. Applications are shown for determination of taurine in soft drinks and pharmaceutical preparations where the taurine content was evaluated by standard additions. These results were achieved without prior derivatization of taurine.

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Determination of taurine in biological samples and isolated hepatocytes by high-performance liquid chromatography with fluorimetric detection

Abstract

J. Chromatogr.

Anal. Biochem.

J. Chromatogr.

J. Chromatogr.

J. Chromatogr.

Anal. Biochem.

Anal. Biochem.

Clin. Chim. Acta

Arch. Biochem. Biophys.

Clin. Chim. Acta

Methods Enzymol.

J. Chromatogr.

Toxicology

Biochem. Pharm.

Toxicology

Ann. Rev. Biochem.

Physiol. Rev.