A new method to simultaneously quantify the antioxidants: carotenes, xanthophylls, and vitamin A in human plasma

A simple and accurate reversed phase high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) method for simultaneously determining and quantifying the antioxidants carotenes, xanthophylls, and retinol in human plasma is presented in this paper. Compounds were extracted with hexane, a C30 column, and a mobile phase of methanol, methyl tert-butyl ether, and water were used for the separation of the compounds. A total of 8 carotenoids, 3 Z--carotene isomers, and 1 fat-soluble vitamin (retinol) were resolved within 72 min at a flow rate of 0.6 mL/min. Detection was achieved at 450 nm for carotenoids and 330 nm for retinol. To evaluate the effectiveness of themethod, it has been applied to an intervention study conducted on eight volunteers. Results. Limits of detection were between 0.1 g/mL for lycopene and astaxanthin and 1.3 g/mL for 15-Z--carotene. Recoveries were ranged between 89% and 113% for -carotene and astaxanthin, respectively. Accuracy was between 90.7% and 112.2% and precision was between 1% and 15% RSD. In human plasma samples compounds studied were identified besides three lycopene isomers, demonstrated to be suitable for application in dietary intervention studies. Conclusions. Due to its accuracy, precision, selectivity, and reproducibility, this method is suitable to dietary habits and/or antioxidants status studies

Efecto del licopeno y los carotenoides del tomate en marcadores inflamatorios de la aterosclerosis en pacientes de riesgo cardiovascular

[spa] El tomate es una de las hortalizas más consumidas en todo el mundo y tanto su demanda como su producción han aumentado progresivamente en los últimos 20 años. Europa se encuentra entre los principales productores mundiales de este fruto, siendo España uno de los diez países con mayor cosecha de tomates tanto para la industria, como para el consumo en fresco. Además de sus propiedades sensoriales y de su versatilidad culinaria, que lo convierten en un alimento apto para consumo en crudo, así como en diferentes preparaciones, el tomate es considerado una fuente importante de nutrientes, debido a su elevado contenido de compuestos bioactivos, entre los que destacan los carotenoides. Los carotenoides son una familia de compuestos de más de 600 pigmentos vegetales liposolubles que se encuentran tanto en organismos fotosintéticos, como no fotosintéticos y son responsables de las coloraciones amarillas, anaranjadas y rojas que se observan en la naturaleza. El licopeno es uno de los carotenoides más abundantes del tomate, el cual es responsable de su coloración roja. En los alimentos crudos, el licopeno se encuentra principalmente en la configuración trans, mientras que en el plasma y en los tejidos, la configuración predominante es la cis. El estudio de los carotenoides con relación a la salud, y muy especialmente del licopeno, ha cobrado mucha relevancia en los últimos años, ya que se ha visto que su consumo regular se relaciona con un menor riesgo de desarrollar enfermedades crónico-degenerativas, tales como las enfermedades neurodegenerativas, distintas formas de cáncer, así como de enfermedades cardiovasculares. Entre las enfermedades cardiovasculares, la aterosclerosis es una de las responsables de más muertes y comorbilidades en todo el mundo y tanto en su inicio como en su progresión, se encuentran implicados múltiples procesos inmunoinflamatorios. La determinación de los efectos de los carotenoides del tomate sobre la expresión de moléculas inflamatorias y quimiotácticas implicadas en el desarrollo y la progresión de la aterosclerosis, así como el esclarecimiento del papel biológico que cumplen los distintos isómeros del licopeno en la modulación de estas moléculas es de suma importancia para la implementación de estrategias dietéticas y nutricionales que contribuyan con la prevención del desarrollo de estas enfermedades

Development of an advanced HPLC-MS/MS method for the determination of carotenoids and fat-soluble vitamins in human plasma

The concentration of carotenoids and fat-soluble vitamins in human plasma may play a significant role in numerous chronic diseases such as age-related macular degeneration and some types of cancer. Although these compounds are of utmost interest for human health, methods for their simultaneous determination are scarce. A new high pressure liquid chromatography (HPLC)-tandem mass spectrometry (MS/MS) method for the quantification of selected carotenoids and fat-soluble vitamins in human plasma was developed, validated, and then applied in a pilot dietary intervention study with healthy volunteers. In 50 min, 16 analytes were separated with an excellent resolution and suitable MS signal intensity. The proposed HPLC-MS/MS method led to improvements in the limits of detection (LOD) and quantification (LOQ) for all analyzed compounds compared to the most often used HPLC-DAD methods, in some cases being more than 100-fold lower. LOD values were between 0.001 and 0.422 µg/mL and LOQ values ranged from 0.003 to 1.406 µg/mL, according to the analyte. The accuracy, precision, and stability met with the acceptance criteria of the AOAC (Association of Official Analytical Chemists) International. According to these results, the described HPLC-MS/MS method is adequately sensitive, repeatable and suitable for the large-scale analysis of compounds in biological fluids

Development of an Advanced HPLC–MS/MS Method for the Determination of Carotenoids and Fat-Soluble Vitamins in Human Plasma

The concentration of carotenoids and fat-soluble vitamins in human plasma may play a significant role in numerous chronic diseases such as age-related macular degeneration and some types of cancer. Although these compounds are of utmost interest for human health, methods for their simultaneous determination are scarce. A new high pressure liquid chromatography (HPLC)-tandem mass spectrometry (MS/MS) method for the quantification of selected carotenoids and fat-soluble vitamins in human plasma was developed, validated, and then applied in a pilot dietary intervention study with healthy volunteers. In 50 min, 16 analytes were separated with an excellent resolution and suitable MS signal intensity. The proposed HPLC–MS/MS method led to improvements in the limits of detection (LOD) and quantification (LOQ) for all analyzed compounds compared to the most often used HPLC–DAD methods, in some cases being more than 100-fold lower. LOD values were between 0.001 and 0.422 µg/mL and LOQ values ranged from 0.003 to 1.406 µg/mL, according to the analyte. The accuracy, precision, and stability met with the acceptance criteria of the AOAC (Association of Official Analytical Chemists) International. According to these results, the described HPLC-MS/MS method is adequately sensitive, repeatable and suitable for the large-scale analysis of compounds in biological fluids

Characterisation of bioactive compounds and assessment of antioxidant activity of different traditional Lycopersicum esculentum L. varieties: chemometric analysis

Eight different Serbian genotypes were analysed for their polyphenol, carotenoid, vitamin C content and evaluated for their antioxidant properties. The highest content of biologically important carotenoids such as lutein (4.58 mg/10 g), lycopene (160.64 mg/10 g) and beta-carotene (189.64 mg/10 g) were detected in the genotype S606. Rutin was the most abundant phenolic compound in all tastes samples, but its content is highest in the genotype S615 (1424.30 mu g/100 g dw). All tomato samples were the great source of vitamin C, where the sample S615 stood out (68.54 mg AA g(-1) of dw). Their content of antioxidant compounds suggested that genotypes S606 and S615 showed the best antioxidant potential. Principal component analysis (PCA) and Partial least squares (PLS) were applied to analyse results. The results obtained in the present study could be of considerable interest for breeding programmes wishing to select tomato genotypes with high biological and nutritional properties