Aleš Eisner

Comparison of pressurised fluid and ultrasonic extraction methods for analysis of plant antioxidants and their antioxidant capacity

The analytical method based on the high-performance liquid chromatography coupled with UV detection (HPLC/UV) for determination of selected antioxidants (i.e., esculetin, scopoletin, 7-hydroxycoumarine, rutin, xanthotoxin, 5-methoxypsoralen and quercetin) in plant material was developed. Pressurised fluid extraction (PFE) and ultrasonic extraction (USE) methods for the isolation of these compounds from ten real plant samples were used. Both extraction methods were optimised and compared to each other. For the proposed HPLC/UV method the LOQ values (limit of quantification) in the range from 22.7 (xanthotoxin) to 97.2 ng mL−1 (rutin) were obtained. For all extracts the antioxidant capacity based on the reduction of free 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) was also determined. Results ranged from 82.04 to 94.43% of DPPH radical inhibition for PFE method and from 76.01 to 89.94% in the case of USE method.

Extraction of antioxidants from plants using ultrasonic methods and their antioxidant capacity

The analytical method based on the HPLC coupled with UV detection (HPLC-UV) for the determination of selected antioxidants (i.e. esculetin, scopoletin, 7-hydroxycoumarine, rutin, xanthotoxin, 5-methoxypsoralen and quercetin) in plant material was developed. Two ultrasonic extraction methods for the isolation of these compounds from the plants such as Mentha longifolia L., Mentha spicata L., Ruta graveolens L., Achyllea millefolium L., Plantago lanceolata L. and Coriandrum sativum L. were used. Both of these methods, i.e. ultrasonic probe and ultrasonic bath, were optimised and compared to each other. For the proposed HPLC-UV method LOQ values in the range from 22.7 (xanthotoxin) up to 97.2 ng/mL (rutin) were obtained. For all extracts the antioxidant capacity based on the reduction of free 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical was also determined. Obtained results ranged from 10.11 up to 73.50% of DPPH radical inhibition.

Headspace single‐drop microextraction of herbal essential oils

A method employing the headspace single-drop microextraction (HS-SDME) is presented for the determination of essential oils in dried herbal leaves. By optimising the key experimental parameters, a linear response for the individual target compounds was obtained in the concentration range from LOQ to 4 mg/mL (r(2) = 0.9912-0.9998), with LODs from 3.3 up to 20.5 microg per 100 g of dried leaves, and the repeatability within the RSD of 2.1-8.9%. The HS-SDME-based procedure, enabling a rapid and simple analysis of essential oils in herbs, was applied to selected real samples (nine essential oils in four different samples) in combination with GC-FID identification and quantification of the target volatiles.

Comparison of supercritical fluid and Soxhlet extractions for the isolation of nitro compounds from soils

Supercritical fluid extraction (SFE) with CO(2), a clean and rapid alternative to conventional Soxhlet extraction, was investigated for the extraction of nitro compounds from soil samples. Quantitative extraction by SFE was accomplished at a pressure of 25 MPa and an extraction temperature of 60 degrees C, for 30 min in dynamic mode and using acetonitrile as modifier, and the results were comparable with those obtained by acetonitrile Soxhlet extraction (3 h) for all soil samples. Extracts from these two procedures were analyzed by gas chromatography coupled with mass spectrometry. Quantitative reproducibility for SFE extracts was acceptable (RSD 2-10%), and the quantity of solvent was reduced from 160 mL for Soxhlet extraction to 5 mL in the case of SFE.

Solid-phase microextraction based method for determination of essential oils components in herbal beverages

A method employing the direct immersion solid-phase microextraction followed by GC-MS analysis is presented for the determination of essential oils components in herbal tea infusions, i.e. their direct content in the liquid phase. The extraction performances were compared using five different microextraction fibres. Significant parameters affecting sorption process such as sample amount, sorption and desorption time and temperature, stirring speed, pH adjustment and effect of ionic strength were optimised and discussed. By optimising the key parameters, a detection limits (LOD = S/N × 3) for ten target marker compounds were obtained in the range from 5.3 to 48.2 ng/mL with recoveries ranged between 93.03 and 100.50%. Intra-day and inter-day repeatability at three concentration levels were found to be 1.1-15.3 and 7.2-15.5% RSD, respectively. Finally, the optimised procedure enabling a rapid and simple analysis of essential oils was applied for the direct determination of these compounds in ten herbal tea infusions.

Determination of 2-ethylhexyl nitrate in diesel fuel.

The aim of this study was to develop and validate fast and easily applicable GC/MS assay for the quantification of the substance that increases cetane number in diesel fuel (2-ethylhexylnitrate, 2-EHN). These requirements were fulfilled best by a headspace GC-MS assay with negative chemical ionization with methane (HS-GC/MS). Chromatographic separation is achieved using a DB5-MS capillary column after the addition of known amount of internal standard (o-nitrotoluene). The limit of detection was 0.009% v/v for 2-EHN and the limit of quantification was 0.03% v/v. The HS-GC/MS method was applied for the quantification of cetane improver in spiked diesel fuel and real diesel fuel. The method is linear over the studied range (0.03-0.3%, v/v), with satisfactory intra- and inter-assay precision, and the relative standard deviations are lower than 10%. Good accuracy is achieved with bias <10% at all levels tested.

Method validation for the determination of propellant components by Soxhlet extraction and gas chromatography/mass spectrometry.

A fast, sensitive, and accurate GC/MS method for the quantification of aliphatic nitroesters (ethylene glycol dinitrate, nitroglycerin, and triethylene glycol dinitrate) and aromatic amines (diphenylamine, 2-nitrodiphenylamine, and triphenylamine) in propellants was developed and validated. This method comprises a Soxhlet extraction step with dichloromethane, followed by separation on a capillary column MDN-5. Ionization of the analytes is carried out using electron ionization. The limit of quantification of the method was 1% w/w for aliphatic nitroesters and 0.1% w/w for aromatic amines (diphenylamine and triphenylamine). Values of repeatability and reproducibility for analyzed compounds were smaller than values of the maximum allowed tolerances of the Horwitz-equation RSD(max) and 2/3 RSD(max). Values of accuracy for selected compounds were below the acceptable threshold of 15% for all tested levels in the range of calibration curve excepting the lowest concentration of calibration curve for nitroglycerin and aromatic amines. During the validation of method, temperature instability in injection port of gas chromatograph and column was observed for 2-nitrodiphenylamine. Hence, it follows worse results of accuracy and linearity and 2-nitrodiphenylamine was not validated successfully.

Comparison of Two Ultrasonic Systems with Different Settings for Extraction of Capsaicinoids from Chili Peppers

Abstract Two ultrasonic systems were compared with different settings for extraction of capsaicinoids (capsaicin, dihydrocapsaicin and nordihydrocapsaicin) from chili peppers. The first system was ultrasonic homogenizer using an ultrasonic probe operating at a working frequency of 20 kHz, with variable setting of power output. The second one was ultrasonic cleaner operating at a working frequency of 35 kHz, with fixed power output of 50 W. Determination of the optimum extraction procedures was performed by comparison of the extraction yields. Content of capsaicinoids was determined by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Optimization studies of extractions by both ultrasonic systems were performed. Ten various real samples of chili underwent optimum extraction conditions in both ultrasonic systems. The results were compared with each other and to a conventional Soxhlet extraction. The ultrasonic methods provide the higher extraction efficiency and simple and straightforward operation. System working at higher power of ultrasound resulted in higher extraction yields. An application of ultrasonic system working at lower frequency of ultrasonic waves is slightly advantageous.