Determination of total safranal by in situ acid hydrolysis in supercritical fluid media: Application to the quality control of commercial saffron

A procedure allowing hydrolysis reactions to be conducted in a dynamic supercritical-CO₂ medium was developed for quantifying total safranal (viz. free safranal present in the sample + safranal resulting from picrocrocin hydrolysis), which are the main component of the essential oil and responsible for the characteristic aroma of saffron. The proposed method allows total safranal amounts over the ranges 0.05-1.5 mg mL⁻¹ to be determined. The standard deviation achieved was 2%. This method was applied to the determination of safranal in natural saffron samples. The results obtained were compared with the “safranal value” total index, which is widely used as a quality measure of saffron products. The comparison revealed that the proposed method provides useful information not contained in the safranal value, based on the fact that, some samples with a high “safranal index” contain low concentrations of safranal. The proposed method is very useful for quality control in commercial saffron samples.     

Characterization of volatile components of Iranian saffron using factorial-based response surface modeling of ultrasonic extraction combined with gas chromatography–mass spectrometry analysis

The volatile components of Iranian saffron were extracted using ultrasonic solvent extraction (USE) technique and then were separated and detected by gas chromatography–mass spectrometry (GC–MS). Variables affecting the extraction procedure were screened by using a 2⁵⁻¹ fractional factorial design and among them; sample amount, solvent volume, solvent ratio and extraction time were optimized by applying a rotatable central composite design (CCD). The optimum values of factors were: 2.38 g sample, 29.04 mL solvent, 69.23% MeOH solvent ratio and 71.8 min for the extraction time. Forty constituents were identified for Iranian saffron by GC–MS representing 90% of the total peak area. The major components were 2,6,6-trimethyl-1,3-cyclohexadiene-1-carboxaldehyde, namely safranal (26.29%), bicyclo[3,2,0]hept-2-ene-,4-ethoxy-,endo (5.69%), linoleic acid (4.77%), 4-hydroxy-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde, namely HTCC (4.44%), and nonadecanol (3.32%). Some new compounds were identified for the first time in saffron. In addition, the results of this study were compared with those of Greek saffron.     

超临界流体技术在萃取中的应用简介

超临界流体萃取技术具有许多传统技术所没有的快速、高效、高选择性、低能耗等优点,特别是超临界流体的特性是它可以代替高毒有机溶剂做反应介质,符合绿色化学的要求。本文综述了超临界流体的特性,介绍了超临界流体萃取的原理,归纳了超临界流体萃取的应用现状,并介绍了超临界流体的其他领域应用概况。     

Determination of [13C-15N] Labeled Methylxanthines by Gas Chromatography-Atomic Emission Detection (GC-AED) and Compound-Independent Calibration

ABSTRACT

Compound independent calibration (CIC) can be used to measure stable isotopically labeled molecules and their metabolites. This study was designed to demonstrate that gas chromatography coupled to atomic emission detection and CIC are useful analytical tools for the specific and quantitative determination of ¹³C and ¹⁵N labeled caffeine and its metabolites. These labeled compounds can be measured by monitoring either ¹³C or ¹⁵N atomic emission lines and using only one calibration function from one of the analytes. This method can be helpful when it is difficult or impossible to obtain an authentic reference material, which is often the case in metabolic studies.     

Determination of Saffron Volatiles by HS-SBSE-GC in Flavored Cured Ham

At present, the development of new agri-food products, including flavored meat products presented in ready-to-eat vacuum packs, is encouraged. The addition of ingredients used as flavoring agents creates the need to be able to determine the volatile compounds responsible for their characteristic aroma. The aim of this study is to propose, develop, and validate a new method that uses headspace-stir bar sorptive extraction-gas chromatography/mass spectrometry (HS-SBSE-GC/MS) to determine the saffron aroma in cured ham flavored with this spice. Results showed that safranal was the main volatile compound that could be identified and quantified in cured ham flavored with saffron. This analytical method was adequate in terms of linearity, selectivity, sensitivity, and accuracy. To our knowledge, this is the first time that an HS-SBSE-GC/MS method for determining the saffron aroma of flavored cured ham has been developed and validated, and it is of interest to agri-food industries.     

Hydrogen and Atom Transfer Activity of Saffron Extracts by Square Wave Voltammetry

Saffron is an edible spice with highly appreciated sensory and antioxidant properties. One of the most representative redox species found in saffron extracts is crocin, whose content is used to evaluate the quality and value of the resulting spice. In this study, a voltammetry method based on the direct detection of crocin at a bare glassy carbon electrode is presented. The principle of the method is based on the monitoring of the anodic wave exhibited by crocin (0.1–1.0 mM) after its mixing with the azo radical initiator AAPH (20 mM) in ethanol:acetonitrile (1 : 1) solution. The decay rate of the anodic peak (E=+434 mV vs. Ag/Ag⁺), as a result of the consumption of crocin by AAPH, was used as index of the hydrogen transfer capacity and, thus, of its antioxidant activity. With a decay rate of k=0.02 h⁻¹, crocin exhibits only a weak antioxidant activity in comparison with tocopherols (k=0.13 h⁻¹), but still sufficient to protect against the oxidation of safranal, a further redox species found in saffron extracts and mainly responsible for its flavor. The proposed approach was finally applied to discriminate saffron extract samples from different geographical origins. The proposed approach is suitable to characterize the quality of saffron extracts and estimate its antioxidant properties.     

Evaluation of extraction/digestion techniques used to determine lead isotopic composition in forest soils

Lead isotopic studies in soils provide an efficient tool for tracing the sources of lead pollution. Five different extraction/digestion techniques (0.05 M EDTA, 0.5 M HNO₃, 2 M HNO₃, aqua regia, total digestion) were used for lead isotopic composition (²⁰⁶Pb/²⁰⁷Pb) determination in three forest soil profiles with different kinds of prevailing Pb contamination (unpolluted area, smelting area and vicinity of a motorway). The results obtained showed that all extraction/digestion methods used for the determination of ²⁰⁶Pb/²⁰⁷Pb ratios in surface horizons containing high organic matter contents gave statistically identical values (according to the Tukey test). In mineral soil horizons, differences between the individual extraction/digestion methods could be observed (the lowest ²⁰⁶Pb/²⁰⁷Pb ratios were obtained from EDTA extracts, corresponding to weakly bound anthropogenic lead, and the highest ²⁰⁶Pb/²⁰⁷Pb ratios were obtained from total digestion). The combination of total digestion and EDTA extraction (labile lead fraction) seems to be the optimal combination for ²⁰⁶Pb/²⁰⁷Pb ratio determination and optimal result interpretation.     

The Use of the PREPTM System for Handling of Body Fluids

Abstract

The Du Pont PREP^TM automated sample processor is a centrifugally based, microprocessor controlled instrument that was designed for extraction of samples from biological fluids. Extraction takes place in cartridges containing either organic resins or bonded silica packings as extraction sorbants. This paper will discuss the application of several lipophilic and ion exchange sorbants to the extraction of biological samples from body fluids. The advantages of these different types of sorbants will be compared and their performance with automated sample preparation will be shown. A variety of applications including the extraction of benzodiazepine, barbiturate, aminoglycoside and anticonvulsant drugs and their metabolites from serum, urine, and tissue homogenates will be discussed.     

CO LMR spectrum of14N16O and its analysis with spectra of14N17O and14N18O

LMR spectra for v=1←0 transitions of¹⁴N¹⁶O in X²II_{1/2, 3/2} states were observed at 5.6 μm and 5.4 μm of CO laser. Introducing the advanced isotopic molecular constant scaling function to Hund’s case (a) diatomic structure model, these spectra were analyzed and fitted together with all reliable previous spectral data of¹⁴N¹⁶O as well as¹⁴N¹⁷O and¹⁴N¹⁸O. A full set of precise molecular parameters and their vibrational dependencies have been determined with much higher precision (1–2 orders for most parameters). Many of them have been obtained for the first time. Using isotopic scaling function, the molecular constants of¹⁴N¹⁷O and¹⁴N¹⁸O were deduced.     

Gradient separation of pharmaceuticals employing supercritical CO2 and modifiers

Sub- and Supercritical Fluid Chromatography (SFC) were used to separate seven purine and pyrimidine-based drugs. A two-pump system was used, where one pump delivered the carbon dioxide, and the other pump delivered modifier. The modifiers used in this work were methanol, and methanol containing 0.001 M tetrabutylammonium hydroxide. Two of the compounds studied were hydrochloride salts. These compounds were analyzed by a Supercritical Fluid Extraction (SFE)/SFC method.     

Extraction and Spectrophotometric Determination of Manganese(VII) with N1-Hydroxy-N1,N2-Diphenylbenzamidine

Abstract

Spectrophotometric determination of manganese(VII) at 614 nm after its extraction with N¹ -hydroxy-N¹,N²-diphenylbenzamidine into amyl alcohol at pH 7–8 is described. Beer's law was obeyed for 0.1–10 μg ml^?1 Mn(VII). The effects of experimental variables and of several diverse ions on the determination of manganese(VII) have been studied. The method has been applied to the determination of manganese in steels and in water extracts of a commercial tea and is found to be simple, precise and highly selective.     

Use of supercritical fluid extraction in the analysis of polychlorinated dibenzodioxins and dibenzofurans

Summary Supercritical fluid extraction is a powerful technique with great promise in organic analytical chemistry. To date little has been published on the use of SFE in the analysis of polychlorinated dibenzodioxins and dibenzofurans (PCDD/F). The data point, however, to the feasibility of a selective and exhaustive extraction of these compounds. Solid phase trapping of the extracted PCDD/F allows for on-line class separation and clean-up and seems to be the most flexible choice of collection mode. Both CO₂ and N₂O can be used as supercritical fluids. Extraction recoveries can be improved with a small percentage of an organic solvent added to the supercritical fluid as modifier. Methanol and benzene have proven to be efficient. Relatively strong supercritical fluid conditions are needed for the extraction of PCDD/F from fly ash (350–400 atm at 330–370 K). Spiked internal standards are easily extracted even at mild conditions whereas native PCDD/F are not; thus caution should be taken when an isotopic dilution technique is used for a future evaluation of SFE in the analysis of PCDD/F.     

Effect of ultrasonic waves on crocin and safranal content and expression of their controlling genes in suspension culture of saffron (Crocus sativus L.)

The expression of biosynthesis controlling genes of crocin and safranal in saffron (Crocus sativus) can be influenced by ultrasonic waves. Sterilized saffron corms were cultured in a ½-MS medium supplemented by 2-4-D and BAP. Saffron callus cells were treated with ultrasonic waves in a cellular suspension culture under optimal growth conditions. The samples were collected at 24 and 72 hours after treatment in three replications. The secondary metabolites were measured by high-performance liquid chromatography and the gene expression was analysed by the real-time polymerase chain reaction. Results indicate that this elicitor can influence the expressions of genes CsBCH, CsLYC and CsGT-2; the ultrasonic waves acted as an effective mechanical stimulus to the suspension cultures. The analysis of variance of the ultrasonically produced amounts of safranal and crocin indicates that there is a significant difference between once- and twice-treated samples in that the amount of safranal was the highest within the samples taken from the twice-treated suspension culture at 72 h after the ultrasound treatment, and the crocin was maximised after 24 h passed the twice-applied ultrasound treatment.     

Volatile Analysis of Wuliangye Baijiu by LiChrolut EN SPE Fractionation Coupled with Comprehensive GC×GC-TOFMS

Wuliangye baijiu is one of the most famous Chinese liquors with a protected geographical indication. This study used LiChrolut^® EN-based solid-phase extraction (SPE) and fractionation combined with comprehensive two-dimensional chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS) to unveil its volatile composition. The volatiles were isolated with LiChrolut^® EN-based SPE and traditional liquid-liquid extraction (LLE). The neutral/basic fractions from LLE and the SPE were fractionated on a LiChrolut^® EN SPE column and analyzed by comprehensive GC×GC-TOFMS. Compared with LLE, more esters and alcohols were detected in the SPE-based extraction. The SPE fractionation and GC×GC-TOFMS analysis resulted in the identification of about 500 volatile compounds in more than 3000 peaks of the Wuliangye baijiu. The approach simplifies the complex baijiu composition into functional group-based fractions for reliable identification and analysis. This study provided a confidence volatile identification approach for Chinese baijiu based on the SPE fractionation GC×GC-TOFMS.     

Use of volatile organic solvents in headspace liquid‐phase microextraction by direct cooling of the organic drop using a simple cooling capsule

A low‐cost and simple cooling‐assisted headspace liquid‐phase microextraction device for the extraction and determination of 2,6,6‐trimethyl‐1,3 cyclohexadiene‐1‐carboxaldehyde (safranal) in Saffron samples, using volatile organic solvents, was fabricated and evaluated. The main part of the cooling‐assisted headspace liquid‐phase microextraction system was a cooling capsule, with a Teflon microcup to hold the extracting organic solvent, which is able to directly cool down the extraction phase while the sample matrix is simultaneously heated. Different experimental factors such as type of organic extraction solvent, sample temperature, extraction solvent temperature, and extraction time were optimized. The optimal conditions were obtained as: extraction solvent, methanol (10 μL); extraction temperature, 60°C; extraction solvent temperature, 0°C; and extraction time, 20 min. Good linearity of the calibration curve (R² = 0.995) was obtained in the concentration range of 0.01–50.0 μg/mL. The limit of detection was 0.001 μg/mL. The relative standard deviation for 1.0 μg/mL of safranal was 10.7% (n = 6). The proposed cooling‐assisted headspace liquid‐phase microextraction device was coupled (off‐line) to high‐performance liquid chromatography and used for the determination of safranal in Saffron samples. Reasonable agreement was observed between the results of the cooling‐assisted headspace liquid‐phase microextraction high‐performance liquid chromatography method and those obtained by a validated ultrasound‐assisted solvent extraction procedure.     

Isotopic Traceability (13C and 18O) of Greek Olive Oil

In recent years, isotopic analysis has been proven a valuable tool for the determination of the origin of various materials. In this article, we studied the ¹⁸O and ¹³C isotopic values of 210 olive oil samples that were originated from different regions in Greece in order to verify how these values are affected by the climate regime. We observed that the δ¹⁸O isotopic values range from 19.2 ‰ to 25.2 ‰ and the δ¹³C values range from −32.7 ‰ to −28.3 ‰. These differences between the olive oils’ isotopic values depended on the regional temperature, the meteoric water, and the distance from the sea. Furthermore, we studied the ¹³C isotopic values of biophenolic extracts, and we observed that they have same capability to differentiate the geographic origin. Finally, we compared the isotopic values of Greek olive oils with samples from Italy, and we concluded that there is a great dependence of oxygen isotopes on the climatic characteristics of the different geographical areas.     

Volatilization of 14C-Labelled Fenpropimorph after Application to Plants and Soil under Simulated Outdoor Conditions

Abstract

The volatility of fenpropimorph was investigated in a laboratory chamber constructed for studying the volatilization of “4C-labelled pesticides from plant and soil surfaces. Summer barley was cultivated on experimental platforms (0.5 m²) filled with soil and treated in an application chamber with ¹⁴C-labelled fenpropimorph formulation (Corbel^R) at the beginning of ear emergence. After application, the platform was transferred into the volatilization chamber where a 96 h outdoor weather scenario was simulated. The results of three experiments demonstrated that up to 60% of the initial total radioactivity could be released from the plant-soil system within 96 h, most of it being the unchanged ¹⁴C-fenpropimorph which undergoes a fast oxidative (degradation by solar irridation in the atmoshpere. Furthermore, ¹⁴CO, was detected in quantities of 1.1 to 1.8%. After plant extraction, however, mainly polar metabolites, such as fenpropimorh acid, were found four days alter application by Radio-HPLC-analysis. In order to evaluate the volatilization behaviour of fenpropimorph sprayed to bare soils, three additional experiments were carried out showing a volatilization rate of 11.4% at most, much lower than those of plant surfaces.     

Isotopic finger-printing of active pharmaceutical ingredients by 13C NMR and polarization transfer techniques as a tool to fight against counterfeiting

The robustness of adiabatic polarization transfer methods has been evaluated for determining the carbon isotopic finger-printing of active pharmaceutical ingredients. The short time stabilities of the adiabatic DEPT and INEPT sequences are very close to that observed with the one pulse sequence, but the DEPT long time stability is not sufficient for isotopic measurements at natural abundance or low enrichment. Using the INEPT sequence for ¹³C isotopic measurements induces a dramatic reduction in the experimental time without deterioration in short time or long time stability. It appears, therefore, to be a method of choice for obtaining the isotopic finger-print of different ibuprofen samples in a minimum time. The results obtained on 13 commercial ibuprofen samples from different origins show that this strategy can be used effectively to determine ¹³C distribution within a given molecule and to compare accurately differences in the isotopic distribution between different samples of the given molecule. The present methodology is proposed as a suitable tool to fight against counterfeiting.     

Setting up of an Isotope Analyser for Quantitative Determination of Percentage Abundance of 15N in Nitrogen Gas

Abstract

An ¹⁵N isotope analyser has been assembled for the quantitative determination of the percentage abundance of ¹⁵N isotope in nitrogen gas employing molecular optical emission spectroscopic technique. The (2,0) band of the second positive system of N₂ was excited in a high frequency discharge. The band heads of the isotopic species ¹⁴N¹⁴N and ¹⁴N¹⁵N molecules were resolved using a 0.5 m monochromator and their intensity peaks were measured. The ratio of the peak heights enabled the quantitative determination of ¹⁵N in the N₂ sample. The performance of the isotope analyser assembled was evaluated and it was found to be quite good as inferred from the analysis of primary standards. The analytical error in the ¹⁵N concentration-range 0.36 to 24% is <6% and rsd is <4%. A salient feature of the method adopted is that it is direct and does not call for the use of comparision standards.