Triacylglycerol profiling by using chromatographic techniquesTriacylglycerols (TGs) make up the major part of naturally occurring fats and oils. The composition and fine structure of TGs determine to a large extent the functionality of fats and oils as food ingredients and the physiological effects of fats and oils as component of the human diet. Analysis of intact TGs is usually performed by chromatographic methods. In this article the application of gas-liquid chromatography, highperformance chromatography in normal and reversed phase mode, thin-layer chromatography and supercritical fluid chromatography for the qualitative and quantitative determination of TGs is reviewed. Emphasis is put on those factors that are decisive for obtaining reliable quantitative data. Furthermore, techniques for the stereospecific analysis of the fatty acid distribution along the glycerol backbone of TGs are presented briefly.
The value assignment for properties of six certified reference materials (ERM-AD623a–f), each containing a plasmid DNA solution ranging from 1 million to 10 copies per μL, by using digital PCR (dPCR) with the BioMark™ HD System (Fluidigm) has been verified by applying droplet digital PCR (ddPCR) using the QX100 system (Bio-Rad). One of the critical factors in the measurement of copy number concentrations by digital PCR is the partition volume. Therefore, we determined the average droplet volume by optical microscopy, revealing an average droplet volume that is 8 % smaller than the droplet volume used as the defined parameter in the QuantaSoft software version 1.3.2.0 (Bio-Rad) to calculate the copy number concentration. This observation explains why copy number concentrations estimated with ddPCR and using an average droplet volume predefined in the QuantaSoft software were systematically lower than those measured by dPCR, creating a significant bias between the values obtained by these two techniques. The difference was not significant anymore when the measured droplet volume of 0.834 nL was used to estimate copy number concentrations. A new version of QuantaSoft software (version 1.6.6.0320), which has since been released with Bio-Rad’s new QX200 systems and QX100 upgrades, uses a droplet volume of 0.85 nL as a defined parameter to calculate copy number concentration.Graphical AbstractMonolayer of droplets generated by the droplet generator and observed under an optical microscope
This is the ®rst study to report on the determination of Sb species in urine. To this end, HPLC was coupled online to an ICP-MS instrument using ultrasonic nebulization (USN) or hydride generation (HG) for sample introduction into the ICP-MS. The high chloride concentration in urine seriously hampered the chromatographic separation of Sb(V) and Sb(III) on the Dionex AS14 anion exchange column. Distinct signal suppression, shifting of retention times and severe peak broadening did not allow the application to urine samples. Progress to avoid these problems in HPLC-USN-ICP-MS could be made by employing a Hamilton PRP-X100 anion-exchange column. However, Na eluting in the void volume of the column gave rise to a Na-induced peak overlapping with the Sb(V) signal when USN was used to aspirate the HPLC eluents into the plasma. Therefore, a HG system was placed between the HPLC and ICP-MS instrumentation to overcome this dilemma. Thus, Sb(V) and Sb(III) were separated in urine with the PRP-X100 column using 20 mM EDTA at pH 4.7 as the mobile phase. Similarly, an ION-120 anion-exchange column was employed to separate trimethylantimony dichloride (TMSbCl 2) and Sb(V) with a mobile phase containing 2 mM NH 4 HCO 3 and 1 mM tartaric acid at pH 8.5. Detection limits of 20 ng l 21 , 12 ng l 21 and 8 ng l 21 for Sb(V), TMSbCl 2 and Sb(III), respectively, could be established in a 1z2 diluted urine matrix. The developed HPLC-HG-ICP-MS method was applied to the speciation of Sb in the urine of occupationally exposed and non-exposed subjects. Additionally, two lyophilised urine reference materials were investigated. Sb(V) was by far the predominant Sb species, followed by TMSbCl 2. Only ultratraces of Sb(III), if any detectable, could be found. The sum of the concentrations of Sb(V), Sb(III) and TMSbCl 2 in urine samples ranged between 51 and 78% of their total Sb concentrations.
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