Determination of fat-soluble vitamins A, D2, D3, E and K3 by isotope dilution and LC-MS/MS instrument assembly

The purpose of our publication is the determination of the total amount (of natural origin and added) of fat-soluble vitamins A, D2, D3 and E in low amounts in foods (wheat flour, soft drinks, effervescent tablets) and dietary supplements using a liquid chromatography tandem mass spectrometry (LC-MS/MS) method. The samples were diluted with isotope-labeled derivatives of the target components (vitamin A-d6, vitamin D2-d3, vitamin D3-d3, vitamin E-d6), and after extraction and saponification, they were purified by liquid-liquid extraction. After a solvent exchange, the concentration of the vitamins was determined on a C8 HPLC column using acidic mobile phases (0.1% formic acid in water/methanol) and LC-MS/MS technique. In dietary supplements, the analysis of the fat-soluble vitamin K3 may also be important, because the use of vitamin K3 is currently not approved in human formulations. During the determination of vitamin K3, saponification is not necessary, due to its structure, alkaline hydrolysis would lead to the decomposition of vitamin K3, so this component was analyzed by a method different from the one used for the other vitamins. LC-MS/MS analysis of small amounts of vitamin K3 is more complicated than that of other vitamins due to the low sensitivity of the MS instrument to vitamin K3. The determination of vitamin K3 was therefore carried out after chemical derivatization with L-cysteine as a derivatizing reagent, also with isotope dilution and LC-MS/MS technique. After intralaboratory validation, the methods were successfully used in domestic and international proficiency tests in infant formulas and liquid vitamin preparations

Zsírban oldódó A-, D2-, D3-, E- és K3-vitaminok meghatározása izotóphígítással és LC-MS/MS műszeregyüttessel

Közleményünk célja élelmiszerekben (búzaliszt, üdítő, pezsgőtabletta) és étrendkiegészítőkben alacsony mennyiségben előforduló zsírban oldódó A-, D2-, D3-, E-vitaminok teljes (természetes eredetű és hozzáadott) meghatározása folyadékkromatográfiás tandem tömegspektrometriai (LC-MS/MS) módszerrel. A mintákat a célkomponensek izotópjelölt származékaival hígítottuk (A-vitamin-d6, D2-vitamin-d3, D3-vitamin-d3, E-vitamin-d6), majd az extrakciót és szappanosítást követően folyadék-folyadék extrakcióval tisztítottuk azokat. Egy oldószercserét követően C8-as HPLC oszlopon savas mozgófázisok (0,1% hangyasav vízben/ metanolban) alkalmazásával és LC-MS/MS technikával határoztuk meg a vitaminok koncentrációját. Táplálék kiegészítőkben fontos lehet a zsírban oldódó K3-vitamin vizsgálata is, mert a K3-vitamin alkalmazása humán készítményekben jelenleg nem engedélyezett. A K3-vitamin meghatározása során szappanosításra nincs szükség, szerkezetéből adódóan a lúgos hidrolízis a K3-vitamin bomlásához vezetne, így ezt a komponenst egy, a többi vitamintól eltérő módszerrel vizsgáltuk. A K3-vitamin kis mennyiségben történő LC-MS/MS jellegű vizsgálata a K3-vitamin MS készülékben mutatott alacsony érzékenysége miatt bonyolultabb, mint a többi vitaminé. A K3- vitamin meghatározását ezért L-ciszteinnel, mint származékképző reagenssel történő kémiai származékképzést követően végeztük, szintén izotóphígítással és LC-MS/MS technikával. A módszereket laboratóriumon belüli validálását követően hazai és nemzetközi körvizsgálatokban sikeresen alkalmaztuk csecsemő tápszerben és folyékony vitamin készítményben

A gyermekkori koronavírus-fertőzést követő sokszervi gyulladás diagnosztikája és kezelése

A SARS-CoV-2-fertőzés ritka gyermekkori szövődménye a sokszervi gyulladás, angol terminológiával paediatric inflammatory multisystem syndrome (PIMS). Két vagy több szerv érintettségével járó, súlyos tünetekkel induló betegségről van szó, amelynek tünetei átfedést mutatnak a Kawasaki-betegséggel, a toxikus sokk szindrómával és a makrofágaktivációs szindrómával. A PIMS-betegek intenzív terápiás osztályon vagy intenzív terápiás háttérrel rendelkező intézményben kezelendők, ahol biztosítottak a kardiológiai ellátás feltételei is. A szükséges immunterápia a klinikai prezentációtól függ. A jelen közleményben a szerzők a releváns nemzetközi irodalom áttekintését követően ajánlást tesznek a PIMS diagnosztikai és terápiás algoritmusára. Orv Hetil. 2021; 162(17): 652-667. Summary. Pediatric inflammatory multisystem syndrome (PIMS) is a rare complication of SARS-CoV-2 infection in children. PIMS is a severe condition, involving two or more organ systems. The symptoms overlap with Kawasaki disease, toxic shock syndrome and macrophage activation syndrome. PIMS patients should be treated in an intensive care unit or in an institution with an intensive care background, where cardiological care is also provided. The required specific immunotherapy depends on the clinical presentation. In this paper, after reviewing the relevant international literature, the authors make a recommendation for the diagnostic and therapeutic algorithm for PIMS. Orv Hetil. 2021; 162(17): 652-667

Development of a Novel LC-MS/MS Multi-Method for the Determination of Regulated and Emerging Food Contaminants Including Tenuazonic Acid, a Chromatographically Challenging <i>Alternaria</i> Toxin

The regulation of food contaminants in the European Union (EU) is comprehensive, and there are several compounds in the register or being added to the recommendation list. Recently, European standard methods for analysis have also been issued. The quick analysis of different groups of analytes in one sample requires a number of methods and the simultaneous use of various instruments. The aim of the present study was to develop a method that could analyze several groups of food contaminants: in this case, 266 pesticides, 12 mycotoxins, 14 alkaloid toxins, and 3 Alternaria toxins. The main advantage of the herein described approach over other methods is the simultaneous analysis of tenuazonic acid (TEA) and other relevant food contaminants. The developed method unites the newly published standard methods such as EN 15662:2018, EN 17194:2019, EN 17256:2019, EN 17425:2021, EN 17521:2021, which describes the analysis of both regulated and emerging contaminants. The developed method is based on a QuEChERS sample preparation, followed by LC-MS/MS analysis under alkaline mobile phase conditions. The pH of the aqueous eluent was set to 8.3, which resulted in baseline separation among ergot alkaloids and their corresponding epimers, a symmetric chromatographic peak shape for analyzing TEA and fit-for-purpose sensitivity for MS/MS detection in both positive and negative ionization modes. Those compounds, which possess the corresponding isotopically labeled internal standards (ISTD), allowed for direct quantification by the developed method and no further confirmation was necessary. This was proven by satisfactory analyses of a number of quality control (QC), proficiency test (PT), and validation samples

Safety and efficacy of a lipid emulsion containing a mixture of soybean oil, medium-chain triglycerides, olive oil, and fish oil

OBJECTIVES: Safety, tolerability, and efficacy of a novel lipid emulsion containing a mixture of soybean oil, medium-chain triglycerides, olive oil, and fish oil (SMOFlipid 20%) with reduced n-6 fatty acids (FA), increased monounsaturated and n-3 FA, and enriched in vitamin E were evaluated in premature infants compared with a soybean oil-based emulsion. PATIENTS AND METHODS: Sixty (30/30) premature neonates (age 3-7 days, gestational age </= 34 weeks, birth weights 1000-2500 g) received parenteral nutrition (PN) with either SMOFlipid 20% (study group) or a conventional lipid emulsion (Intralipid 20%, control group) for a minimum of 7 up to 14 days. Lipid supply started at 0.5 g . kg body weight(-1) . day(-1) on day 1 and increased stepwise (by 0.5 g) up to 2 g . kg body weight(-1) . day(-1) on days 4 to 14. Safety and efficacy parameters were assessed on days 0, 8, and 15 if PN was continued. RESULTS: Adverse events, serum triglycerides, vital signs, local tolerance, and clinical laboratory did not show noticeable group differences, confirming the safety of study treatment. At study end, gamma-glutamyl transferase was lower in the study versus the control group (107.8 +/- 81.7 vs 188.8 +/- 176.7 IU/L, P < 0.05). The relative increase in body weight (day 8 vs baseline) was 5.0% +/- 6.5% versus 5.1% +/- 6.6% (study vs control, not significant). In the study group, an increase in n-3 FA in red blood cell phospholipids and n-3:n-6 FA ratio was observed. Plasma alpha-tocopherol (study vs control) was increased versus baseline on day 8 (26.35 +/- 10.03 vs 3.67 +/- 8.06 mumol/L, P < 0.05) and at study termination (26.97 +/- 18.32 vs 8.73 +/- 11.41 mumol/L, P < 0.05). CONCLUSIONS: Parenteral infusion of SMOFlipid was safe and well tolerated and showed a potential beneficial influence on cholestasis, n-3 FA, and vitamin E status in premature infants requiring PN

Determination of Aminophosphonate Herbicides in Glutamate Loaded Spice Mix by LC-IDMS and Method Extension to Other Food Matrices

The accumulation of organophosphorus type herbicides has been observed worldwide in the environment (i.e. soil, water), together with their appearance in foods of plant origin. This paper reports a new liquid chromatography–isotope dilutiontandem mass spectrometric method (LC-IDMS) for the analysis of glufosinate (GLUF), glyphosate (GLY) and its main metabolite, aminomethylphosphonic acid (AMPA), in challenging food samples. Sample preparation is based on aqueous extraction with ethylenediaminetetraacetic acid solution, followed by solid-phase extraction (SPE) on mixed-mode cation exchange cartridges to remove matrix constituents before derivatization with -fluorenylmethoxycarbonyl chloride (FMOC-Cl). Derivatized samples were cleaned up on hydrophilic modified polymeric SPE cartridge. This two-step SPE supported sample preparation approach, and the LC-IDMS separation carried out in negative ionization mode resulted in fit-for-purpose recovery (81–118%) and precision (4–18%) in the validation of glutamate loaded spice mix, mushroom, maize and cherry samples. Amino acid content influencing FMOC derivatization efficiency was estimated with a HILIC-MS/MS setup. Mul- tiple reaction monitoring (MRM) was assisted with high-resolution (QTOF) accurate mass data on the FMOC-derivatized GLUF, GLY and AMPA standards. The limit of quantification (LOQ) was 0.005 mg/kg for all the three analytes. The method was successfully applied on quality control samples (oat and arugula) with fit-for-purpose accuracy (99–120%) and on other nineteen real samples, where GLY and AMPA were detected in the range between 0.005 and 0.069 mg/kg