Enantiomeric methadone quantitation on real post-mortem dried matrix spots samples: Comparison of liquid chromatography and supercritical fluid chromatography coupled to mass spectrometry.

This study describes two bioanalytical methods for the quantitation of the two methadone enantiomers in dried matrix spots using high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) and high performance supercritical chromatography tandem mass spectrometry (HPSFC-MS/MS). Dried matrix spots were obtained by spotting 10 µL of each sample fluid on a Whatman paper. Methadone and its main metabolite, EDDP, were extracted with 100 µL methanol and subsequently injected into the LC-MS/MS and SFC-MS/MS systems. Enantiomeric separation was achieved with AGP-column for the LC conditions and with Chiralpak IH-3 in SFC. The two methods were fully validated and 93 post-mortem samples were analysed with both analytical methods. Results from validation parameters and results obtained for all post-mortem samples were compared with a significant spearman correlation of r <sub>s</sub> = 0.9978 for R-methadone and r <sub>s</sub> = 0.9981 for S-methadone. The LC method provided better results in terms of uncertainty, retention factor and resolution, whereas SFC provides better sensitivity, with lower LOD. Median R-/S-methadone ratio in peripheral blood was found equal to 1.60 (N = 32), varying from 0.79 to 4.23. The reported values were in good agreement with previously published results. Based on the results obtained here, SFC-MS/MS can be considered a reliable alternative to the widely used LC-MS/MS for the quantitation of methadone enantiomers in bioanalysis and should be evaluated for other bioanalytical methods. Both methods can be easily and quickly used in toxicological routine analysis for the methadone quantitation in human fluids matrices, even if considering that the polysaccharide coated column IH-3 used in SFC does not allow the enantiomeric EDDP separation

Isotonitazene: Fatal intoxication in three cases involving this unreported novel psychoactive substance in Switzerland.

The paper describes the first three deaths reported in Europe involved in isotonitazene consumption, a potent benzimidazole derivate opioid consumed in the recreational drug scene. Isotonitazene powder and purity determination was performed on the sample collected in the first death scene by NMR, HRMS, GC-FTIR, ATR-FTIR and GC-MS. Isotonitazene purity was determined by GC-MS analysis and proton NMR, and was defined to be above 95 % and 98 %, respectively. Quantification of isotonitazene in biological samples was performed using a targeted analysis based on SPE extraction and ultra-high performance liquid chromatography tandem mass spectrometry. The isotonitazene median concentration in femoral whole blood was 1.20ng/mL. Isotonitazene concentration in hair was similar or even lower compared to that seen in fentanyl abusers. Isotonitazene distribution in tissues converges in the brain, lungs and heart, respectively. Surprisingly, isotonitazene concentration in liver is the lowest measured for all tissues and fluids analyzed. Based on circumstantial evidence, autopsy findings and the results of the toxicological analysis, the medical examiner concluded that the cause of all three deaths was an acute intoxication with isotonitazene. Since isotonitazene toxic concentration levels are very low, the consumption of this new psychoactive drug is a real hazard for human health

The Iowa Homemaker vol.3, no.8

Table of Contents The Goal of Home Management Courses at Iowa State by Ruth M. Lindquist, page 3 Home Economics Venture Upon “India’s Coral Strand” by Marcia E. Turner, page 4 The Bell Recipe File by Opal Wind, page 4 Carving the Turkey by Viola M. Bell, page 5 An Interview With Scottish Hockey Coach by Lucile Barta, page 5 Two and One-half Miles Saved a Day by Grata Thorne, page 6 Cranberries for Thanksgiving by Katherine Goeppinger, page 7 Who is Responsible for the Child - by “A Bachelor”, page 8 The Evolution of Home Economics at Iowa State by Ruth Elaine Wilson, page 9 Why is a Mulligan? by “Joe Baggs”, page 10 Turning the Corners Up by Laura E. Bublitz, page 10 The Sport of Amateur Housekeeping by Anna Jacobson, page 11 Who’s There and Where by Helen I. Putnam, page 12 Dressing the Homemaker by Margaret Dix, page 1

The Iowa Homemaker vol.3, no.2

Table of Contents It Is Not Always May by Maybelle A. Payton, page 1 The Why of a Home Economics Course by Florence Busse, page 2 Why I Came to Iowa State compiled by Clara Jordan, page 2 Picnic Ingredients by Grata Thorn, page 3 Iowa State Women Attend Voters’ Convention by Eleanor Murray and Jeanette Beyer, page 4 A Modern Version of the Hope Box by N. Beth Bailey, page 5 A Summer Living Room by Mildred Boyt, page 7 Canning Early Fruits and Vegetables by Helen G. Lamb, page 8 Hazards of Bird Life by J. E. Guthrie, page 9 Nile Styles by Harriett Schleiter, page 10 Shall Mother Have a Vacation? by Eda Lord Murphy, page 10 The Fallacy of An Expensive Standard of Living by Claude L. Benner, page 11 What Shall We Take? by Lucille Barta, page 12 Who’s There and Where by Helen Reidy, page 1

Recent advances of metabolomics in plant biotechnology

Biotechnology, including genetic modification, is a very important approach to regulate the production of particular metabolites in plants to improve their adaptation to environmental stress, to improve food quality, and to increase crop yield. Unfortunately, these approaches do not necessarily lead to the expected results due to the highly complex mechanisms underlying metabolic regulation in plants. In this context, metabolomics plays a key role in plant molecular biotechnology, where plant cells are modified by the expression of engineered genes, because we can obtain information on the metabolic status of cells via a snapshot of their metabolome. Although metabolome analysis could be used to evaluate the effect of foreign genes and understand the metabolic state of cells, there is no single analytical method for metabolomics because of the wide range of chemicals synthesized in plants. Here, we describe the basic analytical advancements in plant metabolomics and bioinformatics and the application of metabolomics to the biological study of plants

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

A one-year monitoring of nicotine use in sport: frontier between potential performance enhancement and addiction issues.

Tobacco consumption is a global epidemic responsible for a vast burden of disease. With pharmacological properties sought-after by consumers and responsible for addiction issues, nicotine is the main reason of this phenomenon. Accordingly, smokeless tobacco products are of growing popularity in sport owing to potential performance enhancing properties and absence of adverse effects on the respiratory system. Nevertheless, nicotine does not appear on the 2011 World Anti-Doping Agency (WADA) Prohibited List or Monitoring Program by lack of a comprehensive large-scale prevalence survey. Thus, this work describes a one-year monitoring study on urine specimens from professional athletes of different disciplines covering 2010 and 2011. A method for the detection and quantification of nicotine, its major metabolites (cotinine, trans-3-hydroxycotinine, nicotine-N'-oxide and cotinine-N-oxide) and minor tobacco alkaloids (anabasine, anatabine and nornicotine) was developed, relying on ultra-high pressure liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-TQ-MS/MS). A simple and fast dilute-and-shoot sample treatment was performed, followed by hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS) operated in positive electrospray ionization (ESI) mode with multiple reaction monitoring (MRM) data acquisition. After method validation, assessing the prevalence of nicotine consumption in sport involved analysis of 2185 urine samples, accounting for 43 different sports. Concentrations distribution of major nicotine metabolites, minor nicotine metabolites and tobacco alkaloids ranged from 10 (LLOQ) to 32,223, 6670 and 538 ng/mL, respectively. Compounds of interest were detected in trace levels in 23.0% of urine specimens, with concentration levels corresponding to an exposure within the last three days for 18.3% of samples. Likewise, hypothesizing conservative concentration limits for active nicotine consumption prior and/or during sport practice (50 ng/mL for nicotine, cotinine and trans-3-hydroxycotinine and 25 ng/mL for nicotine-N'-oxide, cotinine-N-oxide, anabasine, anatabine and nornicotine) revealed a prevalence of 15.3% amongst athletes. While this number may appear lower than the worldwide smoking prevalence of around 25%, focusing the study on selected sports highlighted more alarming findings. Indeed, active nicotine consumption in ice hockey, skiing, biathlon, bobsleigh, skating, football, basketball, volleyball, rugby, American football, wrestling and gymnastics was found to range between 19.0 and 55.6%. Therefore, considering the adverse effects of smoking on the respiratory tract and numerous health threats detrimental to sport practice at top level, likelihood of smokeless tobacco consumption for performance enhancement is greatly supported

Biological control of Rhizoctonia solani AG 2-2IIIB by Bacillus subtilis metabolites

Celem doświadczenia było określenie aktywności metabolitów produkowanych przez bakteryjny szczep Bacillus subtilis wobec izolatu Rhizoctonia solani należącego do grupy AG 2-2IIIB. Antagonistyczne właściwości metabolitów B. subtilis były oceniane w kulturach płytkowych na podłożu Czapka po 6, 24 i 48 godzinach hodowli w temperaturze 30 i 37ºC. Wpływ metabolitów wytwarzanych przez B. subtilis na wzrost R. solani AG 2-2IIIB przedstawiono w postaci współczynnika tempa wzrostu liniowego grzyba. Uzyskane wyniki wykazały, że na fungistatyczną aktywność metabolitów B. subtilis wobec R. solani AG 2-2IIIB wpływa zarówno czas, jak i temperatura inkubacji bakterii. Wzrost grzybni był najmocniej hamowany przez metabolity uzyskane po 6-godzinnej hodowli w temp. 37ºC.The aim of the experiment was to determine the activity of metabolites produced by bacteria Bacillus subtilis Kg against fungus isolate of Rhizoctonia solani ID 105 belonging to the anastomosis group AG 2-2IIIB. The antagonist properties of B. subtilis Kg metabolites were evaluated with a culture-plate method on Czapek growth media for B. subtilis Kg cultures after 6, 24 and 48 hours of culture at temp. 30 and 37°C. The impact of B. subtilis Kg metabolites on the growth of R. solani AG 2-2IIIB was shown as a growth rate index of the fungus. The results showed that on the fungistatic activity of the metabolites of B. subtilis against R. solani AG 2-2IIIB affects both the time and temperature of the bacterial culture. Mycelial growth was most strongly inhibited by the metabolites obtained after 6 hours cultivation at 37°C

SARM-S4 and metabolites detection in sports drug testing: a case report.

Recently, pharmaceutical industry developed a new class of therapeutics called Selective Androgen Receptor Modulator (SARM) to substitute the synthetic anabolic drugs used in medical treatments. Since the beginning of the anti-doping testing in sports in the 1970s, steroids have been the most frequently detected drugs mainly used for their anabolic properties. The major advantage of SARMs is the reduced androgenic activities which are the main source of side effects following anabolic agents' administration. In 2010, the Swiss laboratory for doping analyses reported the first case of SARMs abuse during in-competition testing. The analytical steps leading to this finding are described in this paper. Screening and confirmation results were obtained based on liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses. Additional information regarding the SARM S-4 metabolism was investigated by ultra high-pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometer (UHPLC-QTOF-MS)