Corrigendum to "Migration from RIA to LC-MS/MS for aldosterone determination: Implications for clinical practice and determination of plasma and urine reference range intervals in a cohort of healthy Belgian subjects" [Clin. Mass Spectrom. 9 (2018) 7-17].

[This corrects the article DOI: 10.1016/j.clinms.2018.06.002.].

Persistence of Isothiazolinones in Clothes After Machine Washing.

BACKGROUND: Sensitization to methylchloroisothiazolinone (MCI) and methylisothiazolinone (MI) is a worldwide problem. Washing machine detergents are suspected to cause cutaneous symptoms in highly sensitized patients. Little is known about the persistence of isothiazolinones in clothes that have been washed. OBJECTIVE: The aim of the study was to analyze the possible persistence of MI, MCI, benzisothiazolinone, and octylisothiazolinone in common fabrics after machine washing. METHODS: Different clothes (cotton, polyester, linen, and wool) were collected, and 4 types of wash were done (control, standard, standard + conditioner, and standard + double rinse). The samples were analyzed using ultrahigh-performance liquid chromatography. RESULTS: The results showed that the concentrations of isothiazolinones were very low, independent of the type of material or wash. The highest levels were found in the control wash (hand wash), reaching a maximum of 0.4 ppm in the linen. CONCLUSIONS: Our findings suggest that it is not necessary to recommend that patients sensitized to MI avoid isothiazolinones in machine detergents or fabric conditioners or to double rinse. However, after using the detergent for hand washing (the control in our study), there may remain sufficient concentrations of isothiazolinones in clothes to produce symptoms in highly sensitized patients.

Determination of Cystatin C in human urine by isotope dilution tandem mass spectrometry.

This work presents the development of a methodology for the accurate and precise quantification of the renal biomarker Cystatin C in human urine by Isotope Dilution Mass Spectrometry (IDMS). The procedure is based on the addition of a known quantity of the proteotypic peptide ALDFAVG*EYNK labelled with 13C2-glycine to the urine sample followed by protein hydrolysis using trypsin. Then, preconcentration and purification of the isotope diluted peptide was carried out by a selective monoclonal antibody bound to magnetic beads and final measurement was done after injection of the sample in a HPLC-MS/MS triple quadrupole instrument. The isotopic distribution of the isotope diluted proteotypic peptide was measured by low resolution selected reaction monitoring. Using this aquisition mode, the bandpass of the first quadrupole was widened (FWHM =13 u) so the whole isotopic clusters for both the natural abundance and the labelled peptides entered the collision cell. The proposed acquisition mode provided similar accuracy and precision than the regular SRM mode (FWHM =0.7 u) but a higher sensitivity was observed. The purification of the sample by antibody based enrichment of the target peptide was shown to remove interfering compounds more efficiently in comparison with a sample purification based on semipreparative liquid chromatography. Using 5 ng of the labelled peptide it was possible to quantify Cystatin C in human urine in patients with normal and impaired renal function. Recoveries from 100 to 104% were obtained in samples containing from 90 to 700 µg L-1 of Cystatin C with relative standard deviations from 0.5 to 6%. The stability of Cystatin C in urine samples was evaluated under different storage conditions showing that only when the urine samples were stored at room temperature during more than 10 days, a significant degradation of Cystatin C was observed.

Assessment of 22 inorganic elements in human amniotic fluid: a cross-sectional study conducted in Canary Islands (Spain).

We conducted a cross-sectional study in the Hospital Universitario de Canarias (Tenerife, Canary Islands, Spain) measuring 22 inorganic elements in amniotic fluid (AF) samples obtained from 65 pregnant women. ICP-MS was used for quantification of inorganic elements. Newborn parameters at delivery were all within the normal range. Concentrations of all elements were detected in measurable amounts in AF. The concentration of elements was similar to those reported in the literature. The concentrations of the most dangerous heavy metals - Cd, Cr, Ni, Hg and Pb - were lower than those reported by other authors. This study demonstrates that toxic inorganic elements pass into and accumulate in AF. The presence of such pollutants in contact with developing embryos from the intrauterine period could exert adverse health effects that deserve future investigations.

Body burden of toxic metals and rare earth elements in non-smokers, cigarette smokers and electronic cigarette users.

Smoking is considered an important source for inorganic elements, most of them toxic for human health. During the last years, there has been a significant increase in the use of e-cigarettes, although the role of them as source of inorganic elements has not been well established. A cross-sectional study including a total of 150 subjects from Brasov (Romania), divided into three groups (non-smokers, cigarette smokers and electronic cigarettes smokers) were recruited to disclose the role of smoking on the human exposure to inorganic elements. Concentration of 42 elements, including trace elements, elements in the ATSDR's priority pollutant list and rare earth elements (REE) were measured by ICP-MS in the blood serum of participants. Cigarette smokers showed the highest levels of copper, molybdenum, zinc, antimony, and strontium. Electronic cigarette (e-cigarette) users presented the highest concentrations of selenium, silver, and vanadium. Beryllium, europium and lanthanides were detected more frequently among e-cigarette users (20.6%, 23.5%, and 14.7%) than in cigarette smokers (1.7%, 19.0%, and 12.1%, respectively); and the number of detected REE was also higher among e-cigarette users (11.8% of them showed more than 10 different elements). Serum levels of cerium and erbium increased as the duration of the use of e-cigarettes was longer. We have found that smoking is mainly a source of heavy metals while the use of e-cigarettes is a potential source of REE. However, these elements were detected at low concentrations.

Pattern of blood concentrations of 47 elements in two populations from the same geographical area but with different geological origin and lifestyles: Canary Islands (Spain) vs. Morocco.

The Canary Islands are one of the outermost regions of the European Union (EU), which are located barely 100 km from the coasts of Morocco. Although these islands are located in Africa, the degree of socioeconomic development and lifestyle in this archipelago is comparable to that of any other region of Europe. It is well established that the main determinants of human exposure to elements have to do both, with their place of residence and with habits related to their lifestyle. For this reason, we wanted to study the pattern of contamination by elements of these two populations so geographically close, but so different both in their lifestyle, and the geological origin of the territory where they live. Thus, we have determined the blood concentrations of 47 elements (including 25 rare earth elements (REE) and other minority elements (ME) widely employed in the hi-tech industry) in a paired sample of Moroccans (n = 124) and Canary Islands inhabitants (n = 120). We found that the levels of iron, selenium, zinc, arsenic, cadmium, strontium, and specially lead, were significantly higher in Moroccans than in Canarians, probably due to the intensive mining activity in this country. We also found significantly higher levels of the sum of REE and ME in Moroccans than in Canarians, possibly related to the inappropriate management of e-waste in this country. On the other hand, in the inhabitants of the Canary Islands we found higher levels of manganese, probably related to a higher degree of exposure to heavy traffic and exposure to Saharan dust of the people living in this region, and niobium and bismuth, probably related to the higher economic development in these islands. Our results indicate that the vicinity of both territories is not a major determinant of each other's contamination.

Occurrence of 44 elements in human cord blood and their association with growth indicators in newborns.

There is growing concern about environmental pollution produced by elements, including "emerging" contaminants, such as rare earth elements (REE) and other trace elements (TE), which are extensively and increasingly employed in the manufacture of consumer electronics. Previous research has shown that prenatal exposure to some elements (mainly heavy metals) may be associated with decreased fetal growth and other adverse birth outcomes. Recent studies have also shown that environmental exposure to REE and TE may be related to adverse effects on human health. This cross-sectional study, which included nearly 92% of the births in 2016 in La Palma (Canary Islands, Spain; n = 471), aimed to evaluate the potential adverse health effects exerted by a wide range of elements on newborns. We quantified the levels of 44 elements (including 26 REE and TE) in their umbilical cord blood. Our results showed low or very low levels of most elements. We found an inverse association between antimony (Sb) and birth weight (Spearman's r = -0.106, p = 0.021). A similar trend was observed between nickel (Ni) and birth weight and between chromium (Cr) and birth length, although in this case the significance was borderline. Bismuth appeared as a risk factor for having a birth weight below the tenth percentile in the univariate (OR = 3.30; 95% CI = 1.25-8.78; p = 0.017) and multivariate analyses (OR = 5.20; 95% CI = 1.29-20.91; p = 0.020). When assessing the effect of element mixtures, the sum of Cr, Ni, and Sb appeared as a risk factor for having a birth weight below the tenth percentile in the univariate (OR = 2.41; 95% CI = 1.08-5.35; p = 0.031) and multivariate analyses (OR = 3.84; 95% CI = 1.42-10.39; p = 0.008). Our findings suggest that some inorganic elements-isolated or in mixture-are associated to a lower fetal growth. Additional research is needed to understand the role of inorganic pollutants on fetal development.

Blood levels of toxic metals and rare earth elements commonly found in e-waste may exert subtle effects on hemoglobin concentration in sub-Saharan immigrants.

Pollution by heavy metals and more recently by rare earth elements (REE) and other minor elements (ME) has increased due in part to their high use in technological and electronic devices. This contamination can become very relevant in those sites where e-waste is improperly processed, as it is the case in many countries of the African continent. Exposure to some toxic elements has been associated to certain hematological disorders, specifically anemia. In this study, the concentrations of 48 elements (including REE and other ME) were determined by ICP-MS in whole blood samples of sub-Saharan immigrants with anemia (n=63) and without anemia (n=78). We found that the levels of Fe, Cr, Cu, Mn, Mo, and Se were significantly higher in the control group than in the anemia group, suggesting that anemia was mainly due to nutritional deficiencies. However, since other authors have suggested that in addition to nutritional deficiency, exposure to some elements may influence hemoglobin levels, we wanted to explore the role of a broad panel of toxic and "emerging" elements in hemoglobin deficiency. We found that the levels of Ag, As, Ba, Bi, Ce, Eu, Er, Ga, La, Nb, Nd, Pb, Pr, Sm, Sn, Ta, Th, Tl, U and V were higher in anemic participants than in controls. For most of these elements an inverse correlation with hemoglobin concentration was found. Some of them also correlated inversely with blood iron levels, pointing to the possibility that a higher rate of intestinal uptake of these could exist in relation to a nutritional deficiency of iron. However, the higher levels of Pb, and the group of REE and other ME in anemic participants were independent of iron levels, pointing to the possibility that these elements could play a role in the development of anemia.

Persistent organic pollutants and risk of diabetes and obesity on healthy adults: Results from a cross-sectional study in Spain.

Environmental exposure to persistent organic pollutants (POPs) has been reported to be relevant in the population of the Canary Islands (Spain), especially that of organochlorine pesticides. On the other hand, the population of this archipelago presents a high prevalence of type 2 diabetes (T2D), and it has been recently reported that environmental chemical contamination could play a role in the development of this disease. We performed a cross-sectional study in a representative sample from this archipelago to evaluate whether serum levels of selected POPs could be considered as risk factors for diabetes in this population. Serum levels of 30 POPs were determined in 429 adults (9.3% with T2D). We found that serum levels of p,p'-DDE (DDE), PCB-153 and PCB-118 were significantly higher among subjects having diabetes than in non-diabetic subjects (p=0.001, p=0.046, and p<0.0001, respectively). We observed a positive correlation between serum p,p'-DDE and glucose levels. Serum p,p'-DDE was identified as a risk factor for diabetes in univariate analysis in the whole series, and it remained as an independent risk factor for diabetes in subjects with serum glucose <126mg/dL (multivariate analysis, Exp(B)=1.283, CI 95% (1.023-1.611), p=0.031). Those normoglycemic subjects that are most exposed to p,p'-DDE (95th percentile: serum p,p'-DDE>5µg/L) seem to be those people at higher risk. Our results showed that p,p'-DDE levels were significantly higher among subjects having diabetes. These findings should be considered by public health Authorities to implement measures devoted to minimize human exposure to pollutants that could be harmful to the population.

Study of the influencing factors of the blood levels of toxic elements in Africans from 16 countries.

Africa's economy is growing faster than any other continent and it has been estimated that the middle class in Africa now exceeds 350 million people. This has meant a parallel increase in the importation of consumer goods and in the implementation of communication and information technologies (ICT), but also in the generation of large quantities of e-waste. However, inadequate infrastructure development remains a major constraint to the continent's economic growth and these highly toxic residues are not always adequately managed. Few studies have been conducted to date assessing the possible association between socioeconomic development factors, including e-waste generation, and blood levels of inorganic elements in African population. To disclose the role of geographical, anthropogenic, and socioeconomic development determinants on the blood levels of Ag, Al, As, Be, Cd, Co, Cr, Hg, Ni, Pb, Sb, and V -all of them frequently found in e-waste-, an immigrant population-based study was made including a total of 245 subjects from 16 countries recently arrived to the Canary Islands (Spain). Women presented higher levels of blood elements than men, and Northern Africans (Moroccans) were the most contaminated. People from low-income countries exhibited significantly lower blood levels of inorganic elements than those from middle-income countries. We found a significant association between the use of motor vehicles and the implementation of information and communication technologies (ICT) and the level of contamination. Immigrants from the countries with a high volume of imports of second-hand electronic equipment, telephone and internet use had higher levels of inorganic elements. In general terms, the higher level of economic development the higher the blood levels of inorganic pollutants, suggesting that the economic development of Africa, in parallel to e-waste generation and the existence of informal recycling sites, have directly affected the level of contamination of the population of the continent.

Multicenter Evaluation of Cystatin C Measurement after Assay Standardization.

BACKGROUND: Since 2010, a certified reference material ERM-DA471/IFCC has been available for cystatin C (CysC). This study aimed to assess the sources of uncertainty in results for clinical samples measured using standardized assays. METHODS: This evaluation was performed in 2015 and involved 7 clinical laboratories located in France and Belgium. CysC was measured in a panel of 4 serum pools using 8 automated assays and a candidate isotope dilution mass spectrometry reference measurement procedure. Sources of uncertainty (imprecision and bias) were evaluated to calculate the relative expanded combined uncertainty for each CysC assay. Uncertainty was judged against the performance specifications derived from the biological variation model. RESULTS: Only Siemens reagents on the Siemens systems and, to a lesser extent, DiaSys reagents on the Cobas system, provided results that met the minimum performance criterion calculated according to the intraindividual and interindividual biological variations. Although the imprecision was acceptable for almost all assays, an increase in the bias with concentration was observed for Gentian reagents, and unacceptably high biases were observed for Abbott and Roche reagents on their own systems. CONCLUSIONS: This comprehensive picture of the market situation since the release of ERM-DA471/IFCC shows that bias remains the major component of the combined uncertainty because of possible problems associated with the implementation of traceability. Although some manufacturers have clearly improved their calibration protocols relative to ERM-DA471, most of them failed to meet the criteria for acceptable CysC measurements.

Simultaneous quantification of 49 elements associated to e-waste in human blood by ICP-MS for routine analysis.

Health risks concerns related to e-waste contamination are increasing all over the world, and especially in developing countries. We have developed an easy, quick, and robust method for the quantification of 49 elements associated to electronic consumer products and their e-wastes in human blood. An aliquot of blood (130 µL) is simply diluted using an alkaline solution, and the elements are simultaneously quantified at the picogram-per-milliliter level without the need of clean-up steps. The accuracy, precision, linearity and limit of quantification (LOQ) of the method were assessed. Recovery values at concentration levels between 0.010 and 5 ng mL-1 were studied. A range of 89-118% and a range of 87-128% for REE and toxic heavy elements was found respectively. The relative standard deviations (RSD) calculated were lower than 8% for the most elements. The limits of quantification (LOQ) were found to be lower than 0.04 ng mL-1 for toxic heavy elements and 0.06 ng mL-1 for the REE with some few exceptions in both cases. The validity of the proposed methodology was assessed by analyzing a certified human blood reference material with known concentrations of several elements. The proposed method is suitable for routine use in biomonitoring studies.

Determination of Cystatin C in human serum by isotope dilution mass spectrometry using mass overlapping peptides.

We propose a peptide-based isotope dilution mass spectrometry approach for Cystatin C determination in human serum samples, a clinical marker for renal status for which backup by a mass spectrometry based primary method has been missing so far. In contrast to common protocols, the isotope labelled version of the proteotypic signature peptide is designed such as keeping the isotopic difference as little as possible with respect to the peptide released from the protein. Peptides labelled in two (13)C atoms are added to the serum samples just before proteolysis. After two steps of chromatographic purification the sample is measured by selected reaction monitoring using a LC-MS/MS. Resolution of the first quadrupole is reduced to transmit the whole parent ion cluster to the collision cell for monitoring accurate isotopic distributions of the molecular fragments. Molar fractions of labelled and natural abundance peptides are directly obtained from the experimental mass spectra of the in-cell fragment ions. Thus, the natural abundance protein concentration is obtained from the fragment-ion spectrum of the sample without resorting to extra calibration runs. Applicability of the approach is demonstrated by the measurement of the serum concentration of Cystatin C in Reference Material ERM R-DA471/IFCC and real samples. BIOLOGICAL SIGNIFICANCE: Cystatin C is used as an alternative marker instead of, or in combination with creatinine for non-invasive determination of glomerular filtration rates. Advantages advocating in favour of Cystatin C in diagnosis of chronic kidney diseases are the lower variability of its serum level and, particularly, virtual independence on sex, age and muscle mass. However, in order to capitalize, accuracy of measurement has to be in proportion with the predictive power of the marker. Though there are label-free methods available for screening purposes or high-throughput analysis, achieving high levels of reliability and accuracy in quantitative proteomics takes reference to isotope labelled materials. Present routine assays (mainly nephelometry, turbidimetry and ligand-binding assays) are known to leave improvement to be desired in that respect. Absolute quantification based on enzymatic signature-peptides provides a method principle establishing traceability to the International System of Units on the level of primary methods. The kind of technique is capable, by this way, of high accuracy value-assignment to matrix materials needed for calibration of present routine assays, where not completely replacing them. Cystatin C measurement by isotope dilution mass spectrometry is developed in this study with the aim of making available this tool to support diagnostics of kidney function in the same way.

Simultaneous determination of seven ß2-agonists in human and bovine urine by isotope dilution liquid chromatography-tandem mass spectrometry using compound-specific minimally 13C-labelled analogues.

Seven ß2-agonist (clenproperol, clenbuterol, salbutamol, bronbuterol, ractopamine, clenpenterol and clencyclohexerol) were determined simultaneously in human and bovine urine by isotope dilution LC-ESI-MS/MS in a triple quadrupole instrument. The method is based on the application of multiple linear regression in combination with compound-specific minimally 13C-labelled analogues. Additionally, the increase of the bandpass of the first quadrupole during the selected reaction monitoring (SRM) measurement procedure allowed the simultaneous quantification of the seven compounds at sub ngg-1 levels in a single chromatogram without resorting to a methodological calibration graph. Recovery values at concentration levels between 5.0 and 0.05ngg-1 ranged from 95 to 110% in fortified bovine urine and from 91 to 108% in human urine, with relative standard deviations lower than 5% except for salbutamol and ractopamine. The proposed methodology was validated by analyzing the certified reference material BCR-503 (lyophilized bovine urine) certified for clenbuterol and salbutamol. The limits of detection (LOD) for a sample volume of 10mL of both human and bovine urine was found to be lower than 0.012ngg-1 for all compounds, except to salbutamol in bovine urine which was of 0.029ngg-1. The use of compound-specific isotopically labelled analogues minimally labelled in 13C minimized the occurrence of isotope effects and corrected for matrix effects during ESI ionization and can be efficiently applied for the quantification of ultra-trace concentrations of ß2-agonists in human and bovine urine.

Determination of the enrichment of isotopically labelled molecules by mass spectrometry.

A general method for the determination of the enrichment of isotopically labelled molecules by mass spectrometry (MS) is described. In contrast to other published procedures, the method described here takes into account and corrects for measurement errors such as the contribution at M - 1 due to loss of hydrogen or lack of spectral resolution and provides an uncertainty value for the determined enrichment. The general procedure requires the following steps: (1) evaluation of linearity in the mass spectrometer by injecting the natural abundance compound at different concentration levels, (2) determination of the purity of the mass cluster using the natural abundance analogue, (3) calculation of the theoretical isotope composition of the labelled compound using different tentative isotope enrichments, (4) calculation of 'convoluted' isotope distributions for the labelled compound taking into account the purity of the mass cluster determined with the natural abundance analogue and (5) comparison of the isotope distributions measured for the labelled compound with those calculated for different isotope enrichments using linear regression. The method was applied to a series of commercially available (13)C- and (2)H-labelled compounds and to a suite of singly (13)C-labelled ß2-agonist prepared in-house both by gas chromatography (GC)-MS, GC-tandem MS (MS/MS) and liquid chromatography-MS/MS with satisfactory results. It was observed that the main uncertainty source for the isotope enrichment was the uncertainty in the purity of the measured cluster as determined with the natural abundance compound.

Development of an isotope dilution GC-MS procedure for the routine determination of creatinine in complex serum samples.

The accurate determination of creatinine in serum is essential for the diagnosis and treatment of kidney diseases. The determination of serum creatinine in clinical laboratories is routinely carried out by the Jaffe method or by enzymatic methods that may suffer from interferences. So, the development of reliable, fast and interference-free routine methods for complex serum samples is required. A fast method using isotope dilution mass spectrometry (IDMS) and gas chromatography mass spectrometry (GC-MS) was developed using minimally (13)C labeled creatinine analogs, multiple linear regression and rapid derivatization of creatinine with acetylacetone in 2 min by using focused microwave technology. The proposed method was validated with the analyses of two Certified Reference Materials (ERM-DA252a and ERM-DA253a) and compared with the Jaffe and enzymatic methods when analyzing real serum samples containing variable levels of bilirubin The proposed method is capable of providing accurate serum creatinine concentrations in less than 45 min from sample arrival to full data treatment and can be an alternative routine procedure for creatinine determinations in complex serum samples.

Overcoming matrix effects in electrospray: quantitation of ß-agonists in complex matrices by isotope dilution liquid chromatography-mass spectrometry using singly (13)C-labeled analogues.

In this work, the implementation of isotope dilution mass spectrometry (IDMS) using minimal labeling and isotope pattern deconvolution (IPD) is evaluated as a strategy for the minimization of matrix effects during trace determination of ß2-agonists in complex matrices by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). First, the parameters affecting the measurement of isotopic composition of organic compounds by liquid chromatography electrospray ionization high resolution mass spectrometry with a time-of-flight analyzer were evaluated using as a case of study three different ß2-agonists: clenbuterol, clenproperol and brombuterol. Then, a calibration graph-free IDMS methodology was evaluated in order to overcome matrix effects in LC-ESI-MS in complex samples. In this procedure singly (13)C-labeled analogues of clenbuterol, clenproperol and brombuterol were employed in combination with IPD. Using this approach accurate and precise results were obtained in the simultaneous quantification of ß2-agonists in human urine and bovine liver, even at the sub ngg(-1) and particularly in spite of the previously reported matrix effects. Recovery rates in the range of 97-114% in fortified human urine and from 95% to 111% in fortified bovine liver were obtained with RSD (%) of independent recovery experiments always lower than 6%. These results demonstrate that the proposed methodology based on the use of (13)C1-labeled standards and IPD is a reliable approach for accurate LC-MS quantitation of small molecules and compatible with full-scan high-resolution mass spectrometry.

Development of a routine method for the simultaneous confirmation and determination of clenbuterol in urine by minimal labeling isotope pattern deconvolution and GC-EI-MS.

A novel and fast routine method for the simultaneous determination and confirmation of clenbuterol in bovine and human urine samples by gas chromatography electron ionization mass spectrometry (GC-EI-MS) has been developed. The method employs isotope dilution mass spectrometry (IDMS) and is based on a combination of minimal labeling (a single (13)C label in the molecule) and isotope pattern deconvolution (IPD). This new methodology does not require the construction of a methodological calibration graph, and was compared with the classical IDMS procedure employed in clenbuterol analysis based on the use of a deuterated compound as internal standard (d(9)-clenbuterol) and a calibration curve. The sample preparation consists of simple extraction with dichloromethane, which was dried and derivatized with chloro(chloromethyl)dimethylsilane, generating a cyclic dimethylsilamorpholine (DMS) derivative suitable for GC(EI)MS detection and identification. This compound produces five intense ions in the electron ionization source, which allow the presence of clenbuterol to be confirmed in just one analysis, as demanded by European Union directives. The accuracy of the method was studied by performing recovery experiments at different concentration levels (from 0.3 to 5 ng g(-1)) in 5 mL bovine urine samples using two labeled compounds: an in-house-synthesized (13)C(1)-clenbuterol and a commercially available d(9)-clenbuterol. The detection limit of the method in human urine was 0.050 ng g(-1) with a sample volume of 10 mL, and is thus suitable for antidoping control purposes. Finally, the (13)C(1)-clenbuterol standard was employed for the determination of clenbuterol in two reference materials, BCR-503 and BCR-504 (lyophilized bovine urine). The concentrations obtained were in agreement with the certified values, with a reproducibility of below 1% RSD.

Evaluation of minimal 13C-labelling for stable isotope dilution in organic analysis.

A procedure for Stable Isotope Dilution Analysis in molecular Mass Spectrometry which does not require a methodological calibration graph and can be applied in combination with minimal labelling has been evaluated. This alternative approach is based on the determination of the molar fractions for each pure isotope pattern (natural abundance or labelled) contributing to the isotope pattern observed in the mixture of natural abundance and labelled molecules by multiple linear regression. Two labelled compounds, (13)C(1)-labelled or (13)C(6)-labelled phenol, were compared to study the influence of the number of (13)C atoms in the labelled molecule. The procedure was evaluated by comparing the results obtained for the determination of phenol in NIST 1584 CRM by GC-EI-MS using the classical isotope dilution calibration procedure and the new procedure based on multiple linear regression of isotope patterns without a calibration graph. The results obtained using the proposed procedure agreed well with both the certified values and those obtained using the classical Isotope Dilution Mass Spectrometry (IDMS) calibration procedures. For the evaluated procedure, a full uncertainty budget determination has been developed taking into account all uncertainty sources, including those derived from the uncertainties in the isotope patterns of the natural and labelled compounds. The measurements with the (13)C(1)-labelled phenol provided lower propagated uncertainties in comparison to the use of (13)C(6)-labelled phenol.