Tiphaine Choteau

Mass Balance Method for the SI Value Assignment of the Purity of Organic Compounds

A mass balance method is described for determining the mass fraction of the main component of a high purity organic material. The resulting assigned value is established to be traceable to the SI and can be determined with a small associated measurement uncertainty. Pure organic materials with values and uncertainties determined in this way are necessary as primary calibrators of reference measurement systems in order to underpin the metrological traceability of routine measurement results. The method has been applied to materials in which the main components were respectively theophylline, digoxin, 17β-estradiol, and aldrin. Its performance has been validated in international comparisons coordinated by the BIPM and is in principle applicable to a wide structural range of stable, nonvolatile organic compounds. It has been successfully applied to mass fraction assignments when the main component is present in the range of (950-1000) mg/g and can achieve associated standard uncertainties ranging from 0.5 mg/g (for high purity materials or those containing well-characterized, stable minor components) to 2 mg/g (materials with a significant number or variety of impurities). It is in principle equally applicable to materials with a smaller mass fraction content of the main component.

Methane Standards Made in Whole and Synthetic Air Compared by Cavity Ring Down Spectroscopy and Gas Chromatography with Flame Ionization Detection for Atmospheric Monitoring Applications

There is evidence that the use of whole air versus synthetic air can bias measurement results when analyzing atmospheric samples for methane (CH4) and carbon dioxide (CO2). Gas chromatography with flame ionization detection (GC-FID) and wavelength scanned-cavity ring down spectroscopy (WS-CRDS) were used to compare CH4 standards produced with whole air or synthetic air as the matrix over the mole fraction range of 1600-2100 nmol mol(-1). GC-FID measurements were performed by including ratios to a stable control cylinder, obtaining a typical relative standard measurement uncertainty of 0.025%. CRDS measurements were performed using the same protocol and also with no interruption for a limited time period without use of a control cylinder, obtaining relative standard uncertainties of 0.031% and 0.015%, respectively. This measurement procedure was subsequently used for an international comparison, in which three pairs of whole air standards were compared with five pairs of synthetic air standards (two each from eight different laboratories). The variation from the reference value for the whole air standards was determined to be 2.07 nmol mol(-1) (average standard deviation) and that of synthetic air standards was 1.37 nmol mol(-1) (average standard deviation). All but one standard agreed with the reference value within the stated uncertainty. No significant difference in performance was observed between standards made from synthetic air or whole air, and the accuracy of both types of standards was limited only by the ability to measure trace CH4 levels in the matrix gases used to produce the standards.

Final report on key comparison CCQM-K55.b (aldrin): An international comparison of mass fraction purity assignment of aldrin

Under the auspices of the Organic Analysis Working Group (OAWG) of the Comit? Consultatif pour la Quantit? de Mati?re (CCQM) a key comparison, CCQM K55.b, was coordinated by the Bureau International des Poids et Mesures (BIPM) in 2010/2011. Nineteen national measurement institutes and the BIPM participated. Participants were required to assign the mass fraction of aldrin present as the main component in the comparison sample for CCQM-K55.b which consisted of technical grade aldrin obtained from the National Measurement Institute Australia that had been subject to serial recrystallization and drying prior to sub-division into the units supplied for the comparison. Aldrin was selected to be representative of the performance of a laboratorys measurement capability for the purity assignment of organic compounds of medium structural complexity [molar mass range 300 Da to 500 Da] and low polarity (pKOW < ?2) for which related structure impurities can be quantified by capillary gas phase chromatography (GC). The key comparison reference value (KCRV) for the aldrin content of the material was 950.8 mg/g with a combined standard uncertainty of 0.85 mg/g. The KCRV was assigned by combination of KCRVs assigned by consensus from participant results for each orthogonal impurity class. The relative expanded uncertainties reported by laboratories having results consistent with the KCRV ranged from 0.3% to 0.6% using a mass balance approach and 0.5% to 1% using a qNMR method. The major analytical challenge posed by the material proved to be the detection and quantification of a significant amount of oligomeric organic material within the sample and most participants relying on a mass balance approach displayed a positive bias relative to the KCRV (overestimation of aldrin content) in excess of 10 mg/g due to not having adequate procedures in place to detect and quantify the non-volatile content?specifically the non-volatile organics content?of the comparison sample. There was in general excellent agreement between participants in the identification and the quantification of the total and individual related structure impurities, water content and the residual solvent content of the sample. The comparison demonstrated the utility of 1H NMR as an independent method for quantitative analysis of high purity compounds. In discussion of the participant results it was noted that while several had access to qNMR estimates for the aldrin content that were inconsistent with their mass balance determination they decided to accept the mass balance result and assumed a hidden bias in their NMR data. By contrast, laboratories that placed greater confidence in their qNMR result were able to resolve the discrepancy through additional studies that provided evidence of the presence of non-volatile organic impurity at the requisite level to bring their mass balance and qNMR estimates into agreement. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

International comparison CCQM-K82: methane in air at ambient level (1800 to 2200) nmol/mol

The CCQM-K82 comparison was designed to evaluate the degrees of equivalence of NMI capabilities for methane in air primary reference mixtures in the range (1800 to 2200) nmol/mol. The balance gas for the standards was either scrubbed dry real air or synthetic air. CH4 in air standards have been produced by a number of laboratories for many years, with more recent developments focused on standards at atmospheric measurement concentrations and aimed at obtaining agreement between independently produced standards. A comparison of the differences in primary gas standards for methane in air was previously performed in 2003 (CCQM-P41 Greenhouse gases. 1 and 2) with a standard deviation of results around the reference value of 30 nmol/mol and 10 nmol/mol for a more limited set of standards. This can be contrasted with the level of agreement required from field laboratories routinely measuring atmospheric methane levels, set by Data Quality Objectives (DQO) established by the World Meteorological Organization (WMO) to reflect the scientifically desirable level of compatibility for CH4 measurements at the global scale, currently set at 2 nmol/mol (1 sigma). The measurements of this key comparison took place from May 2012 to June 2012. Eight laboratories took part in this comparison coordinated by the BIPM and NIST. Key comparison reference values were calculated based on Cavity Ring Down Spectroscopy Measurements performed at the BIPM, combined with participants gravimetric values to identify a consistent set of standards. Regression analysis allowed predicted values for each standard to be calculated which acted as the KCRVs. In this comparison reported standard uncertainties by participants ranged from 0.50 nmol/mol to 2.4 nmol/mol and the uncertainties of individual KCRVs ranged from 0.68 nmol/mol to 0.71 nmol/mol. The standard deviation of the ensemble of standards about the KCRV value was 1.70 nmol/mol. This represents a greater than tenfold improvement in the level of compatibility of methane in air standards compared to that demonstrated in 2003. Further improvements in the compatibility of standards will require improved methods and uncertainties for the measurement of trace level methane in balance gases. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Final report on key comparison CCQM-K55.c (L-(+)-Valine): Characterization of organic substances for chemical purity

Under the auspices of the Organic Analysis Working Group (OAWG) of the Comit? Consultatif pour la Quantit? de Mati?re (CCQM) a key comparison, CCQM K55.c, was coordinated by the Bureau International des Poids et Mesures (BIPM) in 2012. Twenty National Measurement Institutes or Designated Institutes and the BIPM participated. Participants were required to assign the mass fraction of valine present as the main component in the comparison sample for CCQM-K55.c. The comparison samples were prepared from analytical grade L-valine purchased from a commercial supplier and used as provided without further treatment or purification. Valine was selected to be representative of the performance of a laboratorys measurement capability for the purity assignment of organic compounds of low structural complexity [molecular weight range 100?300] and high polarity (pKOW > ?2). The KCRV for the valine content of the material was 992.0 mg/g with a combined standard uncertainty of 0.3 mg/g. The key comparison reference value (KCRV) was assigned by combination of KCRVs assigned from participant results for each orthogonal impurity class. The relative expanded uncertainties reported by laboratories having results consistent with the KCRV ranged from 1 mg/g to 6 mg/g when using mass balance based approaches alone, 2 mg/g to 7 mg/g using quantitative 1H NMR (qNMR) based approaches and from 1 mg/g to 2.5 mg/g when a result obtained by a mass balance method was combined with a separate qNMR result. The material provided several analytical challenges. In addition to the need to identify and quantify various related amino acid impurities including leucine, isoleucine, alanine and ?-amino butyrate, care was required to select appropriate conditions for performing Karl Fischer titration assay for water content to avoid bias due to in situ formation of water by self-condensation under the assay conditions. It also proved to be a challenging compound for purity assignment by qNMR techniques. There was overall excellent agreement between participants in the identification and the quantification of the total and individual related structure impurities, water content, residual solvent and total non-volatile content of the sample. Appropriate technical justifications were developed to rationalise observed discrepancies in the limited cases where methodology differences led to inconsistent results. The comparison demonstrated that to perform a qNMR purity assignment the selection of appropriate parameters and an understanding of their potential influence on the assigned value is critical for reliable implementation of the method, particularly when one or more of the peaks to be quantified consist of complex multiplet signals. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

An international comparison of mass fraction purity assignment of theophylline: The Comité Consultatif pour la Quantité de Matière (CCQM) Pilot Study CCQM-P20.f (Digoxin)

Under the auspices of the Organic Analysis Working Group (OAWG) of the Comite Consultatif pour la Quantite de Matiere (CCQM) a laboratory comparison, CCQM-P20.f, was coordinated by the Bureau International des Poids et Mesures (BIPM) in 2007/2008. Nine national measurement institutes, four expert laboratories and the BIPM participated in the comparison. Participants were required to assign the mass fraction of digoxin present as the main component in the comparison sample (CCQM-P20.f) which consisted of digoxin material obtained from a commercial supplier stated to comply with USP requirements. In addition to assigning the mass fraction content of digoxin for the material, participants were requested, but not obliged, to provide mass fraction estimates for the minor components they identified in each sample. In contrast with the previous round of the CCQM-P20 series, in which the mass fraction content of theophylline in two comparison samples (CCQM-P20.e.1 and CCQM-P20.e.2) was determined, a wider range of results were reported for the mass fraction content of digoxin in the CCQM-P20.f comparison. A minority of participants did not appear to use conditions capable of fully resolving and/or quantifying the major related structure impurities present in the comparison sample. Among those that did achieve suitable separations, there was further variation in their reported quantifications of the individual and total related substance content which reflected in part the limited availability of reference standards for these materials and the resulting assumptions that had to be made regarding the structure and response factors relative to digoxin for each individual impurity. This was particularly relevant because of the span of molecular masses of the impurities present in the sample, which ranged from aglycones to glycones with tetrameric carbohydrate chains, relative to that of digoxin. A significant additional factor also contributed to the observed variation of results. Unlike the CCQM-P20.e samples, in which the major impurities were solely related structure organic compounds, the CCQM-P20.f study material contained significant levels of residual organic solvents (ethanol, dichloromethane and to a lesser extent toluene). The majority of participants failed to detect and allow for the presence of this class of impurity, introducing a bias towards overestimation of digoxin content in most of the individual results. However, the uncertainty budgets produced by several participants were sufficiently conservative such that their reported results were nevertheless consistent with the reference value for digoxin content assigned using a consensus mass balance approach. The results of the comparison reinforce the conclusion from previous rounds of the CCQM-P20 study that care in developing and validating the suitability of the chromatographic separation method used to resolve the main component from the related structure impurities present is essential to obtaining reliable, comparable results when using the mass balance approach to estimate purity. This specific comparison has demonstrated that, in addition to developing an appropriate chromatographic separation, it is also important to use complementary techniques capable of detecting all potential orthogonal classes of impurities if it is desired to demonstrate a general capability to assign purity with a small (<0.2% relative) standard uncertainty. Main text. To reach the main text of this paper, click on Final Report. The final report has been peer-reviewed and approved for publication by the CCQM OAWG.

Normal phase-liquid chromatography-tandem mass spectrometry with atmospheric pressure photoionization for the purity assessment of 17β-estradiol

AbstractA normal phase-liquid chromatography-hybrid tandem mass spectrometry (NP-LC-MS/MS) method utilizing atmospheric pressure photoionization (APPI) without dopant has been developed and implemented for the simultaneous determination of several estrogenic steroid hormones. The combination of both NP-LC and APPI-MS/MS tolerates the use of solvents that have the advantages of being self-doping for APPI and, at the same time, inhibit the in situ formation of estrogen dimers as frequently observed for conventional reversed phase (RP)-LC methods. The NP-LC-APPI-MS/MS method has been validated in-house, and its performance characteristics (linearity, repeatability, limits of detection, etc.) were assessed for use in the quantification of estrogens. Moreover, the method was used to characterize and determine the inherent related structure impurities in batches of β-estradiol, required for the establishment of reference measurement systems for clinical chemistry and laboratory medicine, which served as candidate reference material for an organic purity assessment interlaboratory study (CCQM-K55.a) organized by the International Bureau of Weights and Measures (BIPM) Chemistry Department and carried out within the framework of the Organic Analysis Working Group (OAWG) of the Consultative Committee for Amount of Substance—Metrology in Chemistry (CCQM). Graphical abstractFull-scale and enlarged overlaid SRM chromatograms of three transitions for ten estrogenic steroid hormones for the impurity quantification of a βE2 material