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This paper concerns an international research project aimed at determining the Avogadro constant by counting the atoms in an isotopically enriched silicon crystal. The counting procedure was based on the measurement of the molar volume and the volume of an atom in two 1 kg crystal spheres. The novelty was the use of isotope dilution mass spectrometry as a new and very accurate method for the determination of the molar mass of enriched silicon. Because of an unexpected metallic contamination of the sphere surfaces, the relative measurement uncertainty, 3 × 10−8 NA, is larger by a factor 1.5 than that targeted. The measured value of the Avogadro constant, NA = 6.022 140 82(18) × 1023 mol−1, is the most accurate input datum for the kilogram redefinition and differs by 16 × 10−8 NA from the CODATA 2006 adjusted value. This value is midway between the NIST and NPL watt-balance values.
New results are reported from an ongoing international research effort to accurately determine the Avogadro constant by counting the atoms in an isotopically enriched silicon crystal. The surfaces of two 28 Si-enriched spheres were decontaminated and reworked in order to produce an outer surface without metal contamination and improved sphericity. New measurements were then made on these two reconditioned spheres using improved methods and apparatuses. When combined with other recently refined parameter measurements, the Avogadro constant derived from these new results has a value of N A = 6.022 140 76(12) × 10 23 mol -1 . The X-ray crystal density method has thus achieved the target relative standard uncertainty of 2.0 × 10 -8 necessary for the realization of the definition of the new kilogram.
The FDA's process analytical technology initiative encourages drug manufacturers to apply innovative ideas to better understand their processes. There are many challenges to applying these techniques to monitor mammalian cell culture bioreactors for biologics manufacturing. These include the ability to monitor multiple components in complex medium formulations non-invasively and in-line.We report results that demonstrate, for the first time, the technical feasibility of the in-line application of Raman spectroscopy for monitoring a mammalian cell culture bioreactor. A Raman probe was used for the simultaneous prediction of culture parameters including glutamine, glutamate, glucose, lactate, ammonium, viable cell density, and total cell density.
The Avogadro constant links the atomic and the macroscopic properties of matter. Since the molar Planck constant is well known via the measurement of the Rydberg constant, it is also closely related to the Planck constant. In addition, its accurate determination is of paramount importance for a definition of the kilogram in terms of a fundamental constant. We describe a new approach for its determination by counting the atoms in 1 kg single-crystal spheres, which are highly enriched with the 28Si isotope. It enabled isotope dilution mass spectroscopy to determine the molar mass of the silicon crystal with unprecedented accuracy. The value obtained, NA = 6.022,140,78(18) × 10(23) mol(-1), is the most accurate input datum for a new definition of the kilogram.
Glycosylation patterns and specific expression rates of the recombinant protein mouse placental lactogen-I (mPL-I) by Chinese hamster ovary (CHO) cells varied significantly over the extracellular pH (pHe) range of 6.1 to 8.7. The maximum specific mPL-I expression rates occurred between pHe 7.6 and 8.0. The pHe effect on protein expression was confirmed using a different CHO cell expressing the unglycosylated recombinant protein mouse placental lactogen-II (mPL-II). Decreases in the extent of glycosylation of mPL-I were observed at low (below 6.9) and high (above 8.2) pHe values. The pHe dependent variations in mPL-I accumulation in the supernatant as well as in glycosylation patterns were not the result of enzymatic degradation in the culture medium.
When the kilogram is redefined in terms of the fixed numerical value of the Planck constant h, the x-ray-crystal-density (XRCD) method, among others, is used for realizing the redefined kilogram. The XRCD method has been used for the determination of the Avogadro constant N A by counting the number of atoms in a 28 Si-enriched crystal, contributing to a substantial reduction of uncertainty in the values of N A and h to 2 parts in 10 8 . This method can be therefore used reversely for the mass determination of a 1 kg sphere prepared from the crystal. This is realized by SI-traceable measurements of its lattice parameter, isotopic composition, volume, and surface properties. Details of the corresponding measurements are provided, as well as the concept of the XRCD method, isotope enrichment, crystal production, sphere manufacturing, and evaluation of impurities and self-point defects in the crystal, together with mass comparison with respect to the silicon sphere for disseminating mass standards.
A new single crystal from isotopically enriched silicon was used to determine the Avogadro constant N A by the x-ray-crystal density method. The new crystal, named Si28-23Pr11, has a higher enrichment than the former 'AVO28' crystal allowing a smaller uncertainty of the molar mass determination. Again, two 1 kg spheres were manufactured from this crystal. The crystal and the spheres were measured with improved and new methods. One sphere, Si28kg01a, was measured at NMIJ and PTB with very consistent results. The other sphere, Si28kg01b, was measured only at PTB and yielded nearly the same Avogadro constant value. The mean result for both 1 kg spheres is N A = 6.022 140 526(70) × 10 23 mol −1 with a relative standard uncertainty of 1.2 × 10 −8 . This value deviates from the Avogadro value published in 2015 for the AVO28 crystal by about 3.9(2.1) × 10 −8 . Possible reasons for this difference are discussed and additional measurements are proposed.
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