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 results of an absolute silicon molar mass determination of two independent sets of samples from the highly 28Si-enriched crystal (AVO28) produced by the International Avogadro Coordination are presented and compared with results published by the Physikalisch-Technische Bundesanstalt (PTB, Germany), the National Research Council (NRC, Canada) and the National Metrology Institute of Japan (NMIJ, Japan). This study developed and describes significant changes to the published protocols for producing absolute silicon isotope ratios. The measurements were made at very high resolution on a multi-collector inductively coupled plasma mass spectrometer using tetramethylammonium hydroxide (TMAH) to dissolve and dilute all samples. The various changes in the measurement protocol and the use of TMAH resulted in significant improvements to the silicon isotope ratio precision over previously reported measurements and in particular, the robustness of the 29Si/30Si ratio of the AVO28 material. These new results suggest that a limited isotopic variability is present in the AVO28 material. The presence of this variability is at present singular and therefore its significance is not well understood. Fortunately, its magnitude is small enough so as to have an insignificant effect on the overall uncertainty of an Avogadro constant derived from the average molar mass of all four AVO28 silicon samples measured in this study. The NIST results confirm the AVO28 molar mass values reported by PTB and NMIJ and confirm that the virtual element–isotope dilution mass spectrometry approach to calibrated absolute isotope ratio measurements developed by PTB is capable of very high precision as well as accuracy. The Avogadro constant NA and derived Planck constant h based on these measurements, together with their associated standard uncertainties, are 6.02214076(19) × 1023 mol−1 and 6.62607017(21) × 10−34 Js, respectively.

Understanding the translocation of nanoparticles (NPs) into plants is challenging because qualitative and quantitative methods are still being developed and the comparability of results among different methods is unclear. In this study, uptake of titanium dioxide NPs and larger bulk particles (BPs) at 5 mg/L and 50 mg/L concentrations in rice plant (Oryza sativa L.) tissues was evaluated using three orthogonal techniques: electron microscopy, single-particle inductively coupled plasma mass spectroscopy (spICP-MS) with two different plant digestion approaches, and total elemental analysis using ICP optical emission spectroscopy. In agreement with electron microscopy results, total elemental analysis of plants exposed to TiO2 NPs and BPs at 5 mg/L and 50 mg/L concentrations revealed that TiO2 NPs penetrated into the plant root and resulted in Ti accumulation in above ground tissues at a higher level compared to BPs. spICP-MS analyses revealed that the size distributions of internalized particles differed between the NPs and BPs with the NPs showing a distribution with smaller particles. Acid digestion resulted in higher particle numbers and the detection of a broader range of particle sizes than the enzymatic digestion approach, highlighting the need for development of robust plant digestion procedures for NP analysis. Overall, there was agreement among the three techniques regarding NP and BP penetration into rice plant roots and spICP-MS showed its unique contribution to provide size distribution information.

The total amount of DNA in a preparation extracted from tissues can be measured in several ways, each method offering advantages and disadvantages. For the sake of accuracy in quantitation, it is of interest to compare these methodologies and determine if good correlation can be achieved between them. Different answers can also be clues to the physical state of the DNA. In this study, we investigated the lack of correlation between ultraviolet (UV) absorbance and fluorescent (PicoGreen) measurements of the concentration of DNAs isolated from plant tissues. We found that quantitation based on the absorbance-based method correlated with quantitation based on phosphorus content, while the PicoGreen-based method did not. We also found evidence of the production of single-stranded DNA under conditions where the DNA was not fragmented into small pieces. The PicoGreen fluorescent signal was dependent on DNA fragment size but only if the DNA was in pure water, while DNA in buffer was much less sensitive. Finally, we document the high sensitivity of the PicoGreen assays to the detergent known as CTAB (cetyldimethylethylammonium bromide). The CTAB-based method is highly popular for low-cost DNA extraction with many published variations for plant and other tissues. The removal of residual CTAB is important for accurate quantitation of DNA using PicoGreen.