Liquid chromatographic analysis of vitamin B6 in reconstituted infant formula: collaborative study

A liquid chromatographic (LC) method was validated for the determination of total vitamin B6 in infant formula. Total vitamin B6 was quantified by converting the phosphorylated and free vitamers into pyridoxine. Pyridoxine was determined by ion pair reversed-phase LC with fluorescence detection. The method was subjected to an AOAC collaborative study involving a factory-manufactured, milk- and soy-based infant formula. Each was spiked at 3 concentrations in the range of 0-1 microg/g and sent as blind duplicate to participant laboratories. Nine laboratories returned valid data which were statistically analyzed for outliers and precision parameters. The repeatability relative standard deviation (RSD(r)) ranges were 2.0-4.0 and 3.5-5.9% for fortified milk- and soy-based formulas, respectively. The reproducibility relative standard deviation (RSD(R)) ranges were 8.2-8.4 and 6.7-11.2% for fortified milk- and soy-based formulas, respectively. HORRAT values ranged from 0.42 to 0.53, indicating that the precision of the method is acceptable. The mean RSD(r):RSD(R) values were 0.60 and 0.55 for milk- and soy-based formulas, respectively. As expected, RSDs for the unfortified samples were higher, but their HORRAT values (0.81 and 2.06) helped define a realistic limit of quantitation as 0.05 microg/g. Recovery data were quantitative and varied between 81.4 and 98.0% (mean = 89.8%) for each of 6 spiked materials.     

FREQUENCY OF ULTRAVIOLET RADIATION-INDUCED MUTATION AT THE hprt LOCUS IN REPAIR-PROFICIENT MURINE FIBROBLASTS TRANSFECTED WITH THE denV GENE OF BACTERIOPHAGE T4

Abstract— Thc frequency of spontaneous and ultraviolet radiation (UVR)-induced mutation at the hprt locus was determined in control and denV-transfected, repair-proficient murine fibroblasts. Control cells removed an average of 25% of pyrimidine dimers induced by exposure to 150 J/m²UVR from an FS40 sunlamp within 24 h; under the same conditions of induction and repair, denV-transfected cells removed an average of 71% of pyrimidine dimers. Control cells were somewhat more resistant than denV-transfected cells to killing by UVR. The average frequency of spontancous mutation at the hprt locus for control and denV-transfected cells was 3 and 15 6-thioguanine (6-TG)-resistant colonies per 10⁶ surviving cells, respectively; there was no statistically significant difference between control and dcnV-transfected cells. However, after exposure to 75 or 150 J/m² UVR, denV-transfected cells had a significantly lower frequency of mutation to 6-TG resistance. After exposure to a fluence of 75 J/m², the average frequency of UVR-induced mutation at the hprt locus was 166 mutant colonies per lo^h surviving cells for control cells and 92 mutant colonies for denV-transfectcd cells; after 150 J/m², control cells had 205 6-TG-resistant colonies per 10⁶ cells, while dmV-transfected cclls had 61 mutant colonies. These results demonstrate that UVR-induced pyrimidine dimers are mutagenic photoproducts in mammalian cells.     

Insulator–metal transition in Nd0.8Na0.2Mn(1−x)CoxO3 perovskites

The electric and magnetic properties of the perovskites Nd_0.8Na_0.2Mn_(1−x)Co_xO₃ (0x0.2) prepared by the usual ceramic procedure were investigated. The insulator-to-metal-like (IM) transition, closely related to a ferromagnetic arrangement, was revealed for the composition of x=0.04 and a similar tendency was detected for x=0. The insulating behavior persists down to low temperatures for higher contents of cobalt ions in spite of the transition to the bulk ferromagnetism. The properties are interpreted in terms of the steric distortion, tilting of the Mn(Co)O₆ octahedra and the double-exchange interactions of the type Mn³⁺–O²⁻–Mn⁴⁺and Mn^3.5+δ–O²⁻–Co²⁺, respectively. Presence of antiferromagnetic domains in the ferromagnetic matrix for the most of cobalt-substituted samples is supposed.     

Solution and solid-state structure of the anion [Ag(2)[closo-CB(11)H(12)](4)](2-)

Addition of the carbene 1,3-dimesitylimidazol-2-ylidene (IMes) to a toluene solution of Ag[closo-CB(11)H(12)] results in the formation of the complex [(IMes)(2)Ag](2)[Ag(2)[closo-CB(11)H(12)](4)], the anionic component of which contains two silver(I) centers bridged by two carboranes in addition to one terminally bound carborane on each metal, in the solid-state. Comparison of the observed (11)B[(1)H] NMR chemical shifts of [(IMes)(2)Ag](2)[Ag(2)[closo-CB(11)H(12)](4)] or Ag[closo-CB(11)H(12)] with [NBu(4)][closo-CB(11)H(12)] in CD(2)Cl(2) demonstrates that the silver ion interacts significantly with the cage in solution. Theoretical investigations using the ab initio/GIAO/NMR method of [closo-CB(11)H(12)](-) and Na[closo-CB(11)H(12)] as model geometries for the silver salts support experimental evidence for these Ag...[BH] interactions in solution.     

The two-photon excitation cross section of 6MAP, a fluorescent adenine analogue

6MAP is a fluorescent analogue of adenine that undergoes Watson-Crick base pairing and base stacking in double-stranded DNA. The one-photon absorption spectrum of 6MAP is characterized by a maximum around 330 nm with moderate quantum yield fluorescence centered at about 420 nm. To take advantage of this probe for confocal and single-molecule microscopy, it would be advantageous to be able to excite the analogue via two photons. We report the first determination of the two-photon excitation cross section and spectrum for 6MAP from 614 to 700 nm. The power dependence of the fluorescence indicates that emission results from the absorption of two photons. The one-photon and two-photon emission line shapes are identical within experimental error. A study of the concentration dependence of the fluorescence yield for one-photon excitation shows no measurable quenching up to about 5 microM. The maximum in the two-photon excitation spectrum gives a two-photon cross section, delta(TPE), of 3.4 +/- 0.1 Goeppert-Mayer (G.M.) at 659 nm, which correlates well with the one-photon absorption maximum. This compares quite favorably with cross sections of various naturally fluorescent biological molecules such as flavins and nicotiamide. In addition, we have also obtained the two-photon-induced fluorescence emission spectrum of quinine sulfate. It is approximately the same as that for one-photon excitation, suggesting that two-photon excitation of quinine sulfate may be used for calibration purposes.     

Modeling features of the non-heme diiron cores in O2-activating enzymes through the synthesis, characterization, and oxidation of 1,8-naphthyridine-based complexes

Multidentate naphthyridine-based ligands were used to prepare a series of diiron(II) complexes. The compound [Fe(2)(BPMAN)(mu-O(2)CPh)(2)](OTf)(2) (1), where BPMAN = 2,7-bis[bis(2-pyridylmethyl)aminomethyl]-1,8-naphthyridine, exhibits two reversible oxidation waves with E(1/2) values at +310 and +733 mV vs Cp(2)Fe(+)/Cp(2)Fe, as revealed by cyclic voltammetry. Reaction with O(2) or H(2)O(2) affords a product with optical and M?ssbauer properties that are characteristic of a (mu-oxo)diiron(III) species. The complexes [Fe(2)(BPMAN)(mu-OH)(mu-O(2)CAr(Tol))](OTf)(2) (2) and [Fe(2)(BPMAN)(mu-OMe)(mu-O(2)CAr(Tol))](OTf)(2) (3) were synthesized, where Ar(Tol)CO(2)(-) is the sterically hindered ligand 2,6-di(p-tolyl)benzoate. Compound 2 has a reversible redox wave at +11 mV, and both 2 and 3 react with O(2), via a mixed-valent Fe(II)Fe(III) intermediate, to give final products that are also consistent with (mu-oxo)diiron(III) species. The paddle-wheel compound [Fe(2)(BBAN)(mu-O(2)CAr(Tol))(3)](OTf) (4), where BBAN = 2,7-bis(N,N-dibenzylaminomethyl)-1,8-naphthyridine, reacts with dioxygen to yield benzaldehyde via oxidative N-dealkylation of a benzyl group on BBAN, an internal substrate. In the presence of bis(4-methylbenzyl)amine, the reaction also produces p-tolualdehyde, revealing oxidation of an external substrate. A structurally related compound, [Fe(2)(BEAN)(mu-O(2)CAr(Tol))(3)](OTf) (5), where BEAN = 2,7-bis(N,N-diethylaminomethyl)-1,8-naphthyridine, does not undergo N-dealkylation, nor does it facilitate the oxidation of bis(4-methylbenzyl)amine. The contrast in reactivity of 4 and 5 is attributed to a difference in accessibility of the substrate to the diiron centers of the two compounds. The M?ssbauer spectroscopic properties of the diiron(II) complexes were also investigated.     

On the lower Markov spectrum

C. Hightower found two infinite sequences of gaps in the Markov spectrum, ( _n , _n ) and ( _n , _n ) with _n and _n both Markov elements, converging to . This paper exhibits Markov elements _n ^* and _n ^* such that, for alln 1, ( _n ^* , _n ) and ( _{n n} ^* ) are gaps in the Markov spectrum. Other results include showing that, for alln 1, _n is completely isolated, while the other endpoints of the gaps are limit points in the Markov spectrum.     

The estimation and use of measurement uncertainty for a drug substance test procedure validated according to USP <1225>

The vision for metrology at the United States Pharmacopeia (USP) is for international recognition, harmonization, and official acceptance of all USP reference standards that are based on sound, scientific, metrological principles, such as measurement uncertainty (MU). Pharmaceutical testing laboratories will need to estimate the MU for their test procedures. This paper demonstrates how to estimate MU for a test procedure validated following the requirements in USP General Chapter <1225>, Validation of Compendial Procedures. The test procedure determines the assay and impurities for a pharmaceutical drug substance. Pharmaceutical manufacturing companies are required to test the drug substance used in their drug products. In this example, a new test procedure was developed and validated following the requirements in USP <1225>. The MU was estimated using the data from the test procedure validation. The MU estimate determined that the test procedure was fit for use with one determination, avoiding the expense of duplicate determinations. The MU estimate was used to set up decision rules for comparing test data to the assay and impurity specifications. The information from the MU estimate was used to monitor the routine use of the test procedure using control charts that assess accuracy and precision of each run based upon actual test procedure performance capability.