Method for the fixation of optrodes in near-infra-red spectrophotometry.
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One of the major threats to patients undergoing maintenance hemodialysis is an increased susceptibility to infections caused by microorganisms, among which Yersinia or Listeria are frequently recovered. In patients receiving hemodialysis, iron overload owing to multiple transfusions plays an important role in the mechanisms underlying the susceptibility to bacterial infections, partially mediated by impaired neutrophil function. In order to assess the true role of iron at the cellular level, an AAS method was developed to measure the iron content of granulocytes. Iron levels in the granulocytes were determined in an apparently healthy population and in a population of iron-overloaded renal hemodialysis patients. Granulocytes were isolated by a method modified from Böyum. The analyses were performed using pyrocoated graphite tubes, and in one of the char steps, oxygen was used to facilitate ashing. The mean iron level found in the granulocytes from apparently normal persons was 4.07 fg/cell (n = 17) with a CV of 44%; the mean value for the dialysis patient group was 4.59 fg/cell (n = 8) with a CV of 37%. There was no significant difference between the two groups, p = 0.70.
The acidic reduction of Hg using a continuous-flow analytical system was evaluated. With 25% SnCl2 as the reductant, characteristic concentrations (sensitivities) of 0.44 microgram/L (open cell) and 0.29 microgram/L (flow-through cell) were obtained using inorganic Hg2+ standards in 1.5% HCl. When CH3Hg+ standards were used, absorption signals were an order of magnitude lower, indicating that Sn(II) is incapable of producing Hg degree from organic Hg in this acidic reduction system. Addition of CdCl2 to the SnCl2 reductant, as suggested by Magos (1) for the reduction of organomercurials under alkaline conditions, was without beneficial effect. Similarly, combining Sn with another reducing agent (hydroxylamine hydrochloride), or a strong alkaline solution (40% NaOH), in the reaction coil of the flow-through system did not significantly enhance the Hg absorption signal for either inorganic or organic Hg. Recovery of Hg from spiked liver homogenates digested at 70-80 degrees C using a HNO3/H2SO4/HCl procedure and stabilized with 0.5 mM K2Cr2O7 was > 85% using either inorganic Hg2+ or CH3Hg+, indicating that this digestion procedure successfully breaks the C-Hg bond to form readily reducible Hg species. Using L-cysteine to stabilize standards of inorganic Hg2+ in HCl caused significant depressions of the Hg absorption signal at L-cysteine concentrations > 0.001% (approximately 0.5 mM); 0.1% L-cysteine caused total suppression of the Hg signal. These results indicate that: (1) acidic reduction of Hg by Sn in this continuous-flow system requires breakdown of organomercurials prior to analysis; (2) tissue digestion using HNO3/H2SO4/HCl followed by the addition of K2Cr2O7 to stabilize Hg2+ achieves this breakdown and allows good recovery of total Hg; and (3) use of L-cysteine to complex and prevent losses of Hg should be avoided in systems using acidic reduction of Hg. Concentrations of endogenous tissue sulfhydryls are generally lower than those associated with depressed absorbance signals during the acidic reduction of Hg.
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A new highly sensitive and selective chromogenic reagent, p-carboxyphenylfluorone (p-CPF), was studied for spectrophotometric determination of trace molybdenum. In 0.36 mol L(-1)phosphoric acid medium, p-CPF reacts with molybdenum(VI) to form a 1:3 red complex, which has a sensitive absorption peak at 531 nm. Under optimal conditions, the reaction of molybdenum(VI) with p-CPF completed rapidly and absorbance remains almost constant for at least 24 h. Molybdenum(VI) obeyed Beer's law in the range 0-2.0 microg mL(-1); the apparent molar absorption coefficient, Sandell's sensitivity and the limit of detection were found to be 1.03 x 10(5) L mol(-1) cm(-1), 1.028 ng cm(-2)and 0.73 ng mL(-1) respectively; the effect of various foreign ions were examined in detail. It was found that most coexisting ions can be tolerated in considerable amounts, especially 800 mg of Mn(II), 200 mg of Mg(II), Fe(II), Co(III), Ni(II) and Cd(II), 50 mg of Ca(II) and Al(III), 25 mg of Cu(II) and Fe(III), 10 mg of Hg (II), La(III), Bi(III), Pb(II) and Zn(II) don't interfere with the determination of molybdenum(VI). The proposed method is very simple, sensitive and selective, it has been applied to determine molybdenum in vegetable and food samples with a very high precision and accuracy. Moreover, the synthesis of the reagent and the conditions of the colour reaction were also studied in detail.
The interaction between azur A (AA) and hyaluronic acid (HA) at AA concentrations from 3.430 x 10(-5) to 8.575 x 10(-5) M and sodium chloride concentrations from 0 to 0.01 M was investigated spectrophotometrically at 620 nm at temperatures from 0 to 50 degrees C. AA was shown to be a useful spectroscopic probe for detecting carboxyl groups in HA macromolecules. The interaction between AA and HA was temperature sensitive and little AA-HA interaction was observed at temperatures higher than 30 degrees C. The interaction of HA with AA was seen to be electrostatic in nature. The maximum binding number decreased with decreasing NaCl concentration, and the absorbance sensitivity decreased with increasing NaCl concentration in aqueous solution. Self-interference from the AA in the AA-HA interaction caused an overestimate of the molar mass of hyaluronic acid. An improved method was proposed to estimate the molar mass of HA, and a molar mass of 1.219 x 10(6) Da was obtained with this improved method for HA.
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