710 es spectrometer

Boron and cerium were determined using inductively coupled plasma optical emission spectroscopy (ICP-OES) with a Varian 710-ES spectrometer (Varian, Mulgrave, Victoria, Australia). The following parameters were used: radio frequency (RF) power 1.0 kW, plasma flow 15 dm³/min, auxiliary flow 1.5 dm³/min, nebuliser pressure 200 kPa, pump rate 15 rpm and emission lines: λ = 208.956, λ = 249.678 and λ = 249.772 nm (for boron) and λ = 407.347, λ = 407.570, λ = 418.659 and λ = 446.021 nm (for cerium). Calibration curves were prepared using standard solutions in the ranges of 0.1–1.5 mg/dm³ and 0.1–5 mg/dm³ for boron and cerium, respectively. The changes in the chitosan structure were determined by a Fourier-transform infrared (FTIR) Spectrum Two spectrometer (Perkin Elmer, Waltham, MA, USA). The scanning electron microscope (SEM) micrographs of the chitosan bio-composite were produced using a Phenom ProX SEM (Phenom-World Bv, Eindhoven, The Netherlands). The phase compositions of the chitosan-based samples were determined using a Seifert 3003TT powder X-ray diffractometer with a Cu X-ray tube: kλ₁ = 1.540598 Å, kλ₂ = 1.544426 Å and kβ = 1.39225 Å (Seifert, Ahrensburg, Germany). Powder samples were analysed three times, and the XRD patterns presented in the manuscript were an average of the scans between 10 and 90° 2Theta.

The tests were carried out in an incubator at 37 °C in 20 mL
of Ringer solution on a shaker at 60 rpm for 12 weeks. The release
of Ti, Ca, and P was determined with inductively coupled plasma-optical
emission spectroscopy (ICP-OES) with a Varian 710-ES spectrometer
(Varian Inc., Palo Alto, CA, USA) using parameters as described.^{17 (link)}

To quantitate the iron content in the collected lesion and muscle tissues, samples were dried, weighed, and digested with 69% nitric acid (TraceSELECT™, Fluka, St. Louis, MO, USA) at room temperature. The iron content in each sample was determined through inductively coupled plasma optical emission spectrometry (ICP-OES) utilizing a 710-ES spectrometer (Varian, Palo Alto, CA, USA). Blank nitric acid and calibration samples containing a predetermined iron concentration were prepared using TraceCERT^® Iron (Fe) Standard for ICP (1000 mg/L Fe in nitric acid, Sigma-Aldrich, St. Louis, MO, USA) and analyzed alongside the test tissue samples. Measurements were carried out in triplicate, and the resulting data were normalized to the dry tissue weight and presented as mean ± SD.

The anodized samples were placed separately in Falcon tubes, each with 20 mL of Ringer solution. The samples were shaken at 60 rpm and incubated at 37 °C (incubator POL-EKO, Wodzisław Śląski, Poland) for up to 6 weeks. After each 2-week period, the content of niobium, phosphorus, zirconium, calcium, and silver released into the Ringer solution was determined using inductively coupled plasma atomic emission spectrometry (ICP-AES). A Varian 710-ES spectrometer (Santa Clara, USA). equipped with a OneNeb nebulizer was utilized. The parameters were as follows: RF power 1.0 kW, plasma flow 15 L min⁻¹, auxiliary flow 1.5 L min⁻¹, nebulizer pressure 210 kPa, pump rate 15 rpm; emission lines of Nb:λ = 295.088 and 294.154 nm, P:λ = 213.618, Zr:λ = 343.823 and 327.307 nm, Ca:λ = 317.933 and 422.673 nm, and Ag:λ = 328.068 and 338.289 nm. The calibration curve method was applied. The calibration curves were prepared using a matrix (Ringer solution) at the same concentration as that of the samples. Single element standard solutions of 1 and 10 mg mL⁻¹ (zirconium) supplied by Merck Millipore (Darmstadt, Germany) were used. Deionized water was prepared using a Millipore Elix 10 system (Darmstadt, Germany). The obtained results are the average of concentrations obtained for all used analytical lines with the standard deviation not exceeding 1.5%.

To quantitate gadolinium content in the collected lesions and muscle tissues, samples were dried, weighed, and digested with 69% nitric acid (TraceSELECT™, Fluka, USA). Samples were then diluted further to 4% nitric acid and filtered to obtain a solution with no visible debris. Gadolinium content was determined for each sample by inductively coupled plasma optical emission spectrometry (ICP-OES) using a 710-ES spectrometer (Varian, Palo Alto, CA). For all ICP-OES measurements, blank nitric acid, and samples with known Gd concentration as calibration were prepared and tested concurrently with test tissue samples. All measurements were carried out in triplicates, and the data were normalized to the dry tissue weight and reported as mean ± SD.