Icp ms

TiO₂NPs were purchased from Sigma-Aldrich (particle size <25 nm, 637254, St. Louis, MO, USA). We quantified the morphology and size of TiO₂NPs using transmission electron microscopy (JEM-1210, JEOL, Tokyo, Japan) and scanning electron microscopy (Zeiss EVO-MA10; Carl Zeiss Meditec AG, Jena, Germany) at accelerating voltages of 200 kV and 15 kV, respectively. The specific surface area of TiO₂NPs was measured by nitrogen absorption methods based on the multipoint BET method (ASAP2020; Micromeritics, Norcross, GA, USA). The hydrodynamic size and zeta potential of TiO₂NPs were determined by ELS-8000 (Otsuka Electronic, Tokyo, Japan). The purity of TiO₂NPs used in the experiment was determined by energy-dispersive X-ray spectroscopy (Rayny EDX-700, Shimadzu, Kyoto, Japan). The endotoxin levels in thee TiO₂NP suspension were determined using a Pierce LAL Chromogenic Endotoxin Quantitation Kit (Thermo Fisher Scientific, Waltham, MA, USA). After completion of treatment procedures, lung tissue was harvested, weighed, and digested overnight with concentrated nitric acid, and the resultant samples were analyzed for elemental TiO₂NPs using ICP-MS (Perkin Elmer, Waltham, MA, USA).

The concentrations of trace elements were measured with ICP MS (Perkin Elmer Germany) according to the methods (no. 161) described by NMKL [26 ] in Food Control Laboratory in Izmir (Turkey). The concentrations obtained from the analyses were compared with the same tissue (muscle) and species (crayfish) in MFAL and EC. The detectable limits of elements were 0.003, 0.001, 0.05, 0.01, 0.01, and 0.01 mg kg⁻¹ for As, Hg, Zn, Cd, Cu, and Pb, respectively.

For trace element detection, 5 ml of fasting venous blood sample was collected and placed in a vacuum EDTA-K₂ anticoagulant blood collection vessel. After centrifugation at 3000 rpm for 10 min, the red blood cell fraction and plasma were removed, and the serum was transferred to a deionized cryotube and stored in a refrigerator at -20 °C until element analysis. The serum concentrations of Cu and Zn were evaluated using inductively coupled plasma spectrometry (ICP–MS, PerkinElmer Life and Analytical Sciences, Inc., CT, USA). For male participants with levels less than 0.70 mg/L and more than 1.40 mg/L and for female participants with levels less than 0.80 mg/L and more than 1.50 mg/L, the cut-offs were used to categorize hypo- and hypercupremia, respectively. The normal range of serum Zn levels was 0.76 mg/L to 1.50 mg/L. SCZR was evaluated as an alternative biomarker for assessing clinical outcomes [16 (link)].

Data on demographic characteristics, personal medical history, and family history of thyroid diseases were collected by interview during the survey. Life style risk factors (i.e., smoking status) were based on self-reporting. Serum TSH, FT4, and TPOAb concentrations were measured by an electrochemiluminescence immunoassay. Serum TSH was measured using an E-TSH kit (Roche Diagnostics, Mannheim, Germany). Serum FT4 was measured by E-Free T4 kit (Roche Diagnostics, Mannheim, Germany). Serum TPOAb was measured by E-Anti-TPO kit (Roche Diagnostics, Mannheim, Germany). The results of TSH, FT4, and TPOAb met the specifications regarding accuracy, general chemistry, special immunology, and ligand of the quality control and quality assurance program of the College of American Pathologists. UI was measured using an inductively coupled plasma mass spectrometry device (ICP-MS; Perkin Elmer ICP-MS, Waltham, MA, USA). The laboratory that measured UI is enrolled in the “Ensuring the Quality of Urinary Iodine Procedures (EQUIP)” quality assurance program run by the Centers for Disease Control of the United States of America [17 ].

The degradation behavior of (Ca,Sr)SO₄ specimen was evaluated in a phosphate buffered saline (PBS, Gibco Co. USA) solution. The composition of the solution, as reported by the manufacturer, was potassium chloride (KCl, 200 mg/L), potassium phosphate monobasic (KH₂PO₄, 200 mg/L), sodium chloride (NaCl, 8000 mg/L) and sodium phosphate dibasic (Na₂HPO₄^.7H₂O, 2160 mg/L). The specimen weight to the solution volume was kept at a constant ratio of 1 mg: 10 ml. The specimen and solution were shaken within a test tube to simulate the dynamic situation in vivo. The solution was refreshed every day. After each day, the remain of disc specimen was collected from the solution, then dried in an oven at 100 °C for 1 h. The weight of the specimen was measured. The rate of degradation was expressed in terms of weight loss on a daily basis. The concentration of Ca and Sr ions in the collected solution was measured with an inductively coupled plasma-mass spectroscopy (ICP-MS, Perkin Elmer Co., USA). The [Ca²⁺] and [Sr²⁺] in the fresh PBS were measured with the same technique. The pH value of the solution was also monitored.

The dried right rat femoral bones of both sham and OVX animals were weighted and the weight was expressed in grams. After weighing, the femoral bones were subjected to ashing in a muffle furnace (Lenton thermal designs, Parsons Lane, Hope, Hope Valley, S33 6RB, UK) for 16 hr at 700 °C. Next, the ashed bones were weighed and the weights were expressed as mg/100 mg net bone weight. Samples were then digested in 70% nitric acid (5 ml per each 100 mg ash) and the acidic mixtures were incubated shaking in a water bath (37 °C) for an overnight. Samples were then diluted with deionized distilled water (1:9 ratio)^{16 , 53 (link)}. On the other hand, 0.1 ml of serum and urine samples were directly digested with 0.9 ml 70% nitric acid, then, diluted with deionized distilled water (1:9 ratio). The diluted ash, serum and urine samples were used to quantify the concentrations of calcium, inorganic phosphorus and magnesium (Ca²⁺, P and Mg²⁺) by an inductive coupled plasma mass spectroscopy (ICP-MS) (PerkinElmer Life and Analytical Sciences 710 Bridgeport Avenue Shelton, CT, USA). Minerals concentrations in the femoral bones were expressed as mg mineral per 100 mg ash, while minerals concentrations in serum and urine were expressed as mmol/L. Urine results were normalized using creatinine concentration (mmol/L).

Cells or lysed tissue were washed twice and digested with a 3:1 ratio of trace element grade nitric acid:HCl for 20 min at 200 °C. Iron concentrations were analyzed using ICP–MS (inductively coupled plasma–mass spectrometry, PerkinElmer, MA, United States) according to the manufacturer’s instructions.