Nexion 350

Normal mice were divided into 2 groups (n = 4 for each group) and IV administrated with CDDP or HSA–CDDP at the same dose and time schedule as the anti-tumor effect examination (3 mg/kg CDDP and equivalent dose of HSA–CDDP as CDDP every other day, seven times). At 72 h after the final IV administration (15 days after first IV administration), mice were euthanized and organs (brain, heart, liver, kidney, spleen, and lung) and blood were acquired and weighed. For tumor CDDP distribution, U87MG or U87MG-shSPARC cells (2 × 10⁶ cells/site) were injected subcutaneously into the right lower flanks. When tumor volume approached 50 mm³, mice were randomly divided into 2 groups (n = 5 for each group) and IV administrated CDDP or HSA–CDDP (every other day, seven times). At 72 h after the last IV administration (15 days after first administration), mice were euthanized, and the tumor was acquired. All samples (organs, blood, and tumors) were lyophilized and we analyzed the amount of CDDP using the ICP-MS (NexION 350; Perkin-Elmer, Waltham, MA, USA) installed at the National Center for Inter-university Research Facilities (NCIRF) at Seoul National University.

Induced Coupled Plasma Mass Spectrometry (ICP-MS) NexION 350 by Perkin Elmer Inc. (Waltham, MA, USA) is used to assess the concentration of Gd-DTPA loaded within the NPs. A suspension of CS-HA nanoparticles in deionized water at a concentration of 250,000 particles/mL without dissolution is injected. All data are collected and processed using the Syngistix NanoApplication (Module PerkinElmer Inc., Waltham, MA, USA). Gd is measured at m/z 157 using a 100 μs dwell time with no settling time. Successively, results are compared to the known initially added amount of Gd-DPTA to obtain an estimate of the encapsulation efficiency.

Of sediment, 0.5 g was weighed and 10 mL of 7 M HNO₃ was added and the reaction mixture was autoclaved at 121 °C and 200 kPa for 30 min (SIS 1993 ). After cooling, the samples were centrifuged for 20 min at 3000 g and 20 °C. After centrifugation, the supernatant was decanted carefully and stored in a 50-mL acid washed tube; the remaining sediment was discarded.
The mineral fractions obtained during each extraction step (fractions 1 through 4) and pseudo-total metal concentrations were analyzed on an ICP-MS (Perkin Elmer NexION 350). Prior to analysis, an internal standard was added, and the samples were diluted with deionized water. To evaluate the reliability of the extraction procedure, the reference standard CRM-601 was extracted, and analyzed for six certified trace metal concentrations in similar manner as the sediment samples.

The lichen samples were air-dried, cleaned from any extraneous material under a binocular microscope, and pulverized using mortar and pestle and liquid nitrogen. Approximately 250 mg of lichen powder was dissolved in 3 mL of 70% HNO₃, 0.2 mL of 60% HF, and 0.5 mL of 30% H₂O₂ in a microwave digestion system (Milestone Ethos 900) at 280 °C and 55 bar (Loppi, 2014 (link)). The concentrations of selected PTEs, namely Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sb, and Zn, were determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS, Perkin Elmer NexION 350). The quality of analytical procedures was verified using procedural blanks and the certified material IAEA-336 “Lichen” (International Atomic Energy Agency, 1999 ), which indicated recoveries in the range of 93–105%. The results are expressed on a dry weight basis (mg/kg dw). For each species, five replicates were measured.

Cells were seeded in a 6-well plate and incubated. When the cells were in the logarithmic growth phase, aspirate the medium, 2 ml of medium containing CoNPs (400 μM), and ALA (200 μM) were added to each well. After 24 h of incubation, 1 ml of the upper-medium was drawn before collecting the cells to measure the concentration of cobalt in the extracellular solution. Then, the medium was removed, and cells were washed with PBS 3 times, detached, and collected with centrifugation at 1000 rpm for 5 min. 2 mL of PBS was used to re-suspend the cells. After 5 min, 10 uL of supernatant was extracted to check with a microscope to confirm there are no CoNPs in the solution. Next, 1 × 106 cells were again centrifuged at 1000 rpm for 5 min. After removing the supernatant, 500 μL of lysis buffer was added, followed by centrifugation at 12,000 rpm for 2 h. Finally, the supernatant was used to measure the cobalt concentration with an inductively coupled plasma mass spectrometer (ICP-MS, PerkinElmer NexION 350).

An automated biochemical analyzer (Hitachi 7600, Tokyo, Japan) was used to evaluate the concentrations of serum fasting glucose, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), and triglyceride (TG). And glycated hemoglobin (HbA1c) was measured using HPLC (Waters E2695, Milford, MA, USA).
Inductively coupled plasma mass spectrometry (ICP-MS, PerkinElmer, NexION 350, Waltham, MA, USA) was used to detect the amounts of magnesium and calcium. The plasma element was measured by 0.5% (v/v) high-purity nitric acid dilution (1:20). The precision and accuracy of the analysis were checked at 10-sample intervals using the quality control samples (Seronorm, Level-2, Billingstad, Norway).
The coefficient of variation for Mg was 2.33% for between batches and 1.19% for within batches. The coefficient of variation for Ca was 1.23% for between batches and 2.62% for within batches. The recovery of Mg and Ca were 100.10% and 97.63%, respectively.