7800 icp ms

The metalloproteomes for each condition were analyzed via SEC-ICP-MS using an Agilent Infinity II LC coupled to an Agilent 7800 ICP-MS. The methods were inspired by work done by the Roberts lab [18 (link),19 (link)]. A total of 160 µg of soluble protein (entire proteome) was injected onto an Agilent Bio-SEC 3 (3 µm, 300 Å, 4.6 × 300 mm) column and proteins were separated for 25 min using 200 mM ammonium acetate pH 8 as the mobile phase, with a flow rate of 0.4 mL/min. The five metal ion signals were monitored, and these included: ²⁴Mg, ⁵⁶Fe, ⁶³Cu, ⁶⁶Zn, and ⁷⁵As, with an integration time/mass of 1.5 s per analyte. The monitoring of ion signals was performed using the Agilent MassHunter 4.6 (version C.01.06). Metal signals were manually integrated in MassHunter, and all other data workup was performed in Microsoft Excel. Statistical differences were determined using a Student’s t test at a 95% confidence interval.

Proximate analyses were performed following the standard methods (official method, OM) of the Association of Official Analytical Chemists (AOAC, 2005) [78 ]. Approximately 1.0 g of the powdered plant samples (AAL, AAS, AVL, AVS, AVR, ATL, ATS, and ATR) were used to estimate total moisture (OM 930.04), ash (OM 930.05, using muffle furnace Model: KAA/956/C, Sri Krishnaa Enterprises, Secunderabad, Telangana, India), crude fat (OM 2003.05, extraction unit Model: E–816 HE, BÜCHI Labortechnik AG, Flawil, Switzerland), crude protein (OM 977.02 using Kjeldahl factor of 6.25, distillation unit Model: VAPODEST 200, C. Gerhardt GmbH & Co. KG, Königswinter, Nordrhein-Westfalen, Germany), and carbohydrate contents. The carbohydrate content was calculated as the difference from the other components analyzed. Heavy metals and minerals compositions were also determined to compare the nutritional status of the samples (OM 2015.01 (2018) using inductively coupled plasma mass spectrometry Model: 7800 ICP-MS, Agilent Technologies, Santa Clara, CA, USA) [79 (link)].

To simulate the effect of 7-day degradation of Mg²⁺ production on MC3T3-E1 cells, we prepared α-MEM with varying Mg²⁺ concentrations. First, inductively coupled plasma mass spectrometry (7800 ICP-MS, Agilent, United States) was performed to detect the Mg²⁺ concentration of pure Mg and Mg-Ti composite extracts. Then, the sterilized Mg chloride solution was applied to elevate the α-MEM Mg²⁺ concentration, according to the result of the Mg²⁺ concentration test.

The mineral content was measured by digesting the samples in a microwave oven (MARS 6 iWave, CEM Corporation, Mattews, NC, United States of Americawith sulfuric acid, hydrogen peroxide, and hydrochloric acid [30 (link)]. This solution was atomized in argon plasma, and due to the high temperature, the sample dried further and was ashed, atomized, and ionized. The minerals were detected using an ICP-MS (Agilent 7800 ICP MS, Matthews, NC, USA), and the signal intensity of the mineral was divided by the charge of the minerals [31 ]. The mineral content was quantified by means of a calibration line based on the ratio between the signal of the element and the signal of the associated standard.

In vitro degradation profiles and microstructure evolutions of Fe/Ce-MSN NPs were assessed by two typical approaches. One was detecting the degradation content of Fe, Ce and Si by inductively coupled plasma mass spectrometry (ICP-MS, 7800 ICP-MS, Agilent), and the other was directly observing the time-dependent structural evolution of Fe/Ce-MSNs by SEM during the degradation evaluation.
Typically, 1 mg Fe/Ce-MSNs was added into 1 ml FBS (PBS as control) solution with different pHs (pH: 7.4 and 6.0) for 24 h. The testing solution was put into a water bath at 37°C under magnetic stirring slowly (250 rpm). ICP-MS test and SEM images of samples were taken out at different time points to monitor the degradation.

The pH (pH meter, Mettler Toledo, Mumbai, India), total soluble solids (TSS), and titrable acidity of the prepared beer samples were determined²⁶ . HPLC was used to determine the amount of ethanol and sugars. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) was used to examine the minerals (7800 ICP MS, Agilent Technologies, USA). Total phenol content was determined using Kumari et al.^{27 (link)} methodology at 650 nm with a spectrophotometer (UV3000 +, Lab India, Delhi, India). The total monomeric anthocyanins (TMA) content was determined using two different wavelengths of 520 and 700 nm^{28 (link)}. Color was determined using EBC (European Brewing Convention) units at 430 nm^{29 (link)}.

Samples were collected to determine the concentrations of DPrP, its metabolites, and Cd(II) during incubation. Extraction and detection of DPrP were carried out according to the method described previously [23 (link)]. The metabolites produced during the degradation process were identified by HPLC-MS [24 (link)]. The amino acids and fatty acids in the supernatant were detected by LC-MS [25 (link)] and GC-MS [26 (link)], respectively. The adsorption of Cd(II) on the surface and inside of bacteria was determined by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy dispersive spectroscopy (EDS) [27 (link)]. The Cd(II) concentration in the supernatant was investigated by ICP-MS (Agilent Technologies 7800 ICP-MS) analysis [28 (link)]. Quantitative PCR (qPCR) was used to quantify the ratio of DNA levels of the specific estG/xcpR genes to determine the ratios of ZM05 to ZM03 in the coculture system under Cd(II) stress [29 (link),30 (link)]. The primers used in qPCR are listed in Supplementary Table S1. For the transcriptional analysis, CK (control), CD (monoculture under 0.8 mM Cd²⁺), and CO (coculture under 0.8 mM Cd²⁺) samples were harvested for RNA extraction, and sample information is provided in Table S2.