Agilent 1200 infinity series

The analyses of catechin, 2,3,4,5ʹ-tetrahydroxystilbene-2-O-α-glucoside (TSG), rhein, emodin, and chrysophenol were performed on an Agilent 1200 Infinity series (Agilent Technologies, Santa Clara, CA, USA). Samples were separated on an XTerra RP18 analytical column (4.6 × 250 mm, 5 μm; Waters Corp., Milford, MA, USA). The mobile phase contained 0.5% glacial acetic acid in water (solution A) and 0.5% glacial acetic acid in acetonitrile (solution B). The injection volume was 20 μL, and the flow rate was 1.0 mL/min at 30 °C. The detection wavelength was 280 nm.

The plasma homocysteine was determined by HPLC-MS/MS (Agilent 1200 Infinity Series, Agilent Technologies, Santa Clara, CA, USA; Triple Quad™ 4500, SCIEX, Framingham, MA, USA). The coefficient of variation was of 1.5–1.6%.

The deionized water from EASY pure LF, Barnstead, was used to prepare all the solutions. The characterization of ZHN-SDS-BP was carried out with a scanning electron microscope (SEM) and a transmission electron microscope (TEM) using a field emission scanning electron microscopy (FESEM) model Hitachi SU 8020 UHR (Tokyo, Japan). The pH measurements were conducted with a pH meter, an Orion 720A (MI, USA), equipped with a glass electrode. Cyclic voltammetry (CV) and square-wave voltammetry (SWV) were performed with the Potentiostat Gamry Series-G750 (Warminster, PA, USA), and the electrochemical data were obtained using three-electrode systems consisting of an Ag/AgCl electrode model MF-2052 from Bioanalytical System (West Lafayette, IN, USA) with a fiber junction as the reference electrode, a platinum wire as a counter/auxiliary electrode, and a ZHN-SDS-BP-modified multi-walled carbon nanotube (MWCNT) paste electrode as a working electrode. The electrochemical impedance spectroscopy (EIS) measurements were carried out on a Potentiostat/Galvanostat Gamry model Ref 3000 (PA, USA). Convincingly, the real sample validation was conducted by using high-performance liquid chromatography (HPLC, column, C-18; detector, UV-Vis (DAD) 280 nm) with the Agilent 1200 Infinity Series (Waldbronn, Germany).

Plasma metabolic fingerprinting in the RH study was performed with the Agilent 1200 Series LC system (Agilent Technologies, Waldbronn, Germany) coupled with the Agilent 6224 Series TOF LC/MS system (Agilent Technologies, Waldbronn, Germany). In the PH study, plasma metabolic fingerprinting was conducted with the Agilent 1200 Infinity series (Agilent Technologies, Waldbronn, Germany) coupled with the Agilent Technologies QTOF (6520) mass spectrometry detector. The chromatographic and mass spectrometer parameters of the optimized LC/MS methods were described in detail in the Supplementary Material section. Quality control samples (QCs) were prepared as a pool of equal volume of each plasma samples included in each study. The QCs were analyzed in order to monitor system's and method's stability during the whole sequence run. Detailed clinical information about studied groups in both non-targeted metabolomics studies were described in Tables S1, S2 in the Supplementary Material section.

The plasma methylmalonic acid was determined by HPLC-MS/MS (Agilent 1200 Infinity Series, Agilent Technologies, Santa Clara, CA, USA; Triple Quad™ 5500, SCIEX, Framingham, MA, USA). The coefficient of variation was of 2.6–5.0%.

An SEC-HPLC method was developed to analyze
the stability and homogeneity of ScpA variants using an Agilent 1200
Infinity Series (Agilent Technologies, USA) and comprised a quaternary
pump (G1311B 1260), an ALS autosampler (G1329B 1260), a thermostated
column compartment (G1316A 1260), and an MWD VL diode-array detector
(G1365D 1260). The chromatographic separations were carried out using
an AdvanceBio SEC 300A 2.7 μm 7.8 × 300 mm column. The
column was washed with water and equilibrated with the mobile phase
(150 mM sodium phosphate, pH 7.0) for 2 h with a flow rate of 1 mL/min.
The ScpA samples (5 μL of a 1 mg/mL solution) were injected,
and the eluate was analyzed for 15 min, using both 280 and 220 nm
detection wavelengths. The peak shape and retention times were analyzed
and used for the interpretation of purity. All the measurements were
performed in triplicates and the average spectra used for analysis.