Milli q direct water purification system

Methanol (HPLC grade), and formic acid were purchased from Chinese Chemical Reagent Co., Ltd. (Shanghai, China). Ultrapure water (18.2 MΩ·cm⁻¹) was prepared with Milli-Q Direct water purification system (Millipore, USA). Pure creatinine (SRM914a) with a purity of 99.7% was purchased from national institute of standards and technology (NIST), USA. Serum matrix reference materials GBW 09170 and GBW 09171 with certified creatinine values of 8.30 ± 0.10 μg/mL and 35.16 ± 0.39 μg/mL, were developed in our lab (i.e. national institute of metrology, China). Four serum samples, collected from volunteers including two healthy persons and two patients with kidney disease, were provided by the academy of military medical sciences (Beijing, China). All the chemicals were used directly without any further treatment.

Dulbecco’s modified Eagle’s medium (DMEM/High glucose) was purchased from Hyclone (Logan, UT, USA), 0.25% trypsin solution with ethylene diamine tetraacetic acid(EDTA), fetal bovine serum (FBS), and penicillin streptomycin were purchased from Gibco (Grand Island, NY, USA). Detection kits of total antioxidant capacity (T-AOC), thiobarbituric acid reactive substances (TBARS), glutathione peroxidase (GSH-PX), and catalase (CAT) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Assay kits of reaction oxygen species (ROS), cell counting kit-8 (CCK-8), and enhanced bicinchonininc acid (BCA) protein were purchased from Beyotime Biotechnology (Shanghai, China). RNA-easy Isolation Reagent kit was purchased from Vazyme (Nanjing, China). PrimeScript RT (reverse transcription) Master Mix reverse transcription kit, SYBR green PCR kit, and the q225 were purchased from Monad (Suzhou, China). DPPH, quercetin, and HPLC-grade solvents were purchased from Shanghai Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). HPLC-grade water was obtained by filtration using a Milli-Q Direct water purification system from Millipore (Billerica, MA, USA) unless otherwise stated.

Six different types of macroporous resins were bought from H&E Co., Ltd. (Beijing, China). The chemical standards used for HPLC content analysis or HSCCC compound identification were obtained from Chengdu Pure Chem-Standard Co., Ltd. (Chendu, China). HPLC-grade acetonitrile was purchased from Aladdin Industrial Corporation (Shanghai, China). The pure water was prepared by a Milli-Q^® Direct Water Purification System from Millipore Corporation (Bedford, MA, USA). Other analytical-grade solvents used in this study were produced by Lingfeng Chemical Reagent Company (Shanghai, China).

Methanol was obtained from TEDIA (HPLC/Spectro grade, ≥ 99.9%). Acetonitrile was obtained from ACS (HPLC grade, ≥ 99.9%). Water was purified in a MilliQ Direct Water Purification System (Millipore, USA). All standard compounds, including 1,4-Diaminobutane (Put, 99%), spermidine (Spd, ≥99.0%), spermine (Spm, ≥99.0%), 1,4-Diamino(butane-d₈) dihydrochloride (98 atom % D), spermidine–(butane-d₈) trihydrochloride (98 atom % D, 95% CP), spermine–(butane-d₈) tetrahydrochloride (97 atom % D, 95% CP) and heptafluorobutyric acid (HFBA, ≥99.0%) were purchased from Sigma-Aldrich (Hong Kong, China) and used without further purification. Strong Anion Exchange solid phase extraction (SPE) cartridges were obtained from Phenomenex (Strata, 100mg/3mL, USA). Centrifugation was performed using a Refrigerated centrifuge obtained from Eppendorf (5417R, Hong Kong, China).

Fluorescein sodium salt is purchased from Sigma-Aldrich (Milwaukee, WI). The phosphate buffer had a pH of 7.0 and 0.1–100 mM concentrations are used unless otherwise stated. Agarose gel is purchased from Sigma-Aldirch (Milwaukee, WI). Tri-decafluoro-1,1,2,2-tetrahydroctyl-1-trichlorosilane is purchased from United Chemical Technologies (Bristol, PA). 18 MΩcm distilled deionized water is produced with a Millipore Milli-Q^® direct water purification system (Billerica, MA). SU-8 2050 photoresist and SU-8 developer are purchased from MicroChem Corp. (Westborough, MA). The track-etched polycarbonate (TEPC) nanocapillary membrane (NCM) with 10 nm ID pores are purchased from GE water & Process Technologies Inc. (Trevose, PA). The 4-inch silicon wafer were obtained from Addison Engineering (San Jose, CA).

Proton NMR spectra were recorded in CDCl₃ on a 400 MHz Bruker (Billerica, MA, USA) NMR spectrometer using the residual solvent signals (δ_H at 7.28 ppm) as internal standards. High-performance liquid chromatography isolation and purification of compound 1 was conducted on a Shimadzu (Nakagyo-ku, Kyoto, Japan) LC-8A preparative LC coupled to a Shimadzu SPD-M10A VP diode array detector. The UPLC-HRMS/MS analysis was conducted on a Waters (Milford, MA, USA) Xevo G2-XS qTOF with an ESI positive ion mode and data-dependent acquisition mode to obtain MS and MS/MS data. The HPLC-MS/MS analysis was conducted on a Thermo–Finnigan (San Jose, CA, USA) LCQ Advantage ion trap mass spectrometer with an ESI positive ion mode and data-dependent manner to obtain MS and MS/MS data. High-performance liquid chromatography preparative and analytical columns was obtained from Phenomenex (Torrance, CA, USA), and a UPLC analytical column was obtained from Waters. All solvents were HPLC-grade purchased from Thermo-Fisher Scientific (Waltham, MA, USA) and HPLC-grade water obtained by filtration using a Milli-Q Direct water purification system from Millipore (Billerica, MA, USA) unless otherwise stated. Deuterated NMR solvents were purchased from Cambridge Isotope Laboratories (Tewksbury, MA, USA).

Cobaltous nitrate hexahydrate (Co(NO₃)₂·6H₂O), gadolinium nitrate hexahydrate (Gd(NO₃)₃·6H₂O), lanthanum nitrate hexahydrate (La(NO₃)₃·6H₂O), neodymium nitrate hexahydrate (Nd(NO₃)₃·6H₂O), ytterbium nitrate pentahydrate (Yb(NO₃)₃·5H₂O) and samarium nitrate hexahydrate (Sm(NO₃)₃·6H₂O) were obtained from Sigma-Aldrich (USA). Nickel nitrate hexahydrate (Ni(NO₃)₂·6H₂O), strontium nitrate (Sr(NO₃)₂) and zinc nitrate hexahydrate (Zn(NO₃)₂·6H₂O) were obtained from Damao Chemical Reagent Factory (Tianjin, China). Uranium nitrate oxide (UO₂(NO₃)₂) was obtained from Chemical Reagent Purchasing and Supply Station (Shanghai, China). Zirconium chloride (ZrCl₄)) and 2-aminoterephthalic acid were obtained from Heowns Biochemical Technology Co., Ltd (Tianjin, China). Phytic acid sodium salt were purchased fromSolarbio Science & Technology Co., Ltd (Beijing, China). Tetraethoxysilane (TEOS) was obtained from J&K Chemicals Ltd (China). Iron (III) chloride hexahydrate (FeCl₃·6H₂O), Sulfuric acid (H₂SO₄), nitric acid (HNO₃), hydrochloric acid (HCl), ethylene glycol (EG), N, N-dimethylformamide (DMF), methanol (MeOH), ethanol (EtOH), and other reagents were obtained from Tianjin Chemical Reagent No. 6 Factory (China). High-purity deionized water (18.2 MΩ cm) was obtained from a Millipore Milli-Q direct water purification system (USA).