Chloroform

Methanol and n‐hexane (HPLC grade, ≥99.5%) were purchased from Kemiou Chemical Reagent Co., Ltd.. Chloroform (≥99.7%) was purchased from Sinopharm Chemical Reagent. Concentrated sulfuric acid (98%) was purchased from Thermo Fisher Scientific. Lastly, NaCl (≥99.5%) and butylated hydroxytoluene (≥99.8%) were purchased from Tianli Chemical Reagent.

Reagents used in the study are listed in Table I, while the equipment used are listed in Table II. Other chemical agents, including diethylpyrocarbonate (DEPC)-treated water, chloroform, anhydrous ethanol, isopropanol and EDTA, were purchased from Sinopharm Chemical Reagent Co., Ltd., (Shanghai, China).

Monohydroxy-capped poly(ethylene glycol) (PEG₁₁₃-OH) with a mean degree of polymerization, DP, of 113 (M_n = 5000 g/mol, M_w/M_n = 1.02) was purchased from Aldrich and dried at 30 °C under vacuum overnight prior to use. 2-Diethylaminoethyl methacrylate (DEA, Aldrich, 99%) and styrene (St, Aldrich, 99%) were passed through basic alumina column to remove inhibitor and then vacuum distilled from CaH₂ and stored at − 20 °C prior to use. 2, 2-Bipyridine (bpy), copper bromide (CuBr), 2-bromoisobutyryl bromide, 2-aminoethyl methacrylate hydrochloride (AMA^.HCl), d-glucono-δ-lactone, bortezomib (BTZ), and Alizarin Red S (ARS) were all purchased from J&K and directly used. Methanol, toluene, triethylamine (TEA), 1-methyl-2-pyrrolidinone (NMP), and isopropyl alcohol (IPA) were purchased from Sinopharm Group Chemical Reagent Co., Ltd., and then distilled from CaH₂ under reduced pressure. Dichloromethane, chloroform, dimethyl sulfoxide (DMSO), and other organic solvents were purchased from Sinopharm Group Chemical Reagent Co., Ltd., and directly used without further purification. Bromide terminated poly(ethylene glycol) (PEG₁₁₃-Br) macroinitiator and 2-gluconamidoethyl methacrylate (GAMA) monomer were prepared from PEG₁₁₃-OH and 2-bromoisobutyryl bromide, and AMA^.HCl and d-glucono-δ-lactone, respectively, according to literature procedures [35 (link), 36 (link)].

All chemicals and materials used were commercially available and applied without further purification. Iron (III) acetylacetonate (98%), oleic acid (90%), dibenzyl ether were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Platinum (II) acetylacetonate (97%), oleylamine (80–90%), oleyl alcohol, diphenyl ether, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide, N, N’-dicyclohexylcarbodiimide, dopamine hydrochloride, 2-(N-morpholino) ethanesulfonic acid (MES) buffer and rhodamine B isothiocyanate (RITC) were purchased from Aladdin Industrial Inc. (Shanghai, China). Ethanol, hexane, diethyl ether, chloroform, dimethylformamide and sodium carbonate were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). α,ω-dicarboxyl poly(ethylene glycol) (molecular weight = 2000) was acquired from Ponsure Biological Co. (Shanghai, China). Cyclic arginyl–glycyl-aspartic acid (RGD) peptide (cRGDyK) was obtained from GL Biochem (Shanghai) Ltd. Foetal bovine serum (FBS) was purchased from Sijiqing Biologic Co., Ltd. (Hangzhou, China). Dulbecco’s modified eagle medium (DMEM) and trypsin-EDTA solution (0.5% trypsin, 5.3 mM EDTA tetrasodium) were purchased from Jinuo Biomedical Technology Co., Ltd. (Hangzhou, China). Distilled water was prepared by Milli-Q (Millipore, USA).

Live grass carp (2.5–3.0 kg) was purchased from the Huazhong Agricultural University market (Wuhan, China). Arowana multipurpose wheat core wheat flour (Yihai Kerry Arowana Cereals and Oils Food Co., Ltd., Shanghai, China), MT200 modified starch (Tianjin Dingfeng Starch Development Co., Ltd., Tianjin, China), konjac glucomannan (food grade) (Hefei Shengrun Biological Products Co., Ltd., Hefei, China) and rapeseed oil (Shandong Luhua Group Co., Ltd., Yantai, China) were used. Salt and baking powder were purchased from Zhongbai Supermarket of Huazhong Agricultural University (Wuhan, China).
Petroleum ether (30–60 °C), HCl, chloroform, Sudan Red B, acetone, NaBH4, methanol, thiobarbituric acid, disodium EDTA, 1,1,3,3-tetraethoxypropane, formic acid, dichloromethane and hexane were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Carboxymethyl lysine (CML), carboxyethyl lysine (CEL) and methylglyoxalimidazolone (MG-H1) standards were purchased from Toronto Research Chemicals Inc. (Toronto, ON, Canada). Acrylamide standard was obtained from Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA).

An AA500 continuous flow analyzer (SEAL, Germany), anSL252 electronic balance (Shanghai Mingqiao Electronic Instrument Factory, China), and a Milli-Q ultrapure water meter (Merck Millipore, USA) were used. All chemicals used in this work were analyticalgrade, and deionized water was used in all experiments. Hydrochloric acid, sulfuric acid, phosphoric acid, boric acid, chloroform, ethanol, sodium tetraborate, isonicotinic acid, and 4-aminoantipyrine were purchased from Sinopharm Chemical Reagent Co., Ltd. (China). Triton X-100, sodium hydroxide, and potassium chloride were obtained from Tianjin Damao Chemical Reagent Factory (China). Potassium ferricyanide, sodium nitroprusside, sodium salicylate, and N, N-dimethylformamide were supplied by Tianjin Tianli Chemical Reagent Co., Ltd. (China). Potassium dihydrogen phosphate, disodium hydrogen phosphate, pyrazolone, and methylene blue trihydratewere prepared from Tianjin Kemiou Chemical Reagent Co., Ltd. (China). Trisodium citrate dihydrate, polyoxyethylene lauryl ether, and sodium dichloroisocyanurate were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (China). The standard solutions of volatile phenol, cyanide, anionic surfactant, and ammonia nitrogen in water were purchased from the China Academy of Metrology.