Millipore milli q plus system

Raw materials of O. japonicus, L. muscari (named as Shan MaiDong), and L. spicata (named Hubei MaiDong) are listed in Table 1. Their species were identified by Professor Xin Peng, one of the authors of this paper. The samples were stored in the Institute of Chinese Medical Sciences, University of Macau.
2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was purchased from International Laboratory (San Bruno, CA, USA). Potassium persulfate was purchased from Fluka (Selzer, Germany). Dulbecco’s modified eagle medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin (P/S), and phosphate-buffered saline (PBS) were purchased from Gibco-Invitrogen (Paisley, UK). Cell counting kit 8 (CCK8) was purchased from MCE (MedChemExpress LLC, Monmouth Junction, NJ, USA). Griess reagent, fluorescein isothiocyanate-dextran (FITC-Dextran), and lipopolysaccharides (LPS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). An endotoxin detection-specific Limulus test kit was purchased from Bioendo Technology (Xiamen, China). Trifluoroacetic acid (TFA) was purchased from Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA). Nylon membrane filters (0.22/0.45 μm) were purchased from Millipore (Billerica, MA, USA). Deionized water was prepared using a Millipore MilliQ-Plus system (Millipore, Billerica, MA, USA). All the other reagents were of analytical grade.

Chemicals and reagents used in this study were of analytical grade or higher. Ammonia solution, glacial acetic acid, and HPLC grade acetonitrile were purchased from Merck, Darmstadt, Germany. Sodium dihydrogen orthophosphate and sodium hydroxide were obtained from Ajax Finechem Pty Ltd., Auckland, New Zealand. Cyclodextrins, cordycepin, 2′-deoxyguanosine, uridine, and 2′-deoxyadenosine were commercially available from Wako Pure Chemical Industry, Ltd., Osaka Japan. Sodium pyrophosphate decahydrate, adenosine 5′-triphosphate disodium salt (ATP), adenosine 3′-monophosphate (AMP), adenosine 5′-diphosphate disodium (ADP), adenosine, and cucurbit[n]urils were obtained from Sigma-Aldrich Co. St. Louis, MO, USA. Cytidine, 2′-deoxyuridine, inosine, thymidine, and 2′-deoxyinosine were purchased from FUJIFILM Wako Pure Chemical Corporation, Japan. Guanosine was obtained from Alfa Aesar, Thermo Fisher Scientific, Heysham, UK. Acridine orange (AO) and berberine (BE) was purchased from Invitrogen Company, Eugene, Oregon, OR USA. Deionized water was prepared using a Millipore Milli Q-Plus system (Millipore, Bedford, MA, USA). Deuterium oxide (D₂O) was purchased from Cambridge Isotope Laboratories, Inc., Tewksbury, MA, USA.

Dried sea cucumbers H. fuscopunctata Jaeger were purchased from local markets in Guangdong Province and Hainan Province, China. Five oligosaccharide standards from LFG-Na (HS5, pentasaccharide, $M_w$ = 1506 Da; HS8, octasaccharide, $M_w$ = 2462 Da; HS11, hendecasaccharide, $M_w$ = 3417 Da; HS14, tetradecasaccharide, $M_w$ = 4373 Da; HS17, heptadecasaccharide, $M_w$ = 5328 Da) were obtained from Kunming Institute of Botany, Chinese Academy of Sciences. BSA standard was purchased from Sigma (St. Louis, MO, USA). The monosaccharides including L-fucose and N-acetyl-D-galactosamine were purchased from Sigma-Aldrich, and D-glucuronic acid was from J&K Scientific Ltd. (Beijing, China). 1-Phenyl-3-methyl-5-pyrazolone (PMP) was purchased from Xiya Reagnent Co. (Linyi, China). Amberlite FPA98Cl ion-exchange resin was purchased from Rohm and Haas Company (Philadelphia, PA, USA). Deuterium oxide (D₂O, 99.9% Atom D) was obtained from Sigma-Aldrich. The activated partial thromboplastin time (APTT) kits, CaCl₂ and standard human plasma were purchased from MDC Hemostasis (Neufahrn, Germany), BIOPHEN FVIII: C kit was from Hyphen Biomed (Neuville sur Oise, France). Human factor VIII was from Bayer HealthCare LLC. Deionized water was prepared by the Millipore Milli Q-Plus system (Millipore, Billerica, MA, USA). All other reagents were of analytical grade and obtained commercially.

Fisetin, formononetin, daidzein, liquiritigenin, salicylic acid, and ethephon (purity >98%), were purchased from Sigma-Aldrich Company Ltd. (UK). Luteolin was purchased from the National Drug Reference Standards (Beijing, China). D4-SA (purity >99%), which was used as an internal standard, was acquired from Sigma-Aldrich Company Ltd. (UK). Naringenin, Jasmonic acid, and abscisic acid (purity >98%) was purchased from Aladdin Industrial Corporation. (USA). Acetonitrile (HPLC grade) and methanol (HPLC grade) were purchased from Fisher (USA). Other reagents and solvents (analytical grade) were purchased from Beijing Chemical Works (Beijing, China). Purified water using a Millipore Milli Q-Plus system (Millipore, Bedford, MA, USA), was used for making solutions.

Metallurgical aluminum plate (25 × 55 mm, thickness 0.125 mm) (> 99.0%, Goodfellow GmbH) was polished with a 200,00 grit diamond lapidary paste with a diamond powder (size ≤ 0.1 µm) concentration of 50% at the speed of 1000 rpm until a mirror-like surface was observed. The process of surface polishing was carried out in the air. The polished aluminum plate was then cleaned by ultrasound for 10 min in (i) hexane (≥ 99%, Reachem, Slovakia), (ii) 2-propanol (≥ 99.5%, Sigma-Aldrich), (iii) 2 min in Milli-Q water (Milli Q-plus-Millipore system (USA)), then dried under a nitrogen gas stream.