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2 protocols using d saccharic acid 1

1

Comprehensive Characterization of Lipid Excipients

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NOR (purity >98%) was purchased from Chengdu Chroma Biotechnology Co., Ltd. (Chengdu, People's Republic of China). soybean phospholipid (95% phosphatidylcholine) was donated by Shanghai Avet Pharmaceutical Technology Co., Ltd. (Shanghai, People's Republic of China). Labrafac Lipophile WL1349, Labrafil M 1944 CS, Peceol, Capryol 90, Lauroylycol 90, Labrasol, and Transcutol HP (analytical purity) were kindly supplied by Gattefossé (Shanghai, People's Republic of China). Cremophor EL, Cremophor RH 40, and Solutol HS 15 (analytical purity) were donated by BASF (Shanghai, People's Republic of China). Oleic acid and tween 80 (analytical purity) were purchased from Nanjing Chemical Reagent Co., Ltd. (Nanjing, People's Republic of China). Ethyl oleate and MgCl2 (analytical purity) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, People's Republic of China). UDPGA (purity >93%), D-Saccharic acid 1, 4-lactone (purity >98%), and alamethicin (purity >98%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). BCA Protein Assay Kit was purchased from Nanjing Jiancheng Bioengineering Insitute (Nanjing, People's Republic of China). Sinomenine (purity >98%, internal standard) was purchased from Nanjing Jingzhu Bio-technology Co., Ltd. HPLC-grade acetonitrile was purchased from Merck (Darmstadt, German). All other chemicals and solvents were of analytical purity.
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2

Radiolabeled TBBPA Characterization

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[14C]-radiolabeled TBBPA (uniform ring-labeled; Figure 1), previously described by Kuester et al (2007) (link) was generously provided by I. Glenn Sipes at the University of Arizona (Tucson, AZ) for use in the present study. Reversed-phase HPLC fractionation (described below) was used to acquire a radiochemical purity of >98% (specific activity = 90.3 mCi/mmol). [14C]-TBBPA was reconstituted in dimethyl sulfoxide. Chemical purity was determined to be >98% as compared to a TBBPA reference standard (Sigma-Aldrich; St. Louis, MO). Scintillation cocktails were obtained from MP Biomedicals (Ecolume; Santa Ana, CA) or Perkin-Elmer (Ultima-Flo M & PermaFluor E+; Torrance, CA). Cremophore EL®, dimethylsulfoxide, β-glucuronidase (EC 3.2.1.31, Type B-10), ammonium acetate, D-saccharic acid 1, 4-lactone, and aryl sulfatase were purchased from Sigma-Aldrich. All other reagents used in these studies were high performance liquid chromatography (HPLC) or analytical grade.
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