Cryopreserved human hepatocytes

Catalposide (purity, 98%) was obtained from Aobious Inc. (Gloucester, MA, USA). Alamethicin, 3-phosphoadenosine-5-phosphosulfate (PAPS), and uridine 5′-diphosphoglucuronic acid (UDPGA) were from Sigma-Aldrich Co. (St. Louis, MO, USA). 4-hydroxybenzoic acid and 4-hydroxybenzoic acid glucuronide were purchased from Toronto Research Chemicals (North York, ON, Canada). Pooled human intestinal microsomes; pooled human liver S9 fractions; human cDNA-expressed UGTs 1A1/3/4/6/7/8/9/10 and 2B4/7/15/17 supersomes; human cDNA-expressed CESs 1b, 1c, and 2 supersomes; cryopreserved human hepatocytes; and hepatocyte purification kits were obtained from Corning Life Sciences (Woburn, MA, USA). Human cDNA-expressed SULT 1A1*1, 1A1*2, 1A2, 1A3, 1B1, 1C2, 1C4, 1E1, and 2A1 supersomes were purchased from Cypex Ltd. (Dundee, UK). Methanol (HPLC grade) was from Burdick & Jackson Inc. (SK Chemicals, Ulsan, Korea), and all other chemicals were of the highest quality available. Calibration mixtures for Exactive MS [ProteoMass LTQ/FT-hybrid ESI positive mode Cal Mix (MSCAL5) and negative mode Cal Mix (MSCAL6)] were obtained from Supelco (Bellefonte, PA, USA).

Verproside, picroside II, and isovanilloylcatalpol, isolated from Pseudolysimachion rotundum var. subintegrum as described previously [10 (link)], were gifts from KRIBB (Ochang, Korea). Alamethicin, PAPS, William′s E buffer, and UDPGA were obtained from Sigma-Aldrich (St. Louis, MO, USA). Pooled human liver microsomes, pooled human intestinal microsomes, pooled human liver S9 fractions, human cDNA-expressed UGT 1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B15, and 2B17 supersomes, cryopreserved human hepatocytes, and cryopreserved hepatocyte purification kits were obtained from Corning Life Sciences (Woburn, MA, USA). Human cDNA-expressed SULT 1A1*1, 1A1*2, 1A2, 1A3, 1B1, 1C2, 1C4, 1E1, and 2A1 supersomes were obtained from Cypex Ltd. (Dundee, UK). 4-Methylumbelliferone was obtained from Toronto Research Chemicals (North York, ON, Canada). Acetonitrile and methanol (HPLC grade) were obtained from Burdick & Jackson, Inc. (Ulsan, South Korea). The other chemicals were of the highest quality available. ProteoMass LTQ/FT-hybrid ESI Positive mode Cal Mix (MSCAL5) and Negative mode Cal Mix (MSCAL6) for calibration of Q-Exactive MS were obtained from Supelco (Bellefonte, PA, USA).