Functional group: ADME NEW from Shiseido CAPCELL PAK INERT ADME CAPCELL PAK INERT ADME S3 2 4 6 8 10 12 [min] EXCELLENT Peak Shape ! Diphacinone M.W. : 340.37 Conventional C18 column S3 【HPLC Conditions】 Column size : 2.0 mm i.d. x 50 mm Mobile phase : 10 mmol/L HCOONH4 / CH3CN = 70 / 30 Flow rate : 0.2 mL/min Temperature : 40 ℃ Detection : UV 285 nm Inj. vol. : 1 mL Sample dissolved in : 1 mg/mL in CH3CN, diluted with 20% CH3CN at 10 mg/mL. 2 4 6 8 10 12 [min] CAPCELL PAK INERT ADME S3 AMP Large Retention Times for Polar Compounds! k = 1.71 [min] 1 2 3 4 5 6 ADP k = 0.56 ATP k = 0.20 ATP M.W. : 507.18 【HPLC Conditions】 Column size : 2.0 mm i.d. x 150 mm Mobile phase : 10 mmol/L HCOONH4 / CH3CN = 99 / 1 Flow rate : 0.2 mL/min Temperature : 40 ℃ Detection : UV 260 nm Inj. vol. : 1 mL Sample dissolved in : H2O (10 mg/mL each.) k = 1.05 k = 0.31 Competitor’s metal-free ODS column 3 mm k = 0.12 Realized by: Analyte Strong interaction Inert column Functional group: ADME Inner surface : PEEK Filter : PEEK

Characteristics Severe peak tailings or no peaks !? Adsorbed to the column ? Metal coordination suspected ? Hard to retain hydrophilic compounds ? Cephem antibiotics　～Ceftriaxon～ Ceftriaxon M.W. : 554.59 Ceftriaxon CAPCELL PAK C18 MGIII-H S3 [min] 1.0 2.0 3.0 4.0 5.0 CAPCELL PAK INERT ADME S3 CAPCELL PAK ADME S3 １ 3 4 5 6 7 5,6 CAPCELL PAK INERT ADME S3 0.0 1.0 2.0 3.0 4.0 [min] S/N = 20.5 0.0 1.0 2.0 3.0 4.0 [min] S/N = 10.1 CAPCELL PAK ADME S3 MRM　555->396 Nucleotide 【HPLC Conditions】 Column Size : 2.0 or 2.1 mm i.d. × 50 mm　 Mobile phase : A) 0.1 vol% HCOOH, B) 0.1 vol% HCOOH, CH3CN PDA ; B 15% -> 95% (3 min) MS ; B 15% -> 95% (2 min) Flow rate : 0.5 mL/min Temperature : 40˚C Detection : PDA 260 nm, MS ESI positive Inj.vol. : 1 mL Sample : PDA 20 mg/mL each. MS 200 ng/mL 1 2 3 4 5 6 Conventional C18 column S3 1. Cephapirin 2. Ceftriaxone 3. Cefazolin 4. Cephaperazone 5. Cephalothin 6. Sefaca pen Pivoxil 7. Cefditoren pivoxil 1 2 3 4 5 6 [min] 346->79　(AMP) 426->79　(ADP) 506->159 (ATP) N.D. ATP N.D. Characteristics 【HPLC Conditions】 Column size : 2.0 mm i.d. x 150 mm Mobile phase : 10 mmol/L HCOONH4 / CH3OH = 98 / 2 Flow rate : 0.2 mL/min Temperature : 40 ℃ Detection : MS ESI negative Inj. vol. : 2 mL Sample dissolved in : H2O (150 ng/mL each.) CAPCELL PAK INERT ADME S3 *　All date acquired by NANOSPACE 2 HPLC system (Inert specification). P/N I.D (mm) Length (mm) 79001 2 50 79002 100 79003 150