TY - JOUR
T1 - Cytochalasin B
T2 - Preparation, analysis in tissue extracts, and pharmacokinetics after intraperitoneal bolus administration in mice
AU - Lipski, Karen M.
AU - McQuiggan, James D.
AU - Loucy, Karen J.
AU - Fondy, Thomas P.
N1 - Funding Information:
Technical assistance during all aspects of this work was provided by Lesa A. Paulsen. Extractability of cytochalasin A was determined by Mehran Massoumi-Sourey. We are grateful to Dr. James R. Florini for use of HPLC instrumentation and to Dr. Grant Krafft for assistance with NMR spectra and with conditions for HPLC determinations. NMR instrumentation was supported by NIH Grant RPO 13 I7 to Syracuse University, Department of Chemistry.
Funding Information:
Kaplan. ’ Supported by the Stella Hardeman Grant for Cancer Research to the American Cancer Society. ’ To whom correspondence should be addressed.
PY - 1987/3
Y1 - 1987/3
N2 - Cytochalasin B (CB) was prepared by methanol extraction of dehydrated mold (Drechslera dematioidea) matte, reverse-phase C18 silica gel batch adsorption, selective elution with 1:1 (v/v) hexane:tetrahydrofuran (THF), crystallization, preparative TLC, and recrystallization. Unit gravity silica gel normal phase chromatography afforded additional CB. Yield per liter of medium was 300 mg of CB >95% pure by NMR, HPLC (60:40 hexane: THF, Lichrosorb Si60 silica gel, 230 nm), and TLC. CB added exogenously to mouse organs at 1 and 5 μg/organ was recovered 70 to 100% by methanol extraction, adsorption to C18 silica gel Sep-Pak cartridges, elution with ethyl acetate, and analysis by TLC and/or HPLC. Limiting sensitivity (micrograms/extract) was 0.5 TLC; 1.0 HPLC. Quantitative extraction was confirmed with 3H-labeled CB. CB ip in mice at 50 mg/kg (LD10) distributed rapidly into liver, renal fat, kidney, intestines, mesentery, pancreas, spleen, and blood cells and was cleared from all but liver within 24 h. CB was below detectable levels in thymus, lymph nodes, heart, brain, bone marrow, and lungs. Cytochalasin A is fixed to tissues and not extractable. This work affords a source of CB in quantities permitting in vivo study, provides methods for extraction and analysis, and reveals the pharmacokinetics of ip bolus CB.
AB - Cytochalasin B (CB) was prepared by methanol extraction of dehydrated mold (Drechslera dematioidea) matte, reverse-phase C18 silica gel batch adsorption, selective elution with 1:1 (v/v) hexane:tetrahydrofuran (THF), crystallization, preparative TLC, and recrystallization. Unit gravity silica gel normal phase chromatography afforded additional CB. Yield per liter of medium was 300 mg of CB >95% pure by NMR, HPLC (60:40 hexane: THF, Lichrosorb Si60 silica gel, 230 nm), and TLC. CB added exogenously to mouse organs at 1 and 5 μg/organ was recovered 70 to 100% by methanol extraction, adsorption to C18 silica gel Sep-Pak cartridges, elution with ethyl acetate, and analysis by TLC and/or HPLC. Limiting sensitivity (micrograms/extract) was 0.5 TLC; 1.0 HPLC. Quantitative extraction was confirmed with 3H-labeled CB. CB ip in mice at 50 mg/kg (LD10) distributed rapidly into liver, renal fat, kidney, intestines, mesentery, pancreas, spleen, and blood cells and was cleared from all but liver within 24 h. CB was below detectable levels in thymus, lymph nodes, heart, brain, bone marrow, and lungs. Cytochalasin A is fixed to tissues and not extractable. This work affords a source of CB in quantities permitting in vivo study, provides methods for extraction and analysis, and reveals the pharmacokinetics of ip bolus CB.
KW - HPLC drug metabolites
KW - cancer chemotherapy
KW - drug analysis
KW - metabolites drugs
KW - thin-layer chromatography
KW - tissue homogenization
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U2 - 10.1016/0003-2697(87)90459-3
DO - 10.1016/0003-2697(87)90459-3
M3 - Article
C2 - 3578799
AN - SCOPUS:0023148859
SN - 0003-2697
VL - 161
SP - 332
EP - 340
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -