Quantitative high-performance liquid chromatography-based detection method for calphostin C, a naturally occurring perylenequinone with potent antileukemic activity
| Autor: | Hao Chen, Fatih M. Uckun, De-Min Zhu, Chun-Lin Chen |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Detection limit
Antibiotics Antineoplastic Chromatography Chemistry Elution Spectrum Analysis Reproducibility of Results General Chemistry Naphthalenes Precursor Cell Lymphoblastic Leukemia-Lymphoma Sensitivity and Specificity High-performance liquid chromatography Blood proteins Mice chemistry.chemical_compound Calphostin C In vivo Trifluoroacetic acid Animals Female Enzyme Inhibitors Quantitative analysis (chemistry) Chromatography High Pressure Liquid Protein Kinase C |
| Zdroj: | Journal of Chromatography B: Biomedical Sciences and Applications. 724:157-162 |
| ISSN: | 0378-4347 |
| Popis: | Calphostin C is a potent inhibitor of protein kinase C and can induce Ca2+-dependent apoptosis in human ALL cells. Further development of calphostin C will require detailed pharmacodynamic studies in preclinical animal models. Therefore, we established a sensitive and accurate high-performance liquid chromatography (HPLC)-based quantitative detection method for the measurement of calphostin C levels in plasma. Extraction of calphostin C from plasma was performed by precipitation of plasma protein using acetonitrile and an aliquot of extracted supernatant was injected onto a Hewlett-Packard HPLC system constituting a 250x4 mm LiChrospher 100, RP-18 (5 microm) in conjunction with a 4x4 mm LiChrospher 100, RP-18 guard column (5 microm). The eluted compounds were detected by diode array detection set at a wavelength of 479 nm. Acetonitrile-water containing 0.1% trifluoroacetic acid and 0.1% triethylamine (70:30, v/v) was used as the mobile phase. The average extraction recovery from plasma was 97.3%. Good linearity (r0.999) was observed throughout the concentration range of 0.05-40 microM for calphostin C in 50 microl of plasma. Intra- and inter-assay variabilities were less than 6% in plasma. The lowest detection limit of calphostin C in 50 microl plasma was 0.02 microM at a signal-to-noise ratio of approximately 3. The availability of this assay will now permit detailed pharmacodynamic and pharmacokinetic studies of calphostin C in vivo. |
| Databáze: | OpenAIRE |
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