Two chromatographic methods for the quantitative analysis of uridine diphosphate (UDP) sugars involved in hyaluronan pathway of Streptococcus zooepidemicus (SEZ) were developed and compared. The sample preparation protocol using centrifugation and extraction in hot ethanol was employed prior to the analyses. Separation was achieved using an anion exchange Spherisorb SAX column or a Shodex QA-825 column connected with a photodiode array (PDA) detector. To increase the throughput of the chromatography method employing the Spherisorb SAX column, the solid phase extraction (SPE) procedure was introduced. Method validation results displayed that limits of detection (LODs) of UDP-glucose (UDP-Glc), UDP-N-acetylglucosamine (UDP-GlcNAc) and UDP-glucuronic acid (UDP-GlcA) calculated according to QC Expert software were in the low micromolar range and the coefficient of correlation (R(2)) was above 0.997. However, the analytical technique using the Spherisorb SAX column resulted in 80-90% recoveries and low LODs (≤6.19μM), the Shodex QA-825 column showed better long-term stability and reproducible chromatographic properties (RSD≤5.60%). The Shodex QA-825 column was successfully used to monitor UDP-sugar levels during the growth rate of SEZ cells.
- MeSH
- chromatografie iontoměničová MeSH
- intracelulární prostor chemie MeSH
- kyselina hyaluronová metabolismus MeSH
- lineární modely MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- Streptococcus equi chemie metabolismus MeSH
- uridindifosfátové cukry analýza MeSH
- Publikační typ
- časopisecké články MeSH
Physical and chemical structure of paclitaxel (PTX) was studied after its incorporation into polymeric micelles made of hyaluronic acid (HA) (Mw=15 kDa) grafted with C6 or C18:1 acyl chains. PTX was physically incorporated into the micellar core by solvent evaporation technique. Maximum loading capacity for HAC6 and HAC18:1 was determined to be 2 and 14 wt.%, respectively. The loading efficiency was higher for HAC18:1 and reached 70%. Independently of the derivative, loaded HA micelles had spherical size of approximately 60-80 nm and demonstrated slow and sustained release of PTX in vitro. PTX largely changed its form from crystalline to amorphous after its incorporation into the micelle's interior. This transformation increased PTX sensitivity towards stressing conditions, mainly to UV light exposure, during which the structure of amorphous PTX isomerized and formed C3C11 bond within its structure. In vitro cytotoxicity assay revealed that polymeric micelles loaded with PTX isomer had higher cytotoxic effect to normal human dermal fibroblasts (NHDF) and human colon carcinoma cells (HCT-116) than the same micelles loaded with non-isomerized PTX. Further observation indicated that PTX isomer influenced in different ways cell morphology and markers of cell cycle. Taken together, PTX isomer loaded in nanocarrier systems may have improved anticancer activity in vivo than pure PTX.
- MeSH
- antitumorózní látky fytogenní aplikace a dávkování chemie MeSH
- fibroblasty účinky léků MeSH
- HCT116 buňky MeSH
- hydrofobní a hydrofilní interakce MeSH
- isomerie MeSH
- kultivované buňky MeSH
- kyselina hyaluronová aplikace a dávkování chemie MeSH
- lidé MeSH
- micely * MeSH
- nosiče léků aplikace a dávkování chemie MeSH
- paclitaxel aplikace a dávkování chemie MeSH
- polymery aplikace a dávkování chemie MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Hyaluronan (HYA) is a high molecular weight glucosaminoglycan with a great perspective for medical applications. Because HYA is widespread in the body, it is difficult to determine the fate of exogenously administered HYA. METHODS: In this study, HYAof different molecular weights (0.1-1 MDa) was labelled with (99m)Tc, and the distribution profiles were determined after administrating the HYA to rats. RESULTS: After the intravenous administration of (99m)Tc-HYA, a rapid decrease in the radioactivity of blood samples was observed, presumably because of (99m)Tc-HYA uptake by the liver; only minimal signs of liver radioactivity washout were detected. After the oral administration of (99m)Tc-HYA, no significant absorption to the central compartment was found. A preliminary study using (14)C-HYA exhibited a different distribution profile than (99m)Tc-HYA because of the different administered dose and the fate of the degradation products. Even with (14)C-HYA, only traces of radioactivity were absorbed after oral administration. CONCLUSION: This paper provides quantitative information regarding the distribution parameters of radiolabelled HYA in preclinical experiments.
- MeSH
- 99mTc-technecistan sodný MeSH
- aplikace orální MeSH
- gastrointestinální trakt metabolismus MeSH
- gelová chromatografie MeSH
- injekce intravenózní MeSH
- játra metabolismus MeSH
- krysa rodu rattus MeSH
- kyselina hyaluronová krev chemie farmakokinetika MeSH
- ledviny metabolismus MeSH
- molekulová hmotnost MeSH
- orgánová specificita MeSH
- potkani Wistar MeSH
- preklinické hodnocení léčiv MeSH
- radiofarmaka MeSH
- radioizotopy uhlíku MeSH
- stabilita léku MeSH
- tkáňová distribuce MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
