The iron chelators can be utilized in target cells to improve 5-aminolaevulinic acid (ALA)-based photodynamic therapy (PDT). The purpose of this study is to compare the effect of two kinds of iron chelators, desferrioxamine (DFO) and ethylenediaminetetraacetic acid (EDTA) on the enhancement of ALA-PDT. HaCat cells were cultured in medium containing 2.0 mmol/L of ALA and 0.5 mmol/L of DFO or EDTA. After 3-h incubation in the dark, the concentration of cellular protoporphyrin IX (PpIX) was detected by high performance liquid chromatography (HPLC), and the fluorescence of PpIX was observed at 630 nm emission under confocal laser scanning microscope. For PDT, HaCat cells were irradiated using 632.8 nm laser, and the fractions of apoptotic and necrotic cells were flow cytometrically assayed. Related differences in morphology and ultrastructure of Ha-Cat cells were observed using optical microscope or transmission electron microscope. Compared to incubation with ALA alone, the addition of DFO or EDTA increased the concentration of cellular PpIX and the fluorescent density of PpIX, and also increased cell death ratio after PDT. PDT using ALA plus DFO produced the highest cellular PpIX level, greatest cell death ratio and most severe structural damage to the cells. It was concluded that both DFO and EDTA could enhance ALA-based PpIX production and PDT. Compared to the non-specific iron chelator of EDTA, the specific chelator, DFO, showed more potential for the enhancement.

The iron chelators can be utilized in target cells to improve 5-aminolaevulinic acid (ALA)-based photodynamic therapy (PDT). The purpose of this study is to compare the effect of two kinds of iron chelators,desferrioxamine (DFO) and ethylenediaminetetraacetic acid (EDTA) on the enhancement of ALA-PDT. HaCat cells were cultured in medium containing 2.0 mmol/L of ALA and 0.5 mmol/L of DFO or EDTA. After 3-h incubation in the dark,the concentration of cellular protoporphyrin IX (PpIX) was detected by high performance liquid chromatography (HPLC),and the fluorescence of PpIX was observed at 630 nm emission under confocal laser scanning microscope.For PDT,HaCat cells were irradiated using 632.8 nm laser,and the fractions of apoptotic and necrotic cells were flow cytometricaUy assayed. Related differences in morphology and ultrastructure of HaCat cells were observed using optical microscope or transmission electron microscope. Compared to incubation with ALA alone,the addition of DFO or EDTA increased the concentration of cellular PpIX and the fluorescent density of PpIX,and also increased cell death ratio after PDT. PDT using ALA plus DFO produced the highest cellular PpIX level,greatest cell death ratio and most severe structural damage to the cells. It was concluded that both DFO and EDTA could enhance ALA-based PpIX production and PDT. Compared to the non-specific iron chelator of EDTA,the specific chelator,DFO,showed more potential for the enhancement.

OBJECTIVE：To optimize the microwave processing technology of yellow wine-processed Corydalis yanhusuo . METHODS：The contents of opioid alkaloid ，berberine hydrochloride and tetrahydropalmatine in C. yanhusuo processed with yellow wine were determined by HPLC. The contents of the extracts were determined by hot dipping method. Based on the single factor tests ，using the appearance of yellow wine-processed C. yanhusuo with microwave processing technology ，the contents of extract，opioid alkaloid ，berberine hydrochloride and tetrahydropalmatine as indexes ，with the amount of yellow wine ，wetting time，microwave power and microwaving time as factors ，the processing technology was optimized with orthogonal test combined with comprehensive weighted scoring method ，and then validated and compared with traditional yellow wine-processed C. yanhusuo . RESULTS：The linear ranges of opioid alkaloid ，berberine hydrochloride and tetrahydropalmatine were 0.100-1.500 μg（R2＝0.999 6）， 0.012-0.188 μg（R2＝0.999 5），0.050-0.750 μg（R2＝0.999 8）. RSDs of precision ，stability（12 h）and repeatability tests were all less than 2％ . The recoveries were 99.15％ -100.34％（RSD＝0.54％ ，n＝6），99.52％ -100.78％（RSD＝0.69％ ，n＝6）， 99.26％-99.79％（RSD＝0.28％，n＝6）. The optimum microwave processing technology included that the amount of yellow wine was 4 g（about 20％ of medicinal material amount ），microwave power was 40％，wetting time was 3 hour，processing time was 3 min. The results of three verification tests showed that the contents of extract ，opioid alkaloid ，berberine hydrochloride and tetrahydropalmatine were 15.7％-16.1％，0.061％-0.063％，0.003％-0.004％ and 0.061％-0.063％. The comprehensive scores were 97.916，94.730 and 97.217，and RSD were 0.42％，0.38％，0.46％（n＝3），respectively. Compared with traditional yellow wine processing technology ，there was no significant difference in the contents of opioid alkaloid and other components ，but no scorched spot and crumbs was found in yellow wine-processed C. yanhusuo with microwave processing technology. CONCLUSIONS：Established method for content determination is simple ，accurate，reliable and reproducible ，and can be used for quantitative analysis of active components in yellow wine-processed C. yanhusuo . Optimized microw ave processing technology is stable and feasible ，and can be used for the processing of yellow wine-processed C. yanhusuo .