All reactions involving porphyrin compounds were performed in the dark. Reactions were monitored by thin-layer chromatography (TLC) using aluminum-backed silica gel plates (Macherey-Nagel ALUGRAM® SIL > G/UV254), and ultraviolet light was used to detect TLC spots. Product purifications were performed using Silica Gel 60 (230–400 mesh) for column chromatography and Geduran 60 H Silica Gel (63–200 mesh) for flash column chromatography. Reverse-phase high-performance liquid chromatography (HPLC) was performed on ProStar HPLC (Varian, Agilent Technologies, Santa Clara, CA, USA). Analytical HPLC was done with a Pursuit 5-C18 column (2.5 µm, 4.6×150 mm; Varian) and preparative HPLC with Pursuit 5-C18 column (5 µm, 21.2×150 mm), both using a photodiode array detector (UV–visible detection; Varian) and a fluorescence detector (Varian).
Pursuit 5 c18 column
Manufactured by Agilent Technologies
Sourced in Japan
The Pursuit 5-C18 column is a high-performance liquid chromatography (HPLC) column designed for reversed-phase separation of a wide range of analytes. It features a 5-micron particle size and a C18 stationary phase, providing efficient and reliable separation performance.
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2 protocols using pursuit 5 c18 column
1
Optimized Porphyrin Compound Characterization
2
Characterization of Dexamethasone-loaded Nanoparticles
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A Zetasizer Nano ZS90 (Malvern Instruments, Southborough, MA) was used to determine the particle size, size distribution, and surface charge of DSP-NP. Transmission electron microscope (TEM) images of DSP-NP were obtained using a Hitachi H7600 TEM (Hitachi Co. Ltd., Tokyo, Japan). To measure drug loading, 50 μl of DSP-NP was lyophilized, weighed, and dissolved in 500 μl of acetonitrile. One microliter of 50 mM EDTA was added to solubilize DSP-zinc complex, and the DSP concentration was measured by high-performance liquid chromatography/ultraviolet (HPLC/UV) analysis.
HPLC analysis was performed on a Shimadzu Prominence LC system (Kyoto, Japan) equipped with a Pursuit 5 C18 column (Varian Inc., Lake Forest, CA). Isocratic separation was conducted using the mobile phases consisting of acetonitrile/water [35/65 (v/v)] containing 0.1% trifluoroacetic acid (flow rate = 1 ml/min). Column effluent was monitored by UV detection at 241 nm.
HPLC analysis was performed on a Shimadzu Prominence LC system (Kyoto, Japan) equipped with a Pursuit 5 C18 column (Varian Inc., Lake Forest, CA). Isocratic separation was conducted using the mobile phases consisting of acetonitrile/water [35/65 (v/v)] containing 0.1% trifluoroacetic acid (flow rate = 1 ml/min). Column effluent was monitored by UV detection at 241 nm.
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