ABSTRACT
Objective: To establish a quick method of ultra-performance liquid chromatography-quadrupole time-of-fight mass spectrometry (UPLC-Triple-TOF/MS) for the analysis of components of crude and sweated Dipsaci Radix. Methods: The separation was performed on the chromatographic column of Agilent Eclipse XDB-C18 (250 mm × 4.6 mm, 5.0 μm), and the mobile phase was 0.1% formic acid solution-methanol, with a gradient elution at a flow rate of 0.8 mL/min, the detection wavelength was 215 nm, the column temperature was 25 ℃. UPLC-Triple-TOF 5600+ time of flight liquid and mass spectrometer was used for mass spectrometry. Electrospray ion source negative ion mode was adopted, and the scanning range was m/z 100-1 500. The components of crude and sweated Dipsaci Radix were quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results: Fifty-two common components were identified or tentatively characterized based on the retention time and MS spectra. They were triterpenoid saponins, iridoids, phenolic acids etc. The crack rules of primary components were also analyzed. And comparing the components of crude and sweated Dipsaci Radix, it showed that the content of 20 components such as loganin acid, chlorogenic acid, loganin, isochlorogenic acid A, and asperosaponin VI was decreased after sweating, and caffeic acid, isochlorogenic acid, isochlorogenic acid C, and triplostoside A was increased. Conclusion: The types of components of crude and sweated Dipsaci Radix are identical, but there are differences in the content of the components. The content of the components of crude are higher than the sweated Dipsaci Radix. UPLC-Triple-TOF-MS technology was used to analyze the influence of “sweating” on the chemical composition of the Dipsaci Radix, so as to provide a theoretical basis for the study of the chemical constituents of sweated Dipsaci Radix and further research on the origin processing of Dipsaci Radix.
ABSTRACT
<p><b>BACKGROUND</b>We previously reported that iodine-131((131)I)-labeled anti-pro-gastrin-releasing peptide (ProGRP(31-98)) monoclonal antibody D-D3 could selectively accumulate in the tumor sites of nude mice bearing small cell lung cancer (SCLC) xenografts. However, (131)I-D-D3 was cleared slowly from the body, and the best radioimmunoimaging time for SCLC was 72 - 96 hours after injection. The aims of this study were to radiolabel anti-ProGRP(31-98) D-D3 monoclonal antibody with technetium-99m ((99m)Tc) and to investigate the biodistribution of this antibody in healthy ICR mice.</p><p><b>METHODS</b>D-D3 was labeled with (99m)Tc via the 2-mercaptoethanol reduction method. (99m)Tc-D-D3 was purified by the gel column separation method. The labeling efficiency and radiochemical purity were measured by thin-layer chromatography. The immunological activity of (99m)Tc-D-D3 was determined with cell conjugation assays. (99m)Tc-D-D3 was injected into healthy ICR mice via a tail vein, and all the healthy ICR mice were sacrificed by cervical dislocation at a designated time. Then, the blood and major organs were removed and weighed, and counted in a gamma scintillation counter to determine the percentage of the injected dose per gram (%ID/g).</p><p><b>RESULTS</b>The labeling rate and the radiochemical purity of (99m)Tc-D-D3 were (73.87 ± 2.89)% and (94.13 ± 4.49)%, respectively. The immunobinding rates of (99m)Tc-D-D3 to the human small cell lung cancer NCI-H446 cell line and lung adenocarcinoma A549 cell line were (81.2 ± 2.37)% and (24.3 ± 1.46)%, respectively. The distribution data of normal ICR mice demonstrated that (99m)Tc-D-D3 was mainly distributed in the liver, kidney and lung, and less in the brain tissue and muscle.</p><p><b>CONCLUSIONS</b>(99m)Tc-D-D3 antibody not only had high radiochemical purity, but also had good stability both in vitro and in vivo, and maintained good immunological activity. (99m)Tc-D-D3 was metabolized mainly in the kidney and liver, and the blood radioactivity decreased rapidly. Thus, (99m)Tc-D-D3 is conducive to the radioimmunoimaging of SCLC.</p>