OBJECTIVETo develop a qualitative screening method by high performance liquid chromatograph-photo-diode array (HPLC-PDA) and a confirmation method by ultra performance liquid chromatograph-quadrupole-time of flight mass spectrometry (UPLC-Qtof-MSMS) for Copper Chlorophyll in olive oils.

METHODSCommercial liposoluble copper chlorophyll diluted by petroleum ether was purified and concentrated by sep-pak silica solid-phase extraction (SPE) cartridges, the injection was separated by symmetry shield RP18 column, and detected by the PDA at the wavelength 430 nm. The injection was separated by ACQUITY UPLC BEH (18)C and determined by UPLC-Qtof-MSMS in negative atmospheric pressure chemical ionization mode, then confirmed by the exact mass of parent ion and secondary fragment ion provided by the mass spectrum. Samples from supermarket were screened for Cu-pyropheophytin a by established method.

RESULTSCu-pyropheophytin a could be used for discrimination as indicative compound whether the olive oils were mixed with copper chlorophyll. 25 samples including olive oil, soybean oil and spirulina from supermarket were screened, Cu-pyropheophytin a was found in one type of olive oil.

CONCLUSIONThis method could be used for screening and qualitative analysis of Copper Chlorophyll in olive oils for its accuracy , reliability and excellent specificity.

Chlorophyllides ; Chromatography, High Pressure Liquid ; Food Contamination ; Mass Spectrometry ; Olive Oil ; Plant Oils ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo evaluate the efficacy and safety of sodium copper chlorophyllin (trademarked as "Yebaike Tablet which is abbreviated as YBK in treating leukopenia.

METHODSOne hundred and five patients with leukopenia caused by various factors were randomized into 3 groups. The 60 patients in the YBK group took orally YBK Tablets at the dose of 40 mg, three times per day, the 30 patients in the leucogen group were treated with Leucogen Tablets at the dose of 20 mg, three times per day, and the 15 patients in the placebo group were administered with vitamin C tablets 100 mg, three times per day. All the subjects were treated for 1 month. The change of peripheral leucocytes count after treatment and adverse drug reaction that occurred in patients were studied.

RESULTSIn the 60 patients treated with YBK, the treatment proved to be markedly effective in 34 cases, effective in 17 and ineffective in 9, the total effective rate being 85%, which was significantly higher than that in the placebo group (26.7%, P < 0.01) and similar to that in the leucogen group (83.3%, P > 0.05). No adverse reaction was found in the treatment course.

CONCLUSIONYBK can be used in the treatment of leukopenia caused by various factors, satisfactory in efficacy and safe in use.

Adolescent ; Adult ; Ascorbic Acid ; administration & dosage ; Chlorophyllides ; administration & dosage ; adverse effects ; Female ; Humans ; Leukocyte Count ; Leukopenia ; drug therapy ; Male ; Middle Aged ; Tablets ; Thiazoles ; administration & dosage ; Thiazolidines ; Treatment Outcome

The blood-brain barrier (BBB) is a huge obstacle in therapy of brain diseases, for it hinders the delivery of water-soluble molecules and those with molecular weight above 500 from the circulation system to the brain. Polysorbate 80 (Tween 80, T-80)-coated polylactid acid(PLA) nanoparticles represent a tool to transport such drugs across the BBB. Transcytosis is put forward as one mechanism of drug-loaded nanoparticles across the blood-brain barrier (BBB). However little is known about it. Electron microscopy is an important method in the investigation on nanoparticles injected into the experimental mice. In this study it was found by fluorescence microscope that fluorescence existed along the capillary dissepiment. Some nanoparticles distributed in the brain capillary endothelial cells and brain tissue outside the microvaculum, which was observed by transmission electron microscopy. These particles were proved to be the Copper chlorophyll loaded nanoparticles which containing Cu detected by AEM. The in vivo experiments demonstrated directly that the PLA nanoparticles can pass the BBB indeed and transcytosis by microvascular endothelial cells may be the mechanism. The results provided an efficient way of drug-delivery targeting the brain. Copper chlorophyll could be used as a new symbol of nanoparticles in in vivo experiment.
Animals ; Blood-Brain Barrier ; Chlorophyllides ; Lactic Acid ; pharmacokinetics ; Mice ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Nanoparticles ; Polyesters ; Polymers ; pharmacokinetics ; Spectrometry, X-Ray Emission

OBJECTIVETo study the inhibitory effect of chlorophyllin (CHL) on trans-benzo(a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide (BPDE) induced malignant transformation in human bronchial epithelial cell line (16HBE).

METHODS10, 50 or 100 micro mol/L CHL were added into the media during the cells transformation induced by BPDE, and the malignant degree of transformed cells were identified by the ConA agglutination test and the assay for anchorage-independent growth and tumorigenicity.

RESULTSAfter the cells were cultured for 25 times, the time of cells agglutination in groups treated with both CHL and BPDE was increased significantly; the colony formation efficiency in soft agar in groups treated with both CHL and BPDE (7.4 per thousand, 11.4 per thousand and 14.4 per thousand ) showed significant decrease (P < 0.05) in dose-dependent manner, as compared with that in group treated with BPDE alone (19.6 per thousand ). Cells treated with both CHL and BPDE or BPDE alone developed tumor in nude mice, a squamous carcinoma confirmed by histopathological examination. The volume of tumor in groups treated with both CHL and BPDE (0.43 +/- 0.13) cm(2), (0.22 +/- 0.04) cm(2) and (0.10 +/- 0.06) cm(3) was significantly smaller (P < 0.05) and dose-dependent, as compared with that in the group treated with BPDE alone (1.71 +/- 0.37) cm(3).

CONCLUSIONCHL showed significant antitransforming ability in human bronchial epithelial cell line induced by BPDE.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; toxicity ; Animals ; Anticarcinogenic Agents ; pharmacology ; Cell Transformation, Neoplastic ; drug effects ; Chlorophyllides ; pharmacology ; Mice ; Mice, Inbred BALB C ; Neoplasms, Experimental ; prevention & control

OBJECTIVETo investigate the effects of sodium copper chlorophyllin (SCC) on the proliferation, differentiation and immunomodulatory function of mesenchymal stem cells (MSCs) from mice with aplastic anemia.

METHODSA mouse model of aplastic anemia was established by exposure of BALB/c mice to sublethal doses of 5.0 Gy Co60 γ radiation, followed by transplantation of 2×10(6) lymph node cells from DBA/2 donor mice within 4 h after radiation. Aplastic anemic BALB/c mice were randomly divided into six groups: the treated groups, which received 25, 50, or 100 mg/kg/day SCC, respectively; a positive control group treated with cyclosporine A (CsA); and an untreated model control group (model group); while, the non-irradiated mice as the normal control group. SCC or CsA were administered by gastrogavage for 20 days, starting on day 4 after irradiation. Peripheral blood cells were counted and colony-forming fibroblasts (CFU-F) in the bone marrow were assayed. The ability of MSCs to form calcium nodes after culture in osteoinductive medium was also observed. The immunosuppressive effect of MSCs on T lymphocytes was analyzed by enzyme-linked immunosorbent assay and flow cytometry, to evaluate the efficacy of SCC in mice with aplastic anemia.

RESULTSPeripheral blood white cell and platelet counts were increased by medium and high SCC doses, compared with the untreated control. CFU-Fs were also increased compared with the untreated control, and the numbers of calcium nodes in MSCs in osteoinductive medium were elevated in response to SCC treatment. The percentage of Forkhead box protein 3 (FOXP3(+)) T cells was increased in T cell-MSC cocultures, and the cytokine transforming growth factor β1 was up-regulated in SCC-treated groups.

CONCLUSIONThe results of this study suggest that SCC not only promotes the proliferation and differentiation of MSCs, but also improves their immunoregulatory capacity in mice with aplastic anemia.

Anemia, Aplastic ; blood ; pathology ; therapy ; Animals ; Anthraquinones ; metabolism ; Biomarkers ; metabolism ; Bone Marrow Cells ; drug effects ; pathology ; Calcium ; metabolism ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Chlorophyllides ; pharmacology ; Colony-Forming Units Assay ; Female ; Immunosuppression ; Leukocyte Count ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred DBA ; Osteoblasts ; drug effects ; pathology ; Platelet Count ; T-Lymphocytes ; drug effects