LC-MS based strategy for chemical profiling and quantification of dispensing granules of Ginkgo biloba seeds.

Ginkgo biloba seeds have been used as a traditional Chinese medicine for hundreds of years to treat diseases such as cervicitis, cough, asthma and other lung diseases. As a novel form, the dispensing granules (GSDG) of Ginkgo biloba seeds have been widely employed in clinic. However, its chemical profiling is not yet clear, which has restricted in-depth research in many fields. In this study, a high performance liquid chromatography coupled with quardrupole time-of-flight mass spectrometry method was used for the component characteration with the help of accurate molecular weights, fragmentation pathways, reported data, literatures and even some reference standards. Furthermore, in multiple-reaction monitoring mode, a high performance liquid chromatography coupled with quadrupole linear ion trap mass spectrometry method was developed and applied for simultaneous determination of the bioactive phytochemicals. As a result, a total of 56 components in GSDG were identified including 12 amino acids, 9 organic acids, 6 nucleosides and nucleobases, 6 flavonoids, 5 vitamins, 5 terpenoid lactones, 4 carbohydrates and 9 other compounds As for quantitative analysis, glutamic acid, asparatic acid, histidine, ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, eucomic acid, N-(N-glucopyranosyl)-indoleacetylaspartate and N-(N-glucopyranosyl)-indoleacetylglutamate were selected as the analytes for quanlity marker of GSDG. After necessary validation tests, the developed quantitative method was successfully put into use for 10 batches of GSDG. In all batches, N-(N-glucopyranosyl)-indoleacetylaspartate was the richest phytochemical with the amount of 17.3-25.7 mg/g while ginkgolide J (0.0197-0.0335 mg/g) was determined to be the poorest. The study is supposed to exhibit a comprehensive chemical profiling and to provide some strong basis for preparation technology, quality control and even for action mechanism of GSDG, this novel form of Chinese medicine.

Insight of Silkworm Pupa Oil Regulating Oxidative Stress and Lipid Metabolism in Caenorhabditis elegans.

Silkworm pupa oil (SPO) contains unsaturated fatty acids, tocopherols, and phytosterols, which can regulate serum total cholesterol or be used as an antioxidant. In this study, we investigated the impacts of SPO on the antioxidant stress and lipid metabolism of Caenorhabditis elegans. The lifespan of the C. elegans fed with different SPO concentrations was determined. The levels of endogenous reactive oxygen species (ROS) were analyzed with the fluorescent probe method. The activity of antioxidant enzymes and the content of malondialdehyde (MDA) were analyzed. The transcription level of specific mRNA was characterized with q-PCR. The survival time of the mutant strain under oxidative stress was determined by daf-2 (CB1370) mutant, sod-3 (GA186) mutant, and skn-1 (EU31) mutant. As for the lipid metabolism, the lipid accumulation was determined with an Oil-Red-O (ORO) staining. The transcription level of specific mRNA was determined by q-PCR. The results showed that the SPO feeding enhanced the activities of antioxidant enzyme by upregulating the expression of the genes skn-1, and sod-3 to decrease the production of ROS and MDA, which prolonged the life of nematodes treated with juglone. ORO staining analysis indicated the feeding of SPO decreased intestinal fat accumulation, downregulated expression of fat-5, fat-6, fat-7, and nhr-80, and upregulated age-1 and tph-1 expression. Conclusively, SPO enhanced the antioxidant capacity by regulating the skn-1 and sod-3 expression of antioxidant gene and reducing the fat accumulation by the insulin/IGF signaling pathway and nuclear hormone receptor nhr-80 signaling pathway of nematodes. This study provides new evidence for the antioxidant and lipid-lowering mechanisms of SPO in C. elegans.

Daphnoretin Arrests the Cell Cycle and Induces Apoptosis in Human Breast Cancer Cells.

Emerging evidence has shown that daphnoretin, one of the main active ingredients of Daphne giraldii Nitsche, processes antitumor activities in several tumor cells (e.g., colon cancer, lung cancer, cervical cancer, and osteosarcoma). However, the antitumor effect and its mechanism in breast cancer are unexplored. In this study, our data indicated that daphnoretin obviously suppressed the proliferation of breast cancer MCF-7 and MDA-MB-231 cells. Further studies showed that daphnoretin remarkably increased the p21 level, decreased cyclin E and CDK2 levels, and then arrested the cell cycle at the S phase. Moreover, daphnoretin obviously lowered the BCL-2 level and raised the levels of BAX and cleaved caspase-9 and -3, leading to cell apoptosis. Furthermore, daphnoretin remarkably decreased the ratio of p-PI3K/PI3K and p-AKT/AKT in breast cancer cells. Collectively, these findings demonstrated that daphnoretin could suppress breast cancer cell proliferation through cell cycle arrest and inducing apoptosis, which is related to the PI3K/AKT pathway.

Novel maturity parameters for mature to over-mature source rocks and oils based on the distribution of phenanthrene series compounds.

Pyrolysis experiments of a low-mature bitumen sample originated from Cambrian was conducted in gold capsules. Abundance and distribution of phenanthrene series compounds in pyrolysis products were measured by GC-MS to investigate their changes with thermal maturity. Several maturity parameters based on the distribution of phenanthrene series compounds have been discussed. The results indicate that the distribution changes of phenanthrene series compounds are complex, and cannot be explained by individual reaction process during thermal evolution. The dealkylation cannot explain the increase of phenanthrene within the EasyRo range of 0.9% ∼ 2.1%. Adding of phenanthrene into maturity parameters based on the methylphenanthrene isomerization is unreasonable, even though MPI 1 and MPI 2 could be used to some extent. Two additional novel and an optimized maturation parameters based on the distribution of phenanthrene series compounds are proposed and their relationships to EasyRo% (x) are established: log(MPs/P) = 0.19x + 0.08 (0.9% < EasyRo% < 2.1%); log(MPs/P) = 0.64x - 0.86 (2.1% < EasyRo% < 3.4%); log(DMPs/TMPs) = 0.71x - 0.55 (0.9% < EasyRo% < 3.4%); log(MTR) = 0.84x - 0.75 (0.9% < EasyRo% < 3.4%). These significant positive correlations are strong argument for using log(MPs/P), log(DMPs/TMPs) and log(MTR) as maturity parameters, especially for mature to over-mature source rocks.