Identification of chemical constituents in Huangdi Anxiao Capsules by UPLC-Q-TOF-MS~E combined with UNIFI software / 中国中药杂志

In order to clarify the main chemical constituents of Huangdi Anxiao Capsules, an ultra-high performance liquid coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS~E) combined with Waters UNIFI software were successfully used to rapidly identify the chemical constituents in Huangdi Anxiao Capsules. The mass spectrometry data of chemical constituents from Huangdi Anxiao Capsules were collected by UPLC-Q-TOF-MS~E, and their structures were identified by the results of UNIFI software according to relative retention time of reference standards, MS feature fragments and literature data of each compound. A total of 100 compounds in Huangdi Anxiao Capsules were identified, including 25 compounds from Pueraria Lobate Radix, 22 compounds from Coptis Rhizoma, 6 compounds from Ophiopogonis Radix, 14 compounds from Eriobotryae Folium, 22 compounds from Rehmanniae Radix, and 15 compounds from Notoginseng Radix et Rhizoma. Among them, 3 compounds were common components. These 100 compounds included flavonoids, alkaloids, saponins and organic acids. This study systematically analyzed the chemical composition of Huangdi Anxiao Capsules, so as to provide evidences for defining the chemical material basis of Huangdi Anxiao Capsules.

Effect of Huangdi Anxiao Capsules on zebrafish vascular lesions induced by high glucose and high fat / 中国中药杂志

Zebrafish of different strains with 5 dpf (5 days post-fertilization) were selected and fed with 0.2% high-fat diet for 8 h and 3% glucose solution for 16 halternatively during the day and night for 4 consecutive days. The zebrafish model was established and randomly divided into model group, Huangdi Anxiao Capsules (260 mg·L⁻¹) group and pioglitazone (32 mg·L⁻¹) group. The drug treatment groups were given the water-soluble drugs, with a volume of 25 mL, and incubated in a 28 °C incubator for 4 days. To detect the exposure to the corresponding drugs, the normal control group was set up. Thirty zebrafish were included in each group. The effect of Huangdi Anxiao Capsules on vascular wall thickness, fluorescence intensity of islet beta cells, fluorescence intensity of macrophages, and blood flow velocity of zebrafish were detected. The expressions of vascular endothelial growth factor (vegfaa) and angiotensin converting enzyme (ACE) were detected by RT-PCR. The results showed that compared with the model group, Huangdi Anxiao Capsules can significantly reduce the thickness of the blood vessel wall, increase the fluorescence intensity of islet β cells and macrophages, increase the blood flow velocity in vivo, and decrease the ACE and vegfaa expressions in zebrafish. It is suggested that Huangdi Anxiao Capsules may alleviate zebrafish vascular lesions by regulating the expressions of ACE and vegfaa.

Serum fingerprint of drug-couple Psoralea corylifolia-Myristica fragrants / 中草药

Objective: To establish the chromatographic fingerprint of components of drug-couple Psoralea corylifolia-Myristica fragrants (PC-MF) absorbed into blood by UPLC. Methods: Twenty SD rats were administered with 10 different batches of PC-MF respectively. The UPLC profiles of serum samples were collected after treatment of PC-MF, blank serum samples and methanol extract were compared, and the transitional constituents to blood and their metabolites were analyzed. Results: The fingerprint of PC-MF was established. Thirteen common peaks were identified and the similarities were over 0.90. The 10 peaks from the in vitro test were prototype components in the product, and three peaks were metabolites. Conclusion: The serum fingerprint of PC-MF is established for the first time. It reflects the oral administration absorption into the blood and provides some data on pharamacodynamic basis study in vivo for PC-MF.

Determination and pharmacokinetics of main components for Psoralea corylifolia-Myristica fragrants drug pair by using UPLC-MS/MS / 中国中药杂志

To conduct multiple-reaction monitoring(MRM) quantitative analysis with ultra-high performance liquid chromatography coupled with mass spectrometry method(UPLC-MS/MS), determine the concentrations of psoralen, isopsoralen, bakuchiol and dehydrodiisoeugenol in plasma under positive iron mode with chloramghenicol as internal standard, and investigate the pharmacokinetics process of the main components before and after oral administration of drug pair Psoralea corylifolia -Myristica fragrants. Thirty-six SD rats were randomly divided into three group(A, B, C) and received P. corylifolia extract, P. corylifolia-M. fragrants extract, and M. fragrants extract respectively by intragastric administration. The plasma samples were collected at different time points. In the plasma samples, psoralen, isopsoralen, bakuchiol and dehydrodiisoeugenol showed good linear relationship within concentration rages of 0.098 125 to 39.25, 0.084 37 to 33.75, 0.046 875 to 18.75, and 0.11 to 2.2 mg•L⁻¹ respectively. The precision and stability results showed that the determination method of plasma concentration for such compositions was stable and reliable. The pharmacokinetic parameters obtained by DAS 2.0 showed varying differences before and after compatibility. According to the experimental results, the compatibility of P. corylifolia and M. fragrants can significantly impact the pharmacokinetic process of main components, expand their distribution and accelerate their metabolism and elimination in vivo.

Urine metabonomic study on Danzhi Jiangtang Capsule in treatment of type 2 diabetic rats based on UPLC/QTOF-MS / 中草药

Objective: To observe the metabolic profile changes caused by Danzhi Jiangtang Capsule (DJC) in treatment of type 2 diabetic rats and to study the possible metabolism. Methods: All the SD rats involved in the experiment were randomly divided into normal control, model, and DJC groups. The rats in the model and DJC groups were fed on high-fat diet for 4 weeks after ip injection with a small dose of streptozotocin to copy type 2 diabetic rat model. After the modeling, the rats in the DJC group were ig administered with DJC (1.26 g/kg) for 4 weeks, and the blood glucose was monitored weekly. After the treatment, urine of rats was collected, then all the rats were sacrificed, abdominal aortic blood was taken to detect the levels of insulin (INS) and glycated hemoglobin (GHb) in plasma. Using UPLC/QTOF-MS with principal component analysis (PCA) to analyze the metabolic profile changes, the metabolites were identified by databases, and metabolism pathways were analyzed. Results: Blood glucose monitoring and INS, GHb testing results showed that DJC could lower blood sugar and elevate plasma INS level. In the positive and negative ion modes, we screened seven and six variability markers respectively. The variability markers were related to glucose metabolism, tyrosine metabolism tryptophan metabolism, and oxidative stress-related reactions. Conclusion: DJC can lower blood sugar, repair islet cells to cure the type 2 diabetes, and the possible mechanism may relate to the regulation of energy metabolism, tyrosine metabolism, tryptophan metabolism, and oxidative stress.

Optimization of processing technology for xanthii fructus by UPLC fingerprint technique and contents of toxicity ingredient / 中国中药杂志

The experiment's aim was to optimize the processing technology of Xanthii Fructus which through comparing the difference of UPLC fingerprint and contents of toxicity ingredient in water extract of 16 batches of processed sample. The determination condition of UPLC chromatographic and contents of toxicity ingredient were as follows. UPLC chromatographic: ACQUITY BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acidwater in gradient mode, the flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 327 nm. Contents of toxicity ingredient: Agilent TC-C18 column (4.6 mm x 250 mm, 5 microm), mobile phase was methanol-0.01 mol x L(-1) sodium dihydrogen phosphate (35: 65), flow rate was 1.0 mL x min(-1), and detection wavelength was 203 nm. The chromatographic fingerprints 16 batches of samples were analyzed in using the similarity evaluation system of chromatographic, fingerprint of traditional Chinese medicine, SPSS16.0 and SIMCA13.0 software, respectively. The similarity degrees of the 16 batches samples were more than 0.97, all the samples were classified into four categories, and the PCA showed that the peak area of chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid were significantly effect index in fingerprint of processed Xanthii Fructus sample. The outcome of determination showed that the toxicity ingredient contents of all samples reduced significantly after processing. This method can be used in optimizing the processing technology of Xanthii Fructus.