This study was aimed to establish the thin layer chromatography (TLC) identification and content determination of Tibetan medicineJi-Ni De-Xie (JNDX). TLC was used to identifyBerberis kansuensisSchneid,Terminalia chebula Retz.,Phyllanthus emblicaandCurcuma LongaL. The high performance liquid chromatography (HPLC) was used to determine the content of berberine hydrochloride. The results showed that the TLC identification had a good resolution with clear spots and no interference from the negative reference. The content of berberine hydrochloride showed a good linear relationship with the peak area in the range of 0.05-0.45 mg·mL-1. The regression equation wasY = 40 679X-130.56. The average recovery was 99.07%. It was concluded that the established method was easy to operate with good repeatability. It can effectively control the quality of Tibetan medicine JNDX.

OBJECTIVETo analyze chemical constituents of Sini San its migrating components in rat plasma and study its in vitro and in vivo material base using ultra-performance liquid chromatography coupled with photo-diode-array detector and tandem mass spetrometry (UPLC-PDA-MS/MS).

METHODACQUITY UPLC BEH C18 column (2.1 mm x 100 mm, 1.7 microm) was adopted, with gradient elution system of water containing 2 mmol x L(-1) ammonium acetate and acetonitrile at flow rate of 0.2 mL x min(-1). The column temperature was maintained at 35 degrees C. The mass spectra were obtained by electrospray ionization source operating in both positive and negative ion mode. Ions were scanned from the m/z 100 to 1 000, and the characteristic ions were schizolysised twice to obtain the secondary MS data.

RESULTTwenty chemical constituents were detected, including paeoniflorin, glycyrrhizic acid, saikosaponins a and naringin. In vivo, there were 8 ingredients directly absorbed into blood after the administration of Sini San decoction, such as paeoniflorin, naringin and hesperidin. Besides, 6 metabolites were also detected, involving glucuronides, sulfate and sulfoglucuronides.

CONCLUSIONIn vitro and in vivo chemical materials of Sini San decoction is analyzed by UPLC-PDA-MS/MS to reflect in vitro and in vivo material base of Sini San decoction in a comprehensive and rapid manner and provide basis for further study on efficacious material basis of Sini San decoction.

Animals ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Light ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

This paper was designed to study metabonomic characters of the osteoporosis induced by high dose of hydrocortisone and the protective effects of Drynariae Rhizoma, which can replenish the kidney and strengthen the bones. A urinary metabonomics method based on ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS/MS) was developed. Clear separation of healthy control group, model group and treatment group was achieved by using the principal components analysis (PCA) and 9 significantly changed metabolites were identified as potential biomarkers of osteoporosis. Compared with the health control group, the model group rats showed lower levels of creatinine, citric acid, azelaic acid, hippurate, tryptophan and indoxyl sulfate together with higher levels of phenylalanine, cresol sulfate and phenaceturic acid. These changes in urinary metabolites suggest that the disorders of amino acid metabolism, energy metabolism, gut microflora and anti-oxidative damage are related to osteoporosis induced by high dose of hydrocortisone and the potential effect of Drynariae Rhizoma on all the four metabolic pathways.
Animals ; Chromatography, High Pressure Liquid ; Male ; Metabolomics ; Osteoporosis ; prevention & control ; urine ; Plant Extracts ; pharmacology ; Polypodiaceae ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry

Objective: To establish a simple, reliable and reproducible animal model of neonatal hypoxic-ischemic encephalopathy (HIE) with similar clinical pathological process to neonates. Methods: Seven days after birth, 180 Sprague-Dawley (SD) rats were randomly divided into six groups: blank control group, experimental control group and four hypoxia groups (8, 10, 12 and 14 min hypoxia groups). Those in the experimental groups were locally anesthetized with 5% lidocaine to separate their tracheas through blunt dissection, followed by tracheal clamping with vascular clamp for 8, 10, 12 and 14 min, respectively. Rats in the experimental control group were only treated with blunt dissection of trachea. No intervention was given to the blank control group. Due to significant reduction in rat survival rate after 14 min of hypoxia, no further morphological or behavioral examination was performed in this group. Rat brain tissue sections were stained with hematoxylin-eosin (HE) 12 h after modeling. Three days after modeling, the rat brain was weighted and the apoptosis of neural cells was detected with terminal deoxynucleotidyl transferase(TdT) mediated dUTP nick end labeling (TUNEL). Morris water maze was used to screen cognitive impairment in these rats at the age of two months. One-way analysis of variance was used for statistical analysis. SNK test and Dunnett 's T3 test were performed to compare homogeneous and non-homogeneous data between groups. Results: Systemic cyanosis, loss of consciousness, paled body, urinary and fecal incontinence, twitching of the limbs and tail and other abnormal behavior were induced by hypoxia. Ischemic necrosis, bleeding, nucleus shrinkage in a large number of neurons and hyperchromatic nuclei were observed in the 8, 10 and 12 min hypoxia groups. Three days after modeling, brain weights of rats in the 8, 10 and 12 min hypoxia groups were lower than those of the blank control group and experimental control group [(1.16±0.07), (1.04±0.06), (0.97±0.12), (1.31±0.06) and (1.28±0.09) g, F=36.437, P<0.001]. However, the numbers of apoptotic cortical [(22.83±4.52), (30.25±3.02), (39.18±5.04), (7.96±2.24) and (8.86±2.49)/400 scope field, F=164.532, P<0.001] and hippocampal CA3 neurons of that three hypoxia groups were higher than those of the two control groups [(14.63±2.26), (20.25±3.02), (24.81±1.98), (4.75±2.66) and (6.67±1.78)/400 scope field, F=141.026, P<0.001]. At the age of two months, rats in the 8, 10 and 12 min hypoxia groups had longer escape latency [(17.99±6.48), (23.07±9.90), (38.94±32.46), (14.37±6.06) and (12.78±7.21) s, F=26.912, P<0.001] and fewer times of platform crossings than those in the control group and experimental control group [(5.00±1.41), (4.90±1.29), (3.75±1.83), (7.57±1.16) and (7.14±1.15) times, F=14.336, P<0.001]. Conclusions Pathological changes in brain tissues and behaviors of rats after modeling are in line with the characteristics of classic animal model of HIE and similar to the clinical pathology and physiology of HIE, and this could be a new, simple, reliable and reproducible animal model of HIE, being capable of controlling the duration of hypoxia accurately.

Beijing ; COVID-19 ; Contact Tracing ; Humans ; Risk Factors ; SARS-CoV-2