ObjectiveBased on sequential metabolism and ultra-high performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）， to identify the prototype components and metabolites of Tongfengding capsules in rat plasma. MethodAn ACQUITY UPLC BEH Shield RP18 column （2.1 mm×100 mm， 1.7 µm） was used for gradient elution with 0.1% formic acid aqueous solution （A）-acetonitrile （B） as the mobile phase （0-1 min， 5%B； 1-2.4 min， 5%-10%B； 2.4-13.5 min， 10%-32%B； 13.5-18.5 min， 32%-90%B； 18.5-19 min， 90%-5%B； 19-21 min， 5%B） at a flow rate of 0.3 mL·min^-1， injection volume of 2 μL and column temperature at 35 ℃. The heated electrospray ionization （HESI） was applied under positive and negative ion modes， and the scanning range was m/z 100-1 500. By comparing the differences between the administered plasma and the blank plasma， the prototype components and metabolites in intestinal metabolism sample and liver metabolism sample prepared by intestinal perfusion with parallel blood collection， and comprehensive metabolism sample prepared by intragastric administration method were identified. ResultA total of 76， 53， 74 chemical components were detected in the intestinal metabolism sample， the liver metabolism sample and the comprehensive metabolism sample. A total of 100 components were identified from these different plasma samples， including 64 prototype components （34 alkaloids， 12 terpenoids， 9 organic acids， 6 flavonoids and 3 other components） and 36 metabolites. The main metabolic reactions involved in the formation of metabolites were glucuronidation， deglucosylation， dehydrogenation and hydroxylation. ConclusionThe chemical components of Tongfengding capsules can undergo a series of metabolic reactions in the intestine and liver， and a large number of metabolites are generated， among which alkaloids may be the leading component group for efficacy， which can lay the foundation for systematic elucidating the material basis of Tongfengding capsules in vivo.

Objective:To identify the chemical constituents of Fangfeng-Tongsheng Pills by ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-Q-Orbitrap HRMS). Methods:Take Agilent InfinityLab Poroshell 120 C18 column (150 mm×3.0 mm, 2.7 μm) and make water containing 0.1% formic acid and acetonitrile as gradient eluents. Data was collected with positive and negative ion modes using full scan and two-stage threshold-triggered mass modes (Full MS/dd-MS 2) and HESI source. The main chemical constituents of Fangfeng-Tongsheng Pills were identified according to the exact molecular mass, the cleavage fragments of MS 2, the retention time of reference substances, mzVault 2.0 MS database and literature review. Results:A total of 82 chemical constituents including 33 flavonoids, 13 chromones, 8 triterpenoid saponins, 6 coumarin, 5 iridoids, 4 phthalides, 4 terpenoids, 3 phenolic acids, 3 lignans, 2 anthraquinone and 1 alkaloids, were detected and identified.Conclusions:UPLC-Q-Orbitrap HRMS could identify the main chemical constituents of Fangfeng-Tongsheng Pills rapidly and accurately, which may provide a theoretical basis for further studies on the mechnism and quality control of this formula.

Objective:To analyze the metabolites of isoacteoside in rat's urine after oral administration by UHPLC-LTQ-Orbitrap and then summarize its metabolic pathways.Methods:The rats were randomly divided into treatment and control groups. Isoacteoside dissolved in saline was orally administered to the rats in the treatment group witht a single dose of 100 mg/kg. At the same time, saline was orally administered to the control group with the same volume. The urine samples were collected for 12 h and then purified. Sample analyses were performed on a Thermo Scientific BOS Hypersil C18 column (2.1 mm × 150 mm, 2.4 μm), the mobile phase consisted of water containing 0.1% formic acid-acetonitrile in a gradient program, the flow rate was set at 0.3 ml/min and the column was maintained at 30 ℃. The urine samples of the treatment group and control groups were detected with negative ion mode.Results:The metabolites were identified according to their protonated molecular ions and fragment ions and by comparing the mass data with that of reference standards and the published data. In total, 8 metabolites of isoacteoside were detected and identified in the urine samples of treatment group and the major metabolic pathway of isoacteoside included glucuronide conjugation, dehydroxylation, hydrolyzation, methyl conjugation and sulphate conjugation.Conclusions:UHPLC-LTQ-Orbitrap could be used to analyze the main metabolites and metabolic pathways of isophylloside in rats, which can provide references for further studies on pharmacodynamics and pharmacological mechanisms.

Objective DryLab software was used to assist high performance liquid chromatography (HPLC) method to test and isolate six Cytochrome P450 (CYP450) probe substrates.Methods Six CYP450 probe substrates were selected and the right HPLC method was developed and validated with the assistance of DryLab software.Results The new HPLC method with the assistance of DryLab software could test and isolate six probe substrates with degrees of isolation more than 2.00. The correlation coefficients (R> 0.999 8) indicated high linear correlation between the concentrations and the peak areas among six probe substrates. Recovery studies showed good results for all the probe substrat from 86.38% to 110.29%. And therelative standard deviation (RSD) ranged from 1.69% to 3.80% with its intra-day and inter-day precision ranging from 0.42% to 2.01%, and 1.36% to 2.29%, respectively.Conclusions The developed HPLC method with the assistance of DryLab could test and isolate six probe substrates with shortertime than the HPLC method alone.

BACKGROUND:Evaluation of vertical jumping ability is usual y only limited to height measurements. The measurements of parameters that describe kinetic factors may provide a better assessment of a patient’s jumping ability.
OBJECTIVE:To determine the deficit in one-legged vertical jumping ability and to clarify the relationships between the maximum jumping height and the maximum power, force and velocity during one-legged vertical jumps after anterior cruciate ligament reconstruction.
METHODS:Twenty-five healthy subjects (10 males and 15 females) and 25 anterior cruciate ligament reconstructed patients (10 males and 15 females) participated in this study. The isokinetic quadriceps femoris strength and one-legged vertical jumping ability were evaluated by the height, power, force and velocity in al subjects.
RESULTS AND CONCLUSION:(1) The maximum height of the one-legged vertical jumps was only significantly correlated with the maximum force in the healthy subjects (P<0.05). (2) However, for the reconstructed and unreconstructed legs in anterior cruciate ligament reconstructed patients, the maximum jumping height was significantly correlated with the maximum power, force and velocity during one-legged vertical jumps (P<0.05). (3) These findings suggest the importance of a knee strategy during one-legged vertical jumps for rehabilitation after anterior cruciate ligament reconstruction. Assessment of the jumping ability after anterior cruciate ligament reconstruction may be determined by the maximum power instead of the maximum jumping height.

Objective To study the multicomponent in vivo dynamic process in Chuanxiong Rhizoma;To elaborate in vivo metabolic profiling. Methods HPLC was used to establish the fingerprint of aqueous extract of Chuanxiong Rhizoma, and multicomponent changes were detected at the same time. Closed-loop intestine method was used to study the multicomponent changes of oral administration of Chuanxiong Rhizoma after stomach-intestine-liver process. Results Totally 17 components were detected in the fingerprint of aqueous extract of Chuanxiong Rhizoma and they were basically stable in the digestive juice. For in vivo metabolism, 4 components were metabolized by intestinal flora;3 components were metabolized by liver;2 new components were the metabolites of intestinal flora;1 component was the metabolite of liver. Conclusion Multicomponent sequential metabolism and closed-loop intestine method were used to clarify that multicomponent metabolic profiling was feasible, and it could provide experimental basis for the metabolism of traditional Chinese medicine.

Objective To investigate dynamic metabolism in vivo of Ginkgo Folium Tablet under the guidance of sequential metabolism thoughts. Methods In situ closed-loop in rats was carried out to study sequential metabolism of Ginkgo Folium Tablet through oral digestive system, namely to investigate and compare the intestinal flora metabolism, the gut wall metabolism and hepatic metabolism, combined with chromatographic fingerprint of blood samples. Results The analysis showed that 12 peaks in Ginkgo Folium Tablet were metabolized by intestinal flora, and 7 peaks generated through the gut wall. Most components of Ginkgo Folium Tablet were metabolized in liver, and 3 original medicine components were directly into the blood. Conclusion This study conducts a qualitative description of metabolism of Ginkgo Folium Tablet in different parts of the oral route, and provides references for the quality control, mechanism explanation and secondary development for Ginkgo Folium Tablet.

Objective To select the components for quality control of Fructus Lycii based on the absorption of its extract. Methods To investigate metabolism of components of Fructus Lycii, everted rat gut sacs was carried out as well as the blood was taken from abdominal aorta,.and all samples were analysised by HPLC. Results There are twelve constituents absorbed between ileum and jejunum of rat , and four constituents were detected in the blood. Compound 2, 3, 4, 5, 6, 9, 10, 11 were absorbed in prototype forms in the intestine directly,and compound 1, 7, 8, 12 were new ones. On the other hand, four compositions(3, 7, 10, 13)could be absorbed into blood through analysis serum samples obtaining from aorta abdominalis of rats. Two of them (3, 10)could be absorbed directly by intestine, while(7)was absorbed into blood in new form . Conclusion Based on the intestinal absorption experiment and analysion of compsition in blood, components (3, 10, 13) can be the quality control ingredients of Fructus Lycii.

This study was aimed to determine the solubility an d oil-water partition coefficient of main active com-ponents in Ge-Gen Qin-Lian (GGQL) Tablets (puerarin, baicalin and berberine hydrochloride) in phosphate buffer solution of different pH values and under the background of many components. Solubility of puerarin, baicalin and berberine hydrochloride in different medium pH, and oil-water partition coefficient of the octanol-water and oc-tanol-buffer system were determined by HPLC method. The results showed that the solubility and oil-water partition coefficient of puerarin, baicalin and berberine hydrochloride were varied with the change of pH, and varied under the background of components. At pH 7.4, the solubility was the biggest;puerarin was 7.56 mg·mL-1;baicalin was 17.07 mg·mL-1; berberine hydrochloride was 3.57 mg·mL-1. Oil-water partition coefficient P of these components at pH 1.0 was bigger;puerarin was 0.420 (lgP=-0.38);baicalin was 10.783 (lgP=1.03);berberine hydrochloride was 0.267 (lgP=-0.57). It was concluded that lipid solubility of puerarin, baicalin and berberine hydrochloride at pH 1.0 was better. It was speculated that better absorption in the stomach, and low lipid solubility under other pH. It was speculated that lipid solubility may be one of the reasons affecting the intestinal absorption.

Objective To research the metabolism and distribution ofsilybin in normal rats and liver injury model rats.Methods The normal rats group and immunity liver injury rat models were fed with the same dose ofsilybin capsule,and HPLC was used to determine the silybin concentration in biological samples in different time.Results The silybin concentration in the normal group in biological samples was higher than the model group at different time.In the normal group,the consequence of silybin concentration in each viscera distribution from top to bottom was liver＞kidneys＞plasma＞heart,while in the model group the fact was kidneys＞heart＞liver＞plasma.Conclusion The difference of metabolism and distribution of silybin in normal rats and liver damage model rats was obvious.