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A fast,reliable,and cost-effective liquid chromatography-tandem mass spectrometry method was established to determine the effects of the traditional Chinese medicine employed to treat coronavirus disease 2019,namely,Lianhua Qingwen granules,Huoxiang Zhengqi capsules,Jinhua Qinggan granules,Shufeng Jiedu capsules,and Angong Niuhuang pills,on the pharmacokinetics of lopinavir/ritonavir in rats.Blood samples were prepared using the protein precipitation method and atazanavir was selected as the internal standard(IS).Separation was performed on an Agilent ZORBAX eclipse plus C18(2.1 mm x 50 mm,1.8 μm)column using acetonitrile and water containing 0.1％formic acid as the mobile phase for gradient elution.The flow rate was 0.4 mL/min and the injection volume was 2 μL Agilent Jet Stream electrospray ionization was used for mass spectrometry detection under positive ion multiple reaction monitoring mode at a transition of m/z 629.3→447.3 for lopinavir,m/z 721.3→296.1 for rito-navir,and m/z 705.4→168.1 for the IS.The method showed good linearity in the concentration range of 25-2500 ng/mL(r=0.9981)for lopinavir and 5-500 ng/mL(r=0.9984)for ritonavir.The intra-day and inter-day precision and accuracy were both within±15％.Items,such as dilution reliability and residual effect,were also within the acceptable limits.The method was used to determine the effects of five types of traditional Chinese medicines on the pharmacokinetics of lopinavir/ritonavir in rats.The pharmaco-kinetic results showed that the half-life of ritonavir in the groups administered Lianhua Qingwen granules and Huoxiang Zhengqi capsules combined with lopinavir/ritonavir was prolonged by approx-imately 1.5-to 2-fold relative to that in the control group.Similarly,the pharmacokinetic parameters of lopinavir were altered.Overall,the results of this study offer important theoretical parameters for the effective clinical use of five types of traditional Chinese medicines combined with lopinavir/ritonavir to reduce the occurrence of clinical adverse reactions.
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OBJECTIVE To examine the reversal effect of desipramine (DMI) on resistance to temozolomide(TMZ) in U251/TR cells and explore its mechanism. METHODS U251/TR cells were exposed to DMI (20-80μmol · L-1) or TMZ (0.5-10 mmol · L-1) for 24 h, cell viability was determined by cell counting kit-8 assay with IC50 calculated. The cytotoxicity of U251/TR cells treated with TMZ (1 or 2 mmol·L-1) in combination with DMI (20, 30 or 40 μmol · L-1) for 24 h was detected using CCK-8 assay. Synergism between DMI and TMZ was analyzed by the JIN Zheng-jun method. Apoptosis of U251/TR cells induced by TMZ 1 mmol · L-1, DMI 30 μmol · L-1,or their combination was examined by Hoechst33258 stains and caspase 3 activity was detected by luminescence analysis. Expression of C/EBP homologous protein (CHOP) was measured using quantitative real-time PCR and Western blotting. The survival rate of U251/TR cells treated with TMZ 1 mmol·L-1 and/or DMI 30μmol·L-1 was also assessed after silencing CHOP expression by small interference RNA (siRNA). RESULTS DMI or TMZ alone inhibited the growth of U251/TR cells significantly in a concentration-dependent manner (r 2=0.983,0.982,P1.15), ie, compared with TMZ alone, TMZ (1 mmol·L-1) com?bined with DMI (30 μmol · L-1) produced significant nuclear fragmentation and condensation (P< 0.05). In addition, DMI and TMZ in combination activated caspase 3 activity in U251/TR cells (P<0.05). Knock?down of CHOP by specific siRNA attenuated the synergistic effect of DMI in the presence of TMZ, the survival rate of the combined drug group raised from 51.8%to 62.2%(P<0.05). CONCLUSION The results suggest that DMI reverse resistance of U251/TR cells to TMZ through activation of the CHOP-depend?ently apoptosis pathway.
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Objective To develop a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of forsklin in rat plasma.Methods After extraction with methyl tert-butyl ether,chromatographic separation was performed on a C18 column with the mobile phase consisting of water ( 0.1％ formic acid)-acetonitrile in a gradient elution mode.A tandem mass spectrometer equipped with electrospray ionization (ESI) source was used as detector in the positive ion mode.Quantification was performed using multiple reaction monitoring (MRM) with the precursor product combination ions of m/z 411→375.3 and 285→193 for forsklin and diazepam.Results Good linearity was obtained in the 0.5-1000 ng/ml range for the analyte and the analytical method was validated in terms of specificity,precision,accuracy,recovery,stability and matrix effect.These assays gave RSD values always lower than 14.4％ and RE values between -3.5 ％ and 3.8％.In addition,the specificity,extraction recovery,stability and matrix effect were satisfactory.Conclusion Due to its high sensitivity,specificity and simplicity,the method could be used for pharmacokinetic studies of forsklin.
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Objective To establish the method for content determination of total flavonoids and polysaccharides in Radix Codonopsis Lanceolatae from Mount Taishan.MethodsThe content of the total flavonoids was determined by UV spectrophotometry and the polysaccharide content was measured with the phenol-sulphuric acid method.ResultsThe optimal detective wavelength was 510nm for the total flavonoids.Satisfactroy linear relationship with the Absorbability was achieved when the mass concentration of rutin was within a range of 8～48?g/ml(r=0.9986).The average recovery rate was 99.31%(RSD=1.82%,n=5).The optimal detective wavelength was 490nm for the polysaccharides.There was good linear relationship between the Absorbability and the content of glucose in a range of 5～50?g/ml(r=0.9964) with an average recovery of 99.12%(RSD=1.49,n=4).ConclusionThis method is sensitive,simple and accurate,and it can be used for the quality control of Radix Codonopsis Lanceolatae from Mount Taishan.
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Objective To study the antioxidative effect of the ethanol extract (e-EXT) and aqueous extract (a-EXT) of Codonopsis lanceolata from Mount Taishan in mice. Method The mice were divided into six groups at random. After the mice were orally treated with 5% Tween-80 (solvent,20 mL/kg),Vitamin C (0.1 g/kg),e-EXT (10,20 g/kg) and a-EXT (10,20 g/kg) once daily for 14 days respectively,the blood,liver and brain were collected for determining SOD,GSH-Px,GSH and MDA levels. Results The SOD,GSH-Px and GSH levels were enhanced significantly,and the MDA levels were reduced markedly in serum,liver and brain homogenate of the mice treated with 20 g/kg e-EXT and a-EXT. In the serum and liver homogenate of 10 g/kg a-EXT group,the levels of SOD,GSH-Px and GSH were increased,the contents of MDA were decreased. Conclusion The ethanol extract and aqueous extract of Codonopsis lanceolata from Mount Taishan could enhance the antioxidative ability of the mice,especially the large-dose aqueous extract.
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Objective To observe the influence of the coumarins from Radix Angelicae pubescentis (TCA) on the hypnotic effects of pentobarbital sodium and barbital sodium. Methods Mice were randomly divided into solvent (5% Tween-80, 20 mL/kg), chloromycetin (25 mg/kg) and TCA (50, 100 mg/kg) groups, which were administered orally with equivalent volume. One hour later, all the animals were given pentobarbital sodium (40 mg/kg, ip) or barbital sodium (200 mg/kg, ip), and then the latent period and duration of hypnosis were observed. Results Hypnotic duration of pentobarbital sodium was prolonged by 249.8%, 171.1% and 78.7% in chloromycetin and two TCA groups respectively (P
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OBJECTIVE:To probe into the regular pattern,related factors and prognosis of hemolytic reactions induced by Puerarin injection.METHODS:A total of 44 hemolytic cases induced by Puerarin injection reported in 26 papers of domestic journals published since 1994 were analyzed.RESULTS:Of the 44 cases collected,31 were males and 13 females,with a mean age of(67.5?9.6) years.The hemolytic reactions were induced either by Puerarin injection or Puerarin for injection with a mean dose of(445.2?123.8)mg and a mean concentration of puerarin(in infusion) of(1.55?0.52) mg?mL~(-1).The hemolytic reactions occurred chiefly after administration of puerarin for 7 consecutive days(7.96?3.84 days on average) and appeared as acute hemolytic anemia.The puerarin specific antibody was noted in immunological test.10 patients including 9 males and 1 male died at a mean age of(72.1?5.7) years.CONCLUSION:The occurrence and turnover of Puerarin Injection - induced hemolytic reactions are related to many factors,to which great attention should be paid.