大药学背景下的药物化学创新教育

在科技迅猛发展的今天,药物化学已从一门包含化学和生物学知识的传统课程发展成为涉及多学科的复杂体系。结合当前的大药学背景,从教学内容、教学方式和课程体系三个方面对药物化学创新教育进行探索和实践。

药物研发链与专业课程链相融合的大药学拔尖人才培养模式

高校专业课程链的设计和实施与我国药物研发、生产过程脱节是造成目前"大药学"拔尖人才的培养无法与飞速发展的制药工业相匹配的主要原因。以打造高端人才、培养提升整体创新能力和改善人才结构及分布为重点的高等教育改革,是我国大药学人才培养与社会需求接轨的前提,而药物研发链和专业课程链相融合是中国大药学人才培养与社会需求接轨的有效途径。通过双链融合,可有效消除因专业课程和药物研发脱节导致的人才培养失衡,提升高等教育拔尖人才的培养质量,促进与社会需求的契合度。

产学研循环互动创新人才培养模式的研究与实践——以温州医学院药学教育为例

产学研结合教育是高等教育改革与发展的方向,是提升学生创新能力和实践能力的重要途径,是推动我国经济建设和可持续发展的战略性举措。我们前期积极吸收国内外先进教学经验,充分发挥温州医学院现有条件,初步建立了一套产学研循环互动的教学模式,即以教学实验区为载体、产学研循环互动的教学体系,尝试将产学研相结合的思想贯彻到药学人才培养的全过程,探讨了产学研合作教育对药学人才创新能力培养的有效性。

学科前沿进展融入天然药物化学课堂教学的思考

在本科生课程中导入天然药物化学国际前沿研究进展,多学科交叉融合,对于优化知识体系、提高教学质量、开阔学生视野、激发学习兴趣、培养创新意识、强化专业逻辑思维能力具有重要意义。教学团队梳理目前教材内容和教学中的不足,为天然药物化学前沿性内容融入课堂提供教学素材和方法,以培养面向新世纪的药学人才。

Determination of Plasma Concentration of Cinnamic Acid by High-Performance Liquid Chromatography and Its Pharmacokinetics in Rats after Oral Administration of Zi-Shen Pill

Aim To develop a simple and sensitive high-performance liquid chromatographicmethod for determination of plasma concentration of cinnamic acid and pharmacokinetic study in ratsafter a single oral dose of traditional Chinese medicinal preparation Zi-Shen pill. Method Plasmasamples were acidified with hydrochloric acid and extracted with ethyl acetate . Cinnamic acid wasdetermined by HPLC using a G_(18) column. A mobile phase ofmethanbl-acetonitrile-water-triethyl-amine (7:22:73 = 0.2, V/V), with the pH adjusted to 4.0 withphosphoric acid, and with a UV detector set at 340 nm. Results The standard curve was linear overthe range of 1.92- 192.0 μg·mL^(-1). The LLOQ was 1.92 μg·mL^(-1) . The RSDs of within-day andbetween-day precision were < 8%. The mean recovery was 82.0% . Conclusion After validation, themethpd has been used to investigate the pharmacokinetic profiles of the traditional Chinesemedicinal preparation Zi-Shen pill.

Study on Kinetic Characteristics of Alliinase

[Objective] The kinetic characteristics of alliinase was studied to select the optimum reaction performance. [Method] Alliinase activity was measured to analysis the influence of temperature, pH, substrate concentration and metal iron. [Result] Alliinase was an enzyme with thermal instability. Its optimum reaction temperature was 29℃ and pH value was 6.1. The Vmax was 0. 439 IU/mg and Km was 0.483 mmol/L by using natural extract as substrate. Alliinase activity was activated when the K^＋ , Mg^2＋ , Na^＋ and Cd^2＋ existed and alliinase activity was inhibited when Cu^2＋ existed. [Condusion] Results showed that the kinetic characteristics of alliinase supply the academic foundation for development and application of garlic medical products.

Pharmacokinetics of Recombinant E. coli L-asparaginase in Rats

The distribution of ^(125)I recombinant E. coli L-asparaginase in tissues ororgans and the excretion in urine, feces and bile were studied with in vivo radioactive tracertechnique. The amount of radioactivity excreted in urine, feces and bile within 24 h afterintravenous administration of ^(125)I recombinant E. col L-asparaginase to rats was 68.95% ,4.44%and 5.36% of the dose respectively. ^(125)I recombinant E. coli L-asparaginase in plasma samples wasdetermined. The levels of structural intact molecule in plasma samples were evaluated by SDS-PAGEand bio-imaging analyzer system. Pharmacokinetic parameters were assessed with a model-dependentmethod. The concentration-time curves of recombinant E. coli L-asparaginase after intravenousinjection at 1 250 IU·kg^(-1), 2 500, IU·kg^(-1), 5 000 IU·kg^(-1) to rats were consistent withthe two-compartment model. The first and terminal elimination t_(1/2) were 0.52 ~ 0.63 h and 2.39 ~2.76 h respectively. AUC was linearly related to the doses. The results of distribution in tissuesor organs and excretion in urine suggested that the metabolites of the enzyme were cleared bymechanisms of urinary excretion. Pharmacokinetics parameters of recombinant E. coli L- asparaginasein rats are warranted for the design of future clinical trials.

A Preliminary Study on Biological Activity of Wikstroemia chamaedaphne Meissn

[ Objective ] The insecticidal and antibacterial bioactivity of Wikstroemia chamaedaphne Meissn were screened and bioactive substances in it were separated and purified. [ Method] The Wikstroemia chamaedaphne Meissn was conducted ultrasonic extraction in petroleum ether, ethyl acetate and methanol. The insecticidal activity of Wikstroemia chamaedaphne Meissn to Mythimna separata walker and aphid were determined. The antibacterial activity of Wikstroemia chamaedaphne Meissn to Fusarium graminearu, Glomerella cingulata, F. oxysporium f. sp niveum, Alternaria solani and Fusarium oxysporium were also determined. The bioactivity-guided methods such as opencolumn chromatography and Pre-HPLC method were used to separate active components in petroleum ether extract from Wikstroemia chamaedaphne Meissn. [ Result] When the concentration was 500 mg/L, 3 kinds of extracts from Wikstroemia chamaedaphne Meissn didn＇ t show obvious antibacterial bioactivity to 5 kinds of test samples. When the concentration was 5%, petroleum ether extract show certain topical toxicity to aphids. The ethyl acetate extract showed certain antifeedant activity to 3^rd instar Larvae of Mythimna separata Walker. The fraction F4 of petroleum ether extract possessed highest topical toxicity to aphids and the lethality was 60.00%. [ Conclusion] Wikstroemia chamaedaphne Meissn contained many insecticidal constituents whose active parts and mechanism were needed further researches.

Chemical Constituents of Euphorbia ebracteolata

Aim To study the chemical constituents of Euphorbia ebracteolata Hayata. Methods Column chromatography was used in the isolation procedure, while the structures of isolated compounds were elucidated by spectral data. Results Six compounds were isolated and their structures were identified as baccatin (1), 3-acetyl-β-amyrin (2), 3,3′-diacetyl-4,4′-dimethoxy- 2,2′,6,6′-tetrah ydroxy diphenylmethane (3), 2,4-dihydroxy-6-methoxy-3-methyl acetophenone (4), β-sitosterol (5), and daucosterol (6). Conclusion Baccatin was obtained from Euphorbia ebracteolata for the first time.

Manipulation of enteric flora in ulcerative colitis

TO THE EDITORReviewing the available therapeutic options in the medical treatment of ulcerative colitis, Xu et al.[1], have omitted to mention an important aspect in the pharmacological management of the disease, namely the possibility to promote clinical and endoscopic improvement by manipulating the enteric flora.

Age-related symptom and life quality changes in women with irritable bowel syndrome

AIM:To explore age-related changes in symptoms and quality of life(QoL) of women with irritable bowel syndrome(IBS).METHODS:Two-hundred and fifty-four female adult outpatients with IBS attending the Department of Gastroenterology at the First Affiliated Hospital of Nanjing Medical University between January,2008 and October,2008 were approached.Patients with a history of abdominal surgery,mental illness or those who had recently taken psychotropic drugs were excluded.A physician obtained demographic and abdominal symptom data.All patients were asked to complete the Zung Self-Rated Anxiety and Depression Scale(SDS/SAS) and the IBS-specific QoL questionnaire.The patients were divided into six groups according to age,in 10-year increments:18-27 years,28-37 years,38-47 years,48-57 years,58-67 years and 68-75 years(maximum 75 years).Age-related differences of abdominal pain or discomfort were analyzed using ranksum tests.Differences in SDS/SAS and IBS-QoL scores between age groups were analyzed using one-way analysis of variance.Pearson's correlations evaluated potential associations between IBS symptoms,psychological factors and QoL in each age group.RESULTS:There were no differences in the distribution of IBS subtypes between age groups(χ2 = 20.516,P = 0.153).Differences in the severity of abdominal pain/discomfort with age were statistically significant(χ2 = 25.638,P < 0.001);patients aged 48-57 years,58-67 years or 68-75 years had milder abdominal pain/discomfort than those in the younger age groups.The severity of anxiety or depressive symptoms did not differ between age groups(SDS,χ2 = 390.845,P = 0.110;SAS,χ 2 = 360.071,P = 0.220).Differences of IBSQoL scores were statistically significant between age groups(χ2 = 1098.458,P = 0.011).The scores of patients in the 48-57-year group were lower than those in the 18-27-year and 28-37-year groups(48-57-year group vs 18-27-year group,74.88 ± 8.76 vs 79.76 ± 8.63,P = 0.021;48-57-year group vs 28-37-year group,74.88 ± 8.76 vs 79.04 ± 8.32,P = 0.014).The scores in the 68-75-year group were lower than those in the 18-27-year,28-37-year and 38-47-year groups(68-75-year group vs 18-27-year group,71.98 ± 9.83 vs 79.76 ± 8.63,P = 0.003;68-75-year group vs 28-37-year group,71.98 ± 9.83 vs 79.04 ± 8.32,P = 0.002;68-75-year group vs 38-47-year group,71.98 ± 9.83 vs 76.44 ± 8.15,P = 0.039).Anxiety and depression were negatively correlated with QoL in all age groups(SDS and QoL:18-27-year group,r =-0.562,P = 0.005;28-37-year group,r =-0.540,P < 0.001;38-47-year group,r =-0.775,P < 0.001;48-57-year group,r =-0.445,P = 0.001;58-67-year group,r =-0.692,P < 0.001;68-75-year group,r =-0.732,P < 0.001.SAS and QoL:18-27-year group,r =-0.600,P = 0.002;28-37-year group,r =-0.511,P < 0.001;38-47-year group,r =-0.675,P < 0.001;48-57-year group,r =-0.558,58-67-year group,P = 0.001;r =-0.588,P < 0.001;68-75-year group,r =-0.811,P < 0.001).A negative correlation between abdominal pain severity and QoL was found in patients aged more than 58 years(58-67-year group,r =-0.366,P = 0.017;68-75-year group,r =-0.448,P = 0.048),but not in younger patients(18-27-year group,r = 0.080,P = 0.716;28-37-year group,r =-0.063,P = 0.679;38-47-year group,r =-0.029,P = 0.812;48-57-year group,r =-0.022,P = 0.876).CONCLUSION:Factors affecting QoL should always be treated in IBS,especially emotional problems in young adults.Even mild abdominal pain should be controlled in elderly patients.

A pharmacodynamic model of portal hypertension in isolated perfused rat liver

AIM： To develop a pharmacodynamic model of porta hypertension from chronic hepatitis. METHODS： Pathological changes and collagen depositions were analyzed using morphometry to confirm CCI4-induced chronic hepatitis. At do, d28, ds6 and d84 of the process, the portal perfused velocities （μL/min） in isolated rat livers were exactly controlled with a quanti-fied pump. The pressure （mmHg） was monitored with a Physiological System. The geometric concentrations of phenylephrine or acetylcholine were added to a fixed volume （300 mL） of the circulating perfusate. The equation, the median effective concentration and its 95% confidence intervals of phenylephrine or acetyl- choline were regressed with Prism-4 software in non-linear fit and various slopes. In the isolated perfused rat livers with chronic hepatitis, both median effective concentrations were defined as the pharmacodynamic model of portal hypertension.CONCLUSION： A pharmacodynamic model of portal hypertension in isolated perfused rat livers with chronic hepatitis was defined as the median effective concen- trations of phenylephrine and acetylcholine.

Pharmacokinetics and Bioavailability of 2-Amino-6-Cyclopropylamino-9-(2,3 -Dideoxy-β-D-glyceropent-2-enofuranosyl) Purine (Cyclo-D4G) in Rats

AimTo characterize the pharmacokinetics of 2 -amino-6-cyclopropylamino-9-(2,3-dideoxy-β-D-glyceropent-2-enofuran osyl) purine (Cyclo-D4G) following intravenous administration and oral administ ration to rats. MethodsThe concentrations of Cyclo-D4G in rat (Sprague-Dawley male rats) plasma and urine were analyzed by high performance liquid chromatography (HPLC). ResultsFollowing intravenous adm inistration to rats, concentrations of Cyclo-D4G in plasma declined with a term inal phase half-life of 0 78±0 14 h (±s). Total clearance was 0 90±0 21 L·h -1 ·kg -1 . Renal excretion of unchanged Cyclo-D4G accounted for approximately 20% of total clearance. Steady state volume of distr ibution was 0 91±0 07 L·kg -1 . After oral administration to rats, conce ntrations of Cyclo-D4G in plasma declined with a terminal phase half-life of 0 83±0 13 h (±s). Total clearance was 3 81±2 03 L·h -1 ·kg -1 . Renal excretion of unchanged Cyclo-D4G accounted for approximat ely 9% of total clearance. Oral bioavailability of Cyclo-D4G in rat was 26 9%. ConclusionThe favorable pharmacokinetic profiles and lower to xicity provide support for further development of Cyclo-D4G clinical trials.

Pharmacokinetics and tissue behavior of enrofloxacin and its metabolite ciprofloxacin in turbot Scophthalmus maximus at two water temperatures

Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fish infections. However, studies concerning the pharmacokinetics of enrofloxacin in turbot are limited. In this study, the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin, were investigated in the turbot following intravenous and oral administration at 10 mg enrofloxacin/kg body weight, at 16℃ and 10℃ water temperatures. The concentrations of enrofloxacin and ciprofloxacin in the main tissues （plasma, muscle, liver and kidney） were detected by HPLC. The results show that the plasma concentration-time data for enrofloxacin were best described as a two-compartment open model after intravenous and oral administration. Three pharmacokinetic equations were established between the concentrations and temperatures. The kinetic profile of enrofloxacin was temperature dependent. The absorption half-life of enrofloxacin was 1.99 h and 2.17 h after oral administration, whereas the elimination half-life of the drug was 98.63 h and 136.59 h at 16℃ and 10℃, respectively. The peak concentration of enrofloxacin in plasma and tissues was higher at 16℃ than that at 10℃, and the peak plasma concentration time in the liver was the shortest at both temperatures among those of other tissues. The plasma ℃/MIC ratio varied between 11.08 and 5 540.00 at 16℃; and between 7.92 and 3 960.00 at 10℃. The AUC/MIC ratio was 467.82-280 690.00 at 16℃, and 359.48-215 690.00 at 10℃. These ratios indicate that it is possible to obtain therapeutic efficacy. Very low levels of ciprofloxacin were detected. The AUC ratios of ciprofloxacin and enrofloxacin in plasma suggest that plasma ciprofloxacin might play a minor role in enrofloxacin treatment for turbot.

Cortical stimulation for treatment of neurological disorders of hyperexcitability: a role of homeostatic plasticity

Hyperexcitability of neural network is a key neurophysiological mechanism in several neurological disorders including epilepsy, neuropathic pain, and tinnitus. Although standard paradigm of pharmacological management of them is to suppress this hyperexcitability, such as having been exemplified by the use of certain antiepileptic drugs, their frequent refractoriness to drug treatment suggests likely different pathophysiological mechanism. Because the pathogenesis in these disorders exhibits a transition from an initial activity loss after injury or sensory deprivation to subsequent hyperexcitability and paroxysmal discharges, this process can be regarded as a process of functional compensation similar to homeostatic plasticity regulation, in which a set level of activity in neural network is maintained after injury-induced activity loss through enhanced network excitability. Enhancing brain activity, such as cortical stimulation that is found to be effective in relieving symptoms of these disorders, may reduce such hyperexcitability through homeostatic plasticity mechanism. Here we review current evidence of homeostatic plasticity in the mechanism of acquired epilepsy, neuropathic pain, and tinnitus and the effects and mechanism of cortical stimulation. Establishing a role of homeostatic plasticity in these disorders may provide a theoretical basis on their pathogenesis as well as guide the development and application of therapeutic approaches through electrically or pharmacologically stimulating brain activity for treating these disorders.

Mechanism of volatile oil from Chuanxiong(Chuanxiong Rhizoma)-Suhexiang(Styrax)-Bingpian(Borneolum)in treating angina pectoris based on network pharmacology and its protective effects on myocardial damage in rats

Objective To explore the pharmacodynamic material basis and mechanism of action of volatile oil from Chuanxiong(Chuanxiong Rhizoma)-Suhexiang(Styrax)-Bingpian(Borneolum)(hereinafter referred to as C-S-B volatile oil)in treating angina pectoris based on network pharmacology and to detect its protective effects against rat myocardial damage.Methods Gas chromatography-mass spectrometry(GC-MS)was used to determine the constituents of volatile oils from Chuanxiong(Chuanxiong Rhizoma),Suhexiang(Styrax),and Bingpian(Borneolum),and the targets of the three main constituents were found predicted and screened using the PharmMapper server,and Gene Cards and Coo LGe N databases.The STRING database and Cytoscape software were used to draw the protein-protein interaction(PPI)network,RStudio software was used to analyze Gene Ontology(GO)and Kyoto Encyclopedia of Genome and Genome(KEGG)pathways,and Cytoscape software was used to construct the component-target-pathwaydisease network.The rat model of myocardial injury was established by intraperitoneal injection of a large dose of isoprenaline hydrochloride.After continuous intervention with C-S-B volatile oil for 14 d,the ejection fraction(EF)and short axis shortening rate(FS)of the left ventricle were detected.The indices of myocardial damage were detected after hematoxylin-eosin(HE)staining.Results Fifteen volatile oil components from the C-S-B formula were identified.There are 470 targets of these volatile oil components and 401 angina-related genes.There are 28 core targets,including CHRM4,ADRA1 A,FGFR1,CHRM2,CYP2 A6,CHRM5,CHRM1,CHRM3,HDAC2,and MPO,etc..The results of the KEGG analysis indicated that the C-S-B formula probably interferes with the following pathways:neuroactive ligand-receptor interactions,calcium signaling,cytochrome P450 metabolism of heteropoietin,among others.The results of animal experiments showed that the C-S-B formula essential oil could significantly improve the following myocardial indices in rats with myocardial injury:EF,FS,left ventricular end-systolic diameter(LVIDs),left ventricular end-diastolic diameter(LVIDd),and stroke volume(SV),and all the differences were statistically significant(P<0.01).Conclusion The mechanism of action of volatile oil components in the C-S-B formula in treating angina pectoris was analyzed using multi-component,multi-target and multi-pathway systems,which has laid a foundation for further revealing its mechanism of action.Animal experiments have shown that the volatile oil of the C-S-B formula can improve EF,FS,and other indices of myocardial damage in a rat model,thus relieving myocardial damage caused by heart hyperactivity,improving cardiac function,and protecting against myocardial damage.

Pharmacokinetics and Biodegradation of Chitosan in Rats

Chitosan, an excellent biomedical material, has received a widespread in vivo application. In contrast, its metabolism and distribution once being implanted were less documented. In this study, the pharmacokinetics and biodegradation of fluorescein isothiocyanate(FITC) labeled and muscle implantation administrated chitosan in rats were investigated with fluorescence spectrophotometry, histological assay and gel chromatography. After implantation, chitosan was degraded gradually during its distribution to diverse organs. Among the tested organs, liver and kidney were found to be the first two highest in chitosan content, which was followed by heart, brain and spleen. Urinary excretion was believed to be the major pathway of chitosan elimination, yet 80% of chitosan administered to rats was not trackable in their urine. This indicated that the majority of chitosan was degraded in tissues. In average, the molecular weight of the degradation products of chitosan in diverse organs and urine was found to be <65 k Da. This further confirmed the in vivo degradation of chitosan. Our findings provided new evidences for the intensive and safe application of chitosan as a biomedical material.

EFFECT OF ELECTROACUPUNCTURE OF SCALP-POINTS ON ABNORMAL DISCHARGES OF NEURONS AROUND THE CEREBRAL HEMORRHAGE FOCUS IN THE RAT

Objective: To study the mechanism of electroacupuncture (EA) of scalp-points for regulating abnormal discharges of neurons in different regions around the cerebral hemorrhage focus by using neuro-electrophysiological methods. Methods: 80 Wistar rats (anesthetized with 20% urethane 1 g/kg, i.p.) were randomly divided into normal, saline, model and EA groups, with 20 cases in each group. Cerebral hemorrhage model was established by intracerebral injection of the rat’s own arterial blood sample (40 uL). In rats of saline group, the same volume of saline was given for intracerebral injection. Extracellular electrical activity of neurons of the caudate nucleus and parafascicular nucleus and Tail flicking latency (TFL) were used as the indexes. “Baihui”(百会 GV 20) and “Taiyang”(太阳 EX-HN 5) were punctured from GV 20 towards EX-HN 5 with filiform needles and stimulated electrically with stimulating parameters of strength of 1 V, frequency of 15 Hz and duration of 15 min. Results: Compared with normal group, TFL values of model group and EA group increased significantly (P<0.01); and compared with model group, those of EA group decreased significantly (P<0.01), suggesting that the pain threshold increased significantly in cerebral hemorrhage rats while after acupuncture stimulation, it lowered strikingly. Compared with normal and saline groups, the latency values of the pain excitement and inhibitory responses of the cellular discharges of the caudate and parafascicular nuclei in model and EA groups increased significantly (P<0.05～0.01), while after EA, it recovered apparently (P<0.01), showing an apparent regulative effect of EA on the abnormal changes of discharges of neurons around the cerebral hemorrhage focus. Conclusion: Scalp-acupuncture possesses an apparent regulatory effect on the abnormal electrical activity of neurons around the cerebral hemorrhage focus which may favor the early recovery of functional activity of neurons near the focus tissues.

Development of a method for the quantitative determination of geniposide in rat plasma by liquid chromatography-tandem mass spectrometry with positive/negative ion-switching electrospray ionization and its application in a pharmacokinetic study in rats

Geniposide is a major bioactive constituent isolated from Gardeniajasminoides Ellis. To evaluate the pharmacokinetics of geniposide in pre-clinical studies, a rapid and specific liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was developed and validated. After simple protein precipitation, geniposide was analyzed on a DiamonsilR C18 column with a mobile phase of 10 mM ammonium acetate and methanol （20：80, v/v） at a flow rate of 0.6 mL/min. Detection was performed in ＂Truncated＂ multiple-reaction monitoring （MRM） mode with positive electrospray ionization （ESI） at m/z 411→411 for geniposide, and MRM mode with negative ESI ionization at m/z 415→295 for puerarin （internal standard, IS）. Linearity was established in the concentration range from 10.0 to 5000 ng/mL. The extraction recoveries ranged from 84.8% to 90.5% at concentrations of 10.0, 500 and 4.5x 103 ng/mL. The lower limit of quantification （LLOQ） was 10.0 ng/mL with 50 ~tL plasma. The validated method was successfully applied to the pharmacokinetic study of geniposide in rats at a dose of 200 mg/kg by oral administration.

Development and validation of an RP-HPLC-UV method for the determination of daphnoretin in rat plasma

A sensitive RP-HPLC-UV method has been developed and validated for the quantification of daphnoretin in rat plasma. Daphnoretin was extracted from rat plasma by protein precipitation and liquid-liquid extraction. Separation was performed on a Diamonsil C18 column （200 mm× 4.6 mm, 5 μm） with a mobile phase of methanol-20 mmol/L ammonium acetate （adjusted to pH 3.2 with acetic acid, 42：58, v/v） at a flow rate of 1.0 mL/min. The UV detector was set at 345 nm and column temperature was set at 40 ℃. The calibration curves were linear over the concentration range of 0.020-2.00 ~tg/mL, The lower limit of quantification （LLOQ） of daphnoretin in rat plasma was 0.020 μg/mL. The intra- and inter-day relative standard deviation （RSD） for measurement of quality control （QC） samples （0.050, 0.200 and 1.60 μg/mL） ranged from 5.0%-10.6%. Relative error （RE） was from ±（1.2%-2.5%）. The validated method was used successfully in a pharmacokinetic study of daphnoretin in rats after intraperitoneal injection.