ABSTRACT
ObjectiveUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used to identify the metabolites of harmine in rats， in order to explore the differences in distribution of metabolites in rats after single dose（40 mg·kg-1） intragastric administration of harmine， as well to speculate the metabolic pathways. MethodSD rats were given a single dose of harmine by intragastric administration. Plasma， bile， urine and feces samples were collected after administration， and the samples were processed for determination by UPLC-Q-TOF-MS. The separation was performed on an ACQUITY UPLC™ HSS T3 columu（2.1 mm×100 mm， 1.8 μm） with acetonitrile（A）-0.1% formic acid aqueous solution（B） as mobile phase for gradient elution（0-2 min， 5%A； 2-9 min， 5%-35%A； 9-9.5 min， 35%-100%A； 9.5-12 min， 100%A； 12-12.5 min， 100%-5%A； 12.5-14 min， 5%A）， the mass spectra were obtained in positive ion mode with electrospray ionization（ESI）， the scanning range was m/z 50-1 200. The metabolites of harmine were identified based on the information of the obtained compounds and the literature data， and the metabolic pathways were hypothesized. ResultA total of 42 compounds（harmine and its metabolites） were identified in rats， including 27 in plasma， 17 in bile， 26 in urine and 13 in feces. The metabolic pathways involved in these 42 metabolites included monohydroxylation， dihydroxylation， demethylation， glucuronidation and sulfation. ConclusionHarmine can undergo phase Ⅰ and phase Ⅱ metabolic reactions in rats， and the prototype drug is metabolized rapidly in vivo， and the metabolites are mainly excreted by the kidneys， which can provide a reference basis for the pharmacodynamics and material basis of harmine.
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OBJECTIVE： To establish a method for the determination of harmine derivative DH-330 in rat plasma and to use it for pharmacokinetic behavior evaluation of DH-330 in rats after intragastric administration. METHODS： Using tinidazole as internal standard， after pre-treatment of acetonitrile precipitated protein， UPLC-MS method was adopted to determine the plasma concentration of DH-330. UPLC analysis was performed on Waters ACQUITY BEH C18 column （50 mm×2.1 mm，1.7 μm） with mobile phase consisted of acetonitrile-methanol-0.5％ formic acid aqueous solution（15 ∶ 55 ∶ 30， V/V/V） at flow rate of 0.4 mL/min， while the column temperature was 30 ℃， and sample size was 5 μL. MS analysis was conducted by electrospray ionization source， positive ion scanning， ion source temperature at 124 ℃， DH-330 detection of mass to charge ratio （m/z） of 335.8→334.8， and internal standard m/z of 247.0→81.0. Six Wistar rats were given DH-330 suspension（50 mg/kg） intragastrically. Blood samples were collected from fundus venous plexus capillary before administration （0 h） and 0.25，0.5，1，2，4，6，8，12，24 h after administration. Plasma concentration of DH-330 was determined and plasma concentration-time curves were drawn. Pharmacokinetic parameters were calculated by using Kinetica 5.0 software. RESULTS： The linear ranges of DH-330 were 25.05-2 004 ng/mL（r＝0.999 8），and the limits of quantitation was 25.05 ng/mL. RSDs of intra-day and inter-day were all less than 10％. The accuracy RE was －9.76％ to 4.55％. The extraction recovery was higher than 85％（RSD＜5％）. Stability RE was －2.53％ to 2.29％. They were not affected by matrix effect or residual effect of injection. The pharmacokinetic parameters of DH-330 in rats after intragastric administration included that cmax was （1 162.43±241.72）ng/mL，AUC0-∞ was （3 242.93±652.31）ng·h/mL，t1/2 was （1.93±0.61）h， MRT was （3.23±0.30）h，CL was （16.80±5.30）L/h·kg， Vss was （54.78±19.64）L/kg. CONCLUSIONS： The established method is simple， specific， sensitive， precise and recovery， which can be used for the plasma concentration determination of DH-330 in rats. DH-330 has short half-life， rapid absorption and large apparent distribution volume after intragastric administration in rats， which indicates that it has high lipophilicity and may be mainly distributed in tissues.
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OBJECTIVE:To prepare the Albendazole nanoliposomes freeze-dried power and study its properties. METHODS:Freeze-drying method was conducted to prepare Albendazole nanoliposomes freeze-dried power,using the particle size,encapsula-tion efficiency,appearance,redispersibility as indexes,single factor test was combined with orthogonal test to screen freeze-drying preparation technology. The morphological changes,particle size,Zeta potential,moisture content,12 months stability at 4 ℃ be-fore and after freeze-drying were detected. RESULTS:Plus a total content of freeze-dried protective agent was 10%,the ratio of glucose-trehalose-mannitol was 1.0:1.0:3.0,using quick-freeze,pre-freezing 18 h in -35 ℃ refrigerator,dry-freezing 48 h to ob-tain freeze-dried powder. Compared with before freeze-drying,the freeze-dried liposomal morphology had no obvious changes, showing clear phospholipid bilayer membrane structure;the particle sizes before and after freeze-drying were (208.63 ± 1.04) nm and (223.04 ± 2.02) nm,Zeta potentials were (-15.6 ± 0.04) mV and (-19.4 ± 0.06) mV,encapsulation efficiencies were (94.62±0.49)%and(91.10±0.46)%(n=3),respectively. Compared with liposomes,liposomes freeze-dried power had good sta-bility in 12 months at 4 ℃. CONCLUSIONS:Albendazole nanoliposomes freeze-dried power is prepared successfully,its stability is superior to albendazole nanoliposomes,and the freeze-drying technology is feasible.
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Objective To establish the quality standard for Complex Prescription Kudouzi Capsules.Methods TLC was employed to identify Crataegus pinnatifida Bge.、Cassia obtusifolia L.and sophora alopecuroides L..The content of chrysophanol in Cassia obtusifolia L.was determined by HPLC.Results Spots obtained from the test solutions had the same color in reference solution and medical material in the same location,and the blank solution had no interference.The linear range of chrysophanol was 0.028 4 ～ 0.908 8 ?g,r =0.999 7(n =6),and the average recovery was 99.97%.Conclusion The characteristic of identification by TLC was highly specific,HPLC is accurate and reproducible,and they can be used effectively for the quality control of Complex Prescription Kudouzi Capsules.