Improving intestinal absorption and antibacterial effect of florfenicol via nanocrystallisation technology.

To study the effects of nanocrystallisation technology on the intestinal absorption properties and antibacterial activity of florfenicol (FF). The florfenicol nanocrystals (FF-NC) were prepared by wet grinding and spray drying. Additionally, changes in particle size, charge, morphology, and dissolution of FF-NC in the long-term stability were monitored by laser particle sizer, TEM, SEM, paddle method, and the structure of FF-NC powder was characterised by nuclear magnetic resonance (NMR) test. The antibacterial activity, intestinal absorption and intestinal histocompatibility of FF-NC were investigated by the stiletto, mini broth dilution susceptibility test, in situ single-pass intestinal perfusion (SPIP) and haematoxylin-eosin (H-E) staining. After 12 months of storage, the particle size and zeta potential of FF-NC were 280.43 ± 8.21 nm and -19.64 ± 3.45 mV, and the electron microscopy results showed that FF-NC were nearly circular with no adhesion between particles. In addition, the drug loading, encapsulation efficiency, and dissolution of FF-NC did not change significantly during storage. The inhibition zone of FF-NC against Escherichia coli and Staphylococcus aureus was 21.37 ± 1.70 mm and 25.17 ± 2.47 mm, respectively. Compared with the FF, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of FF-NC are reduced, and the absorption rate constant (Ka) and efficient permeability coefficient (Peff) of FF-NC in the three intestinal segments were increased by 1.28, 0.25, and 9.10 times and 0.59, 0.17, and 6.0 times, respectively. The results of tissue sections showed that FF-NC had little damage to the small intestinal. Nanocrystallisation technology is an effective method to increase the intestinal absorption and antibacterial activity of FF.

Increased bioaffinity and anti-inflammatory activity of florfenicol nanocrystals by wet grinding method.

Context: The main objective of the current study is to improve the water solubility of florfenicol (FF) and evaluate changes in its pharmacokinetics and anti-inflammatory activity.Materials and methods: Florfenicol nanocrystals (FF-NC) were prepared by wet grinding combined with spray drying. The characterisations, pharmacokinetics, and anti-inflammatory activity of FF-NC were evaluated.Results: The particle size, polydispersity index (PDI), and zeta potential of FF-NC were 276.4 ± 19.4 nm, 0.166 ± 0.011, and -18.66 ± 5.25 mV, respectively. Compared with FF, FF-NC showed a better dissolution rate in media at different pH. Pharmacokinetic experiments showed the area under the curve (AUC0-t), maximum concentration (Cmax), and mean residence time (MRT) of FF-NC were about 4.62-fold, 2.86-fold, and 1.68-fold higher compared with FF, respectively. In vitro anti-inflammatory experiments showed that FF inhibited the secretion of tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), and synthesis of NO in a dose-dependent manner, while FF-NC showed a stronger anti-inflammatory effect than FF under the same dose.Conclusion: FF-NC are an effective way to improve the bioaffinity and anti-inflammatory effects of FF.

Controlled Release of the Nimodipine-Loaded Self-Microemulsion Osmotic Pump Capsules: Development and Characterization.

The present study was intended to develop a controlled released osmotic pump capsule based on Nimodipine (NM)-loaded self-microemulsifying drug delivery systems (SMEDDSs) in order to improve the low oral bioavailability of NM. To optimize the NM-loaded SMEDDS composition, the experiments of NM solubility in different oils, the pseudo-ternary phase diagram experiments and the different drug loading experiments were conducted in the preliminary screening studies. Controlled release of NM required an osmotic pump capsule comprising a coated semi-permeable capsule shell, plasticizer, and pore-forming agent. NM release follows zero-order kinetics after oral administration. Polyethylene glycol content, used as a pore-forming agent, coating mass, and drug release orifice size were key factors affecting drug release behavior according to the single methods and were optimized through response surface methodology. The NM-loaded SMEDDS droplet size and the 1H NMR mass spectrogram of the novel capsule were determined. The droplet size of the reconstituted microemulsion was 39.9 nm and 1H NMR analysis showed NM dissolution in the microemulsion. The dissolution test performed on three batches of NM-SMEDDS capsules-prepared using optimal preparation methods-indicated the capsule to deliver a qualified drug delivery with a zero-order release rate. The results demonstrated that NM-loaded SMEDDSs were successfully developed and displayed a qualified release rate in vitro.

Preparation and pharmacokinetic study of fenofibrate cubic liquid crystalline.

An LCC delivery system for Fenofibrate (Fen) was developed to improve its poorly oral bioavailability. Fen-LCC preparation methods were screened, and the prepared Fen-LCC was then characterized by a polarizing microscope and transmission electron microscopy (TEM). The spray drying technique was selected to dry and solidify particles into powder. The in vitro release of Fen-LCC was measured and in vivo pharmacokinetic experiments were carried out on rats after oral administration. Particles prepared through the high-temperature input method exhibited structural characteristics of LCC, and re-dissolved particles maintained the same features. The LCC delivery system can significantly improve in vitro release outcomes. After oral administration, AUCs of the suspension and LCC systems were measured at 131.6853 µg⋅h/ml and 1435.72893 µg⋅h/ml, respectively. The spray drying process presented here better maintains cubic structures, and the LCC system significantly improves bioavailability levels.

Establishment of quantitative retention-activity model by optimized microemulsion liquid chromatography.

The effective permeability coefficient is of theoretical and practical importance in evaluation of the bioavailability of drug candidates. However, most methods currently used to measure this coefficient are expensive and time-consuming. In this paper, we addressed these problems by proposing a new measurement method which is based on the microemulsion liquid chromatography. First, the parallel artificial membrane permeability assays model was used to determine the effective permeability of drug so that quantitative retention-activity relationships could be established, which were used to optimize the microemulsion liquid chromatography. The most effective microemulsion system used a mobile phase of 6.0% (w/w) Brij35, 6.6% (w/w) butanol, 0.8% (w/w) octanol, and 86.6% (w/w) phosphate buffer (pH 7.4). Next, support vector machine and back-propagation neural networks are employed to develop a quantitative retention-activity relationships model associated with the optimal microemulsion system, and used to improve the prediction ability. Finally, an adequate correlation between experimental value and predicted value is computed to verify the performance of the optimal model. The results indicate that the microemulsion liquid chromatography can serve as a possible alternative to the PAMPA method for determination of high-throughput permeability and simulation of biological processes.

Simultaneous Determination of Hydrochlorothiazide and Losartan Potassium in Osmotic Pump Tablets by Microemulsion Liquid Chromatography.

A rapid and efficient oil-in-water microemulsion liquid chromatographic (MELC) method has been optimized and validated for the determination of hydrochlorothiazide (HCT) and losartan potassium (LOP) in osmotic pump tablets. Samples were injected into a C18 (150 mm × 4.6 mm ID, 5 µm particle size) analytical column, which was maintained at 30°C. The most effective MELC system had a mobile phase consisting of 95% (v/v) of 3.0% (w/w) SDS, 6.0% (w/w) n-butanol, 0.8% (w/w) n-octane, 90.2% (w/w) water and 5% (v/v) acetonitrile (pH 5). The flow rate was 1.0 mL min(-1) and UV detection was performed at 265 nm. Linearity ranged from 2.5 to 12.5 µg mL(-1) for HCT and 10.0-60.0 µg mL(-1) for LOP (r > 0.999 for both drugs). The proposed method was rapid, precise (RSDs < 1.4%) and accurate (98.9% recovery for HCT and 101% recovery for LOP). It is applicable to simultaneous determination of HCT and LOP in osmotic pump tablets.

Determination of the lipophilicity of Salvia miltiorrhiza Radix et Rhizoma (danshen root) ingredients by microemulsion liquid chromatography: optimization using cluster analysis and a linear solvation energy relationship-based method.

We evaluated 26 microemulsion liquid chromatography (MELC) systems for their potential as high-throughput screening platforms capable of modeling the partitioning behaviors of drug compounds in an n-octanol-water system, and for predicting the lipophilicity of those compounds (i.e. logP values). The MELC systems were compared by cluster analysis and a linear solvation energy relationship (LSER)-based method, and the optimal system was identified by comparing their Euclidean distances with the LSER coefficients. The most effective MELC system had a mobile phase consisting of 6.0% (w/w) Brij35 (a detergent), 6.6% (w/w) butanol, 0.8% (w/w) cyclohexane, 86.6% (w/w) buffer solution and 8 mm cetyltrimethyl ammonium bromide. The reliability of the established platform was confirmed by the agreement between the experimental data and the predicted values. The logP values of the ingredients of danshen root (Salvia miltiorrhiza Radix et Rhizoma) were then predicted. Copyright © 2015 John Wiley & Sons, Ltd.

Determination of drug lipophilicity by phosphatidylcholine-modified microemulsion high-performance liquid chromatography.

A new biomembrane-mimetic liquid chromatographic method using a C8 stationary phase and phosphatidylcholine-modified (PC-modified) microemulsion mobile phase was used to estimate unionized and ionized drugs lipophilicity expressed as an n-octanol/water partition coefficient (logP and logD). The introduction of PC into sodium dodecyl sulfate (SDS) microemulsion yielded a good correlation between logk and logD (R(2)=0.8). The optimal composition of the PC-modified microemulsion liquid chromatography (PC-modified MELC) mobile phase was 0.2% PC-3.0% SDS-6.0% n-butanol-0.8% ethyl acetate-90.0% water (pH 7.0) for neutral and ionized molecules. The interactions between the analytes and system described by this chromatographic method is more similar to biological membrane than the n-octanol/water partition system. The result in this paper suggests that PC-modified MELC can serve as a possible alternative to the shake-flask method for high-throughput unionized and ionized drugs lipophilicity determination and simulation of biological processes.

[Study on self-microemulsifying membrane controlled-release drop pill of hawthorn leaves flavonoids].

To prepare the hawthorn leaves flavonoids self-microemulsifying membrane controlled-release coated drop pill, and to study its release rate in vitro and pharmacokinetics study in vivo. In order to improve the dissolution of hawthorn leaves flavonoids, self-microemulsifying technology was used to prepare the hawthorn leaves flavonoids self-microemulsion. Hawthorn leaves flavonoids self-microemulsifying drop pill was prepared with the PEG 6000. Studies were made on the in vitro release of flavonoids from hawthorn leaves self-micro-emulsifying membrane-moderated coated drop pills and the in vivo pharmacokinetic in rats. The prescription of flavonoids from hawthorn leaves self-micro-emulsifying drop pills was 0.25 g of flavonoids from hawthorn leaves, 0.25 g of iodophenyl maleimide, 0.375 g of polyethylene glycol 400, 0.375 g of cremophor RH 40 and 2 g of polyethylene glycol 6000. The optimized prescription was 4 g of ethyl cellulose 20, 0.64 g of polyethylene glycol 400, 1.8 g of diethyl phthalate, and the weight of coating materials increased by 3.5%. Flavonoids from hawthorn leaves self-micro-emulsifying membrane-moderated coated drop pills complied with the design of sustained-release in 12 h in terms of in vitro release and in vivo pharmacokinetic parameters in rats, and its bioavailability was 2.47 times of quick-release drop pills. Slightly soluble flavonoids from hawthorn leaves could be made into sustained-release preparations by the self-micro-emulsifying and coating technology.

Optimization of liquid chromatographic method for the separation of nine hydrophilic and hydrophobic components in Salviae miltiorrhizae Radix et Rhizoma (Danshen) using microemulsion as eluent.

In this study, we have proposed and developed a novel, environmental-friendly and simple method for separation of nine hydrophilic and hydrophobic components in Danshen using microemulsion liquid chromatography. The proposed method was optimized via the preliminary screening experiment and the experimental design. The following factors were investigated in preliminary screening experiment: pH of mobile phase, column type, the nature of surfactant, the nature of oil phase and additives. In order to simultaneously optimize resolution and analysis time, the chromatographic optimization function (COF) was adopted to evaluate chromatograms. The central composite design (CCD) was used to create the matrix of experiments for mapping the chromatographic response surface. Finally, the COF values were fitted into a second order polynomial model and the response surface methodology (RSM) was employed to find the optimal eluent constituents. The reliability of the established model was confirmed by the good agreement obtained between experimental data and predictive values. Based on the results from the preliminary screening experiment and the CCD optimization, the optimal mobile phase was identified as a solution consisting of 6.68% (w/w) polyoxyethylene lauryl ether (Brij35), 0.84% (w/w) cyclohexane, 6.92% (w/w) n-butanol, 85.56% (w/w) phosphate buffer (pH 6.60) and 8mM cetyltrimethyl ammonium bromide (CTAB).

[Study on fragmentation of vitexin and isorhamnetin-3-O-beta-D-rutinoside using electrospray quadrupole time of flight mass spectrometry].

OBJECTIVE: To investigate the fragmentation pathway of vitexin and isorhamnetin-3-O-beta-D-rutinoside with CID-TOF-MS. METHOD: Equipped with an LC-MS was carried out using an ultra-performance liquid chromatography, electrospray ionization quadrupole collision-induced dissociation-TOF-MS. RESULT: ESI-MS spectrum showed [M-H]- base peak of m/z 431. 0958 and m/z 623.1566. The CID-MS of vitexin showed five basic fragment ions, three of which corresponded to the glucosyl ring fracture: m/z 353, 341 and 311; other two were benzyl ion m/z 283, aglycone ion m/z 269. In addition, two low abundance ions, namely, m/z 161 and m/z 117, generated by RDA cracking ions, were also characteristic ions. The CID-MS of isorhamnetin-3-O-beta-D-rutinoside showed six main characteristic fragments ions corresponding to the loss of rhamnosyl m/z 477 and the glycosyl ring fracture: m/z 387, 357 and 311, and aglycone ion m/z 315. In addition, B ring generated m/z 300 and m/z 271 and C ring generated m/z 243 and the RDA cleavage generated m/z 151 and m/z 125. CONCLUSION: Those fragment ions can be used to quickly identify vitexin and isorhamnetin-3-O-beta-D-rutinoside.

[Study on the enrichment and purification of total flavonoids in Microcos paniculata by macroporous adsorption resin].

OBJECTIVE: To establish an optimum enrichment and purification process of total flavonoids in Microcos paniculata by macroporous resins. METHODS: Five kinds of resins were compared and the best one was chosen. Then the parameters of the process were optimized by single factor tests, uniform design and statistical methods. RESULTS: DI01 was selected for its excellent adsorption and desorption properties, 70% ethanol was found to be the best elution solution. As far as the yield was considered, the best result was based on the followings: feed rate-1.0 BV/h, elution flow rate-2.0 BV/h, sample concentration-7.88 mg/mL, eluting agent amount-2.0 BV, pH value 4.8; then the yield reached 90.18% and the purity was 54.37%. If the purity was considered, the best parameters wereas follows: feed rate-1.0 BV/h, elution flow rate-2.0 BV/h, sample concentration-2.0 mg/mL, eluting agent amount-2.8 BV, pH value 7.8; then the purity reached 61.77% and the yield was 80.25%. CONCLUSION: The total flavonoids of Microcos paniculata can be effectively purificated and separated by D101 macroporous resin.

[A new monoterpene glycoside from Paeonia lactiflora Pall].

AIM: To study the chemical constituents of Paeonia lactiflora. METHODS: The constituents of P. lactiflora were separated by using various kinds of modern chromatography and was identified its structure on the basis of spectral analysis. RESULTS: A monoterpene glucoside named albiflorin R1 was isolated from the roots of Paeonia lactiflora Pall. In the structure of albiflorin R1, the aglycone connected with a glucose at its 2-OH while the hemiacetal hydroxyl in glucose moiety was free. CONCLUSION: Albiflorin R1 is a new monoterpene glycoside.