OBJECTIVETo determine the encapsulation efficiency of silybin liposomes.

METHODAfter Silybin liposomes solution was separated through sphadex G-100 column, eluent collected was determined by the first derivative UV spectrometry.

RESULTThe calibration curve of Silybin was linear in the range of 0.5-30.0 mg x L(-1) (r = 0.9998). The average recovery was 101.7%. The relative standard deviation was 1.8%. The results showed that the encapsulation efficiency ranged from 65.1%-83.0% when the drug/phospholipid ratio varied from 0.02-0.06. The concentration of SLB in liposomes prepared by optimum technique was 760.4 mg x L(-1) (n = 4).

CONCLUSIONThe method is accurate, simple and suitable for the quality control of Silybin liposomes.

Drug Delivery Systems ; Liposomes ; chemistry ; Milk Thistle ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Silymarin ; administration & dosage ; analysis ; isolation & purification ; Spectrophotometry, Ultraviolet ; Technology, Pharmaceutical ; methods

This research uses six Agrobacterium rhizogenes R1601, R15384, R1000, A4, R1025 and R1 to infect silymarin explants to induce hairy roots and silibin. All of the six A. rhizogenes can induce Silybum marianum to generate hairy roots and the A. rhizogene A4 shows comparatively high infection on the plant. This research determines the condition to induce silymarin hairy roots by the factors of infection time, pre-culturing, co-culturing and pH value. The fact that MS liquid medium fits the proliferation of silymarin hairy roots is determined. Through PCR molecular identification, it can be seen that the DNA plasmids in the A. rhizogenes are successfully integrated into the genome of transformed roots. Using liquid chromatography, it is determined that the silibin content in silymarin hairy roots is 2.5 times that in the plant In this research, the silymarin hairy roots culturing system is established, which lays a foundation for the study of culturing silymarin hairy roots and producing silibin.
Agrobacterium ; genetics ; physiology ; Cell Culture Techniques ; methods ; Milk Thistle ; chemistry ; genetics ; growth & development ; microbiology ; Plant Roots ; chemistry ; genetics ; growth & development ; microbiology ; Silymarin ; analysis ; Transformation, Genetic

OBJECTIVETo investigate lyophilization of SM-SLN.

METHODThe parameters of lyophilization process was optimized. In addition, the protective effect of various types and concentrations of cryoprotectants were tested by shape, colour and disparity.

RESULTThe mixture of 2% lactose and 2% glucose could better prevent nanoparticles from aggregating, the optimal lyophilization process was followed: precooled at -45 degrees C for 10 hr; primary drying at -25 degrees C for 5 hr; secondary drying at 10 degrees C for 3 hr; finally drying at 30 degrees C for 6 hr.

CONCLUSIONChanges in particle size distribution during lyophilization could be minimized by optimizing the parameters of the lyophilization process and adding supporting agent.

Drug Carriers ; chemistry ; Freeze Drying ; methods ; Glucose ; chemistry ; Lactose ; chemistry ; Lipids ; chemistry ; Milk Thistle ; chemistry ; Nanotechnology ; Particle Size ; Plants, Medicinal ; chemistry ; Silymarin ; administration & dosage ; chemistry ; isolation & purification ; Technology, Pharmaceutical ; methods

OBJECTIVETo observe the effects of silymarin on hepatic microsomal and mitochondrial membrane fluidity in mice.

METHODLiver microsomal and mitochondrial membranes were labled by ANS and DPH. Membrane fluorensent intensity (F), fluorensent polarization(P) and microviscosily(eta) of liver microsome and mitochondria were determined.

RESULTSil increased the external membrane fluidities of liver microsome and mitochondria, and decreased the internal membrane fluidities of liver microsome and mitochondria. Pretreatment with CCl4, the external membrane fluidity of liver microsome and mitochondria were increased, and the internal membrane fluidities of liver microsome and mitochondria were decreased. After given sil 140,280 mg.kg-1, the increased external membrane fluidities of liver microsome and mitochondria were lowered, and the decreased internal membrane fluidities of liver microsome and mitochondria were enhanced in a dose-dependent manner.

CONCLUSIONThe protective effects of sil on liver injury may be related to the recovery of the membrane fluidities of liver microsome and mitochondria.

Animals ; Carbon Tetrachloride Poisoning ; Liver Diseases ; etiology ; pathology ; Male ; Membrane Fluidity ; drug effects ; Mice ; Microsomes, Liver ; drug effects ; Milk Thistle ; chemistry ; Mitochondria, Liver ; drug effects ; Plants, Medicinal ; chemistry ; Protective Agents ; pharmacology ; Seeds ; chemistry ; Silymarin ; isolation & purification ; pharmacology

AIMTo study the preparation of silymarin proliposomes. To study its physicochemic properties, its pharmacokinetical characteristics and bioavailability in rats after oral administration.

METHODSSilymarin proliposomes were prepared by film-deposition on carriers. When the proliposomes were contacted with water to form liposome suspensions, the tests of physicochemical properties including encapsulation efficiency, particle size and stability of the formed liposome suspensions were determined by HPLC, laser-particle-sizer and etc. The concentrations of non-conjugated and overall silymarin in plasma of rats and their pharmacokinetic behaviors after oral administration were studied by RP-HPLC. The pharmacokinetic parameters were computed by software program 3P97.

RESULTSThe encapsulation efficiency of silymarin liposomes could be more than 90%, with an average particle size of about 238.8 nm and a very good stability. The high bioavailability of silymarin proliposomes could be gotten by oral administration.

CONCLUSIONCompared with silymarin, silymarin proliposome is a stable and easily industrialized preparation and did enchance the gastrointestinal absorption of silymarin.

Administration, Oral ; Animals ; Area Under Curve ; Biological Availability ; Drug Carriers ; Drug Stability ; Liposomes ; Male ; Milk Thistle ; chemistry ; Particle Size ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Silymarin ; administration & dosage ; blood ; chemistry ; pharmacokinetics ; Technology, Pharmaceutical ; methods

AIMTo investigate the intestinal absorption of silybin (SLB) in male rats.

METHODSSingle-pass intestinal perfusion (SPIP) technique was performed in each isolated region of the small intestine at a flow rate of 0.1 mL x min(-1). The samples of perfusate and portal plasma were collected at the designated periods of time after rat intestinal perfusion and analyzed for drug by HPLC.

RESULTSThe absorption rate constant (k(a)) and the effective permeability (P(eff)) of SLB at 190 microg x mL(-1) were determined for each segment. These data indicated the absorption rate were duodenum > jejunum > ileum > colon. SPIP was also performed in duodenum with three concentrations of SLB (80, 190 and 300 microg x mL(-1)). The concentration dependent changes of k(a) and P(eff) were evident in the duodenum perfusion of silybin. At silybin perfusate concentrations of 80 microg x mL(-1), k(a) and P(eff) were different from the values at either of the two higher concentrations (190 and 300 microg x mL(-1), P < 0.05). However there was no difference between 190 microg x mL(-1) and 300 microg x mL(-1) groups. The drug mass appearing in the plasma further indicated the absorption were duodenum > jejunum > ileum > colon.

CONCLUSIONSLB can be absorbed in whole intestinal sections. When the concentration raises to a certain level, the uptake of SLB will not increase.

Animals ; Colon ; metabolism ; Duodenum ; metabolism ; Ileum ; metabolism ; Intestinal Absorption ; Intestine, Small ; metabolism ; Jejunum ; metabolism ; Male ; Milk Thistle ; chemistry ; Perfusion ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Silymarin ; blood ; isolation & purification ; pharmacokinetics

OBJECTIVETo investigate effect of particle size on oral absorption of silymarin-loaded solid lipid nanoparuicles.

METHODSolid lipid nanoparticles (SLN) of various sizes (150 nm, 500 nm and 1000 nm) using Compritol 888 ATO as the material and silymarin (SM) as a model drug were prepared. Silybinin concentration in plasma of rats were determined by RP-HPLC with UV detector. The main pharmacokinetic parameters were calculated by 3p97.

RESULTResults showed that the AUC of 150 nm SLN was 2.08 fold that of 500 nm SLN and 2.54 fold of 1000 nm SLN treated orally to rats (P < 0.05). The oral bioavailability of 150 nm SLN was remarkably higher than the other two size SLN.

CONCLUSIONThis has important implications in designing of SM-SLN as a new oral drug delivery system.

Administration, Oral ; Animals ; Area Under Curve ; Biological Availability ; Drug Carriers ; Drug Delivery Systems ; Excipients ; Fatty Acids ; Female ; Male ; Milk Thistle ; chemistry ; Nanostructures ; Particle Size ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Silymarin ; administration & dosage ; isolation & purification ; pharmacokinetics

Fructus Arctii is a traditional Chinese medicine. The main counterfeit species are the seeds of Arctium tomentosum, Onopordum acanthium, Silybum marianum, Saussurea costus, Amorpha fruticosa. Traditional identification methods or molecular barcoding techniques can identify Fructus Arctii and its counterfeit species. However, the identification of the mixture of it and its spurious species is rarely reported. In this paper, we sequenced the ITS2 sequences of Fructus Arctii and 5 kinds of spurious species mix powder by high-throughput sequencing to identify the mixed powder species and providing new ideas for the identification of Fructus Arctii mix powder. The total DNA in mixed powder was extracted, and the ITS2 sequences in total DNA was amplified. Paired-end sequencing was performed on the DNA fragment of the community using the Illumina MiSeq platform. The sequence was analyzed by the software FLASH, QIIME and GraPhlAn etc. The results showed that the high quality ITS2 sequences of 39910 mix samples were obtained from the mixed samples, of which the total ITS2 sequence of the samples genus was 34 935. Phylogenetic analysis showed that the samples contained Fructus Arctii, A. tomentosum, O. acanthium, S. marianum, S. costus and A. fruticosa. Using ITS2 sequences as DNA barcodes, high-throughput sequencing technology can be used to detect the Fructus Arctii and its spurious specie in mixed powder, which can provide reference for the quality control, safe use of medicinal materials of Fructus Arctii and the identification of mixed powder of traditional Chinese medicine.
Arctium ; chemistry ; classification ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; Drugs, Chinese Herbal ; standards ; Fabaceae ; Fruit ; High-Throughput Nucleotide Sequencing ; Milk Thistle ; Onopordum ; Phylogeny ; Saussurea

An herbal extract mixture and yogurt added to the herbal extract mixture were tested for their protective and therapeutic effects on ethanol-induced liver injury. The herbal extract mixture, yogurt and commercial drugs were used for treatment for two weeks prior to administering a single oral dose of ethanol (3 g/kg body weight). The herbal extract mixture and yogurt added to the herbal extract mixture were found to provide protection against ethanolinduced toxicity comparable to the commercial drug treatment, according to the serum and histopathological analysis. It was also shown that co-treatment with herbal extract mixture and yogurt against a triple oral dose of ethanol (2 g/kg body weight, over one week) provided protection against ethanol toxicity. After the initial set of experiments, the herbal extract mixture and yogurt treatments were extended for three more weeks. When compared to the positive control, further treatment with both the herbal extract and yogurt significantly reduced liver injury and resulted in a lower grade of lipid deposition.
Alnus/*chemistry ; Animals ; Body Weight/drug effects ; Brassica napus/*chemistry ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Eating ; Ethanol/antagonists & inhibitors/*toxicity ; Fabaceae/*chemistry ; Fermentation ; Liver/pathology ; Male ; Milk Thistle/*chemistry ; Oryza sativa/*chemistry ; Phytotherapy ; Plant Extracts/*therapeutic use ; Rats ; Rats, Sprague-Dawley ; Yogurt

A new HPLC-UV technique for the separation and analysis of 10 monosaccharides achieved within 13.5 min using 1-phenyl-3-methyl-5-pyrazolone (PMP) as the labelling molecule of the reductive monosaccharides has been established by combining common high performance liquid chromatography-UV and C18 column. The established technique was applied to the quantification of the monosaccharide components in extract of Silybum marianum. The results showed that the tested 10 monosaccharides as PMP derivatives were baseline separated under the HPLC conditions proposed. It was confirmed that Silybum marianum extract was composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose and arabinose with the molar ratio of 0.66:0.84:0.58:1.0:1.6:0.69:2.7:4.8. Quantitative recoveries of the compositional monosaccharides separated from the extract were in the range of 92.4%-104.0%, and the RSD values fell within 0.68%-3.81%. The results demonstrated that the proposed HPLC method was simple, rapid, convenient, and precise, and it was applicable to the analysis of the compositional monosaccharides of Silybum marianum extract.
Antipyrine ; analogs & derivatives ; chemistry ; Arabinose ; analysis ; Chromatography, High Pressure Liquid ; methods ; Galactose ; analysis ; Glucose ; analysis ; Glucuronic Acid ; analysis ; Hexuronic Acids ; analysis ; Mannose ; analysis ; Milk Thistle ; chemistry ; Monosaccharides ; analysis ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; isolation & purification ; Quality Control ; Rhamnose ; analysis ; Seeds ; chemistry ; Spectrophotometry, Ultraviolet ; methods ; Xylose ; analysis