Stability and drug dissolution evaluation of Qingkailing soft/hard capsules based on multi-component quantification and fingerprint pattern statistical analysis

Purpose: To carry out a post-marketing evaluation of the stability and drug dissolution of Qingkailing soft/hard capsules.Methods: High performance liquid chromatography with diode array detection (HPLC-DAD) method was developed for the determination of three key ingredients (chlorogenic acid, geniposide and baicalin) and fingerprints of QKL soft/hard capsules. Stability tests were carried out based on long-term testing. The drug release profile of Qingkailing soft and hard capsules were studied using semi-bionic incubation experiments.Results: The linearity, precision, stability, repeatability and recovery of HPLC and fingerprint all met the requirements of CFDA. Stability data from long-term studies showed that within 6 months the contents of the three key ingredients in both soft and hard capsules remained > 90 %. However, fingerprint pattern statistical analysis showed that the soft capsule is more stable than the hard capsule. Furthermore, the key ingredients of the hard capsule dissolved much faster (p < 0.05) than from the soft capsule. The level of dissolved drug of hard capsule is about 4 times the rate of soft capsule, after a 4-h incubation in gastric lavage fluid. In intestinal lavage fluid, more than 90 % of chlorogenic acid, geniposide and baicalin of hard capsule were dissolved in 2 h, while the soft capsule displayed a 12 h sustained release. Fingerprint pattern statistical analysis also showed that most of the components of soft capsule dissolved after 8 h.Conclusion: Compared with the hard capsule, Qingkailing soft capsule has certain advantages in stability and drug dissolution, which may affect the biopharmaceutics and the clinical effects of the drug.Keywords: Qingkailing capsule, Chlorogenic acid, Geniposide, Baicalin, Fingerprint, Sustained release, Principal component analysi

In Situ and in Vivo Study of Nasal Absorption of Paeonol in Rats

The objective of this work was to study the in situ and in vivo nasal absorption of paeonol. A novel single pass in situ nasal perfusion technique was applied to examine the rate and extent of nasal absorption of paeonol by rats. Various experimental conditions, such as perfusion rate, pH, osmotic pressure and drug concentration, were investigated. The in situ experiments showed that the nasal absorption of paeonol was not dependent on drug concentration, and fitted a first order process. The absorption rate constant, Ka, increased with an increase in perfusion speed. Paeonol was better absorbed in acidic solutions than in neutral or alkaline solutions. The value of Ka was higher in a hypertonic environment than under isotonic or hypotonic conditions. In vivo studies of paeonol absorption were carried out in rats and the pharmacokinetics parameters of intranasal (i.n.) and intragastric (i.g.) administration were compared with intravenous (i.v.) administration. The bioavailabilities of paeonol were 52.37% and 15.81% for i.n. and i.g, respectively, while Tmax values were 3.05 ± 1.46 min and 6.30 ± 0.70 min. MRT (Mean Residence Time) were 23.19 ± 6.46 min, 41.49 ± 2.96 min and 23.09 ± 5.88 min for i.n., i.g. and i.v. methods, respectively. The results demonstrate that paeonol could be absorbed promptly and thoroughly by i.n. administration in rats

Spread of Streptococcus suis Sequence Type 7, China

Streptococcus suis sequence type (ST) 7 has been spreading throughout China. To determine events associated with its emergence, we tested 114 isolates. In all 106 ST7 strains responsible for human outbreaks and sporadic infections, the tetracycline-resistance gene, tetM, was detected on the conjugative transposon Tn916. Horizontal transmission of tetM is suspected

Streptococcus suis Sequence Type 7 Outbreak, Sichuan, China

An outbreak of Streptococcus suis serotype 2 emerged in the summer of 2005 in Sichuan Province, and sporadic infections occurred in 4 additional provinces of China. In total, 99 S. suis strains were isolated and analyzed in this study: 88 isolates from human patients and 11 from diseased pigs. We defined 98 of 99 isolates as pulse type I by using pulsed-field gel electrophoresis analysis of SmaI-digested chromosomal DNA. Furthermore, multilocus sequence typing classified 97 of 98 members of the pulse type I in the same sequence type (ST), ST-7. Isolates of ST-7 were more toxic to peripheral blood mononuclear cells than ST-1 strains. S. suis ST-7, the causative agent, was a single-locus variant of ST-1 with increased virulence. These findings strongly suggest that ST-7 is an emerging, highly virulent S. suis clone that caused the largest S. suis outbreak ever described

Identification and molecular cloning of novel antimicrobial peptides from skin secretions of Odorrana versabilis and Rana palustris

Objective: Amphibian skin secretions are an abundant source of bioactive peptides, some of which could be developed as candidate drugs. Among these natural peptides, cytolytic peptides have attracted the most attention given that they might replace conventional antibiotics and help deal with the problem of microbial resistance. This study discovered two bioactive peptides, Brevinin-1-PLr and Nigrocin-2-OV, from two species frogs, the Chinese bamboo leaf odorous frog (Odorrana versabilis) and the North American pickerel frog (Rana palustris), respectively. Their antimicrobial, anticancer and hemolytic activities were also investigated. Methods: cDNA sequences encoding peptides were cloned from cDNA libraries constructed from the lyophilized secretions of the Chinese bamboo leaf odorous frog and the North American pickerel frog. By reversed-phase HPLC and MS/MS fragmentation sequencing, the encoded novel peptides, named Nigrocin-2-OV and Brevinin-1-PLr, were identified in skin secretions and their structures were confirmed. Replicates of both peptides were produced through solid phase peptide synthesis. Their antimicrobial and anticancer activity was studied against three types of microorganisms (Staphylococcus aureus, Candida albicans, and Escherichia coli) and five cancer cell lines (NCI-H157, PC-3, MDA-MB-435s, MCF-7, and U251MG). Their hemolytic activity was investigated using whole horse blood. Results: In this research, cDNA sequences encoding two novel 24-mer peptides were cloned from cDNA libraries constructed from the lyophilized skin secretions of the Chinese bamboo leaf odorous frog and the North American pickerel frog. Both of the peptides had the strongest inhibitory effect against C. albicans, and IC50 values against five cancer cell lines were all under 6 μM. Conclusions: Nigrocin-2-OV and Brevinin-1-PLr had the strong ability to inhibit the proliferation of studied microorganisms and tumor cell lines, with slight hemolytic activity. Compared with Brevinin-1-PLr, Nigrocin-2-OV exhibited higher antimicrobial and anticancer activity but slightly higher hemolytic activity

Investigation of Factors Affecting Microdialysis Probe Delivery and Recovery of Puerarin In Vitro

Purpose: To investigate in vitro the factors affecting microdialysis probe delivery and recovery of puerarin . Methods: The recovery and delivery of puerarin were tested for extraction efficiency and retro-dialysis methods. Factors such as drug concentration, stirring speed, additives and length of membrane were studied to determine differences between recovery and delivery. Results: It was observed that the delivery of the targeting analyte was different from its recovery. Both delivery and recovery of puerarin were independent of the concentration of the drug. Probe delivery increased from 62.18 to 67.98 % (p < 0.01), recovery from 59.19 to 78.44 % (p < 0.01), as stirring speed was increased from 0 to 800 rpm. The difference between them directly correlated with stirring speed in the range 2.99 to 10.46 %. Besides additives, length of membrane also had a strong influence on delivery and recovery. Probe delivery in saline containing 10 % each of ethanol and propylene glycol declined from 62.18 to 42.12 % (p < 0.01), while recovery increased slightly but insignificantly (p < 0.01). Both delivery and recovery declined when the length of membrane was reduced. Conclusion: The in vitro experiments indicate that it would be incorrect to equate delivery with recovery of puerarin in in vivo microdialysis experiments

Investigation of Factors Affecting Microdialysis Probe Delivery and Recovery of Puerarin In Vitro

Purpose: To investigate in vitro the factors affecting microdialysis probe delivery and recovery of puerarin . Methods: The recovery and delivery of puerarin were tested for extraction efficiency and retro-dialysis methods. Factors such as drug concentration, stirring speed, additives and length of membrane were studied to determine differences between recovery and delivery. Results: It was observed that the delivery of the targeting analyte was different from its recovery. Both delivery and recovery of puerarin were independent of the concentration of the drug. Probe delivery increased from 62.18 to 67.98 % (p < 0.01), recovery from 59.19 to 78.44 % (p < 0.01), and stirring speed increased from 0 to 800 rpm. The difference between them directly correlated with stirring speed in the range 2.99 to 10.46 %. Besides, additives, length of membrane also had a strong influence on delivery and recovery. Probe delivery in saline containing 10 % each of ethanol and propylene glycol declined from 62.18 to 42.12 % (p < 0.01), while recovery increased insignificantlly (p < 0.01). Both delivery and recovery declined while the length of membrane was cut down. Conclusion: The in vitro experiments indicate that it was not correct to equate delivery with recovery of puerarin in in vivo microdialysis experiments

Studies of Mucosal Irritation and Cellular Uptake Mechanisms of Xingnaojing Nanoemulsion

Xingnaojing (XNJ) injection was used to treat pneumonia and stroke in clinic in China, but with poor patient compliance. Xingnaojing nanoemulsion for intranasal delivery was developed to improve it. This article tried to evaluate the mucosal irritation of Xingnaojing nanoemulsion and investigate cellular uptake mechanism of its encapsulated lipophilic drugs. The toad palate model and rat nasal mucosa model were used to study the nasal ciliotoxicity and nasal mucosal irritation of nanoemulsion to evaluate its safety intranasally. The cellular uptake mechanism was studied by Calu-3 cell model. Coumarin 6 was encapsulated in nanoemulsion and the endocytic pathways were studied by cellular uptake experiments after being treated with different inhibitors. In toad palate model, the cilia movement of Xingnaojing nanoemulsion group last for 467.40 ± 39.02 min, which was obviously longer than deoxycholate group (90.60 ± 15.40 min). Studies on rats showed that the damage caused by nanemulsion is capable of being recovered. Nanoemulsion uptake was reduced obviously when cells were treated with wortmannin, and it also decreased about 13% when the temperature reduced from 37ºC to 4ºC. Mucosal irritation caused by nanoemulsion is low and the damage is recoverable. The cellular uptake of Xingnaojing nanoemulsion is energy-dependent, and macropinocytosis was the most important pathway for cellular uptake