Journal of Chinese Pharmaceutical Sciences

Chemical profiling of Scutellaria barbata by ultra high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry

Ru Li, Wei Song, Xue Qiao, Jia Liu, Hong Liang, Min Ye

2015, 24(10): 635-646. DOI: 10.5246/jcps.2015.10.081

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An ultrahigh performanceliquid chromatographycoupled with hybrid quadrupole-orbitrap mass spectrometry (UHPLC-quadrupole-orbitrap MS) method was developed to systematically analyze chemical constituents of Scutellaria barbata D. Don. The 70% methanol extract was separated on an Agilent Eclipse Plus C₁₈ column (1.8 μm, 150 mm×2.1 mm), and eluted with a gradient of methanol–acetonitrile–water containing 0.1% formic acid at a flow rate of 0.25 mL/min. Constituents were identified by HRMS in the negative ion mode using both full scan and two-stage threshold-triggered mass modes. A total of 56 compounds, including 20 free flavonoids, 20 flavonoid O-glycosides, 2 flavonoid C-glycosides, 4 phenylethanoid glycosides, and 10 phenolic compoundswere identified by analyzing their high resolution mass spectral data.Among them, 8 flavonoids and 2 phenylethanoid glycosides were unambiguously identified by comparing with reference standards. This study demonstrates that hybrid quadrupole-orbitrap mass spectrometry is a powerful tool in structural identification of unknown compounds in complicated herbal extracts.

Anti-inflammatory effect of ethyl acetate extract of Sedum aizoon L. in LPS-stimulated RAW 264.7 macrophages and its HPLC fingerprint

Zhucan Lin, Long Zhang, Ruizhuo Zhang, Anyu Huang, Suhua Guo

2015, 24(10): 647-653. DOI: 10.5246/jcps.2015.10.082

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Sedum aizoon L. (SA) has been widely used for treatment of various hemorrhages, insomnia, pain and trauma; however, its anti-inflammatory activity is unknown. In this study, wefirstly investigated the anti-inflammatory activity of SA extracts by petroleum ether (PE), ethyl acetate (EtOAc), and H₂O in LPS-stimulated RAW 264.7 cells. Results showed that the EtOAc extract rich in phenolics and flavonoids significantly inhibited LPS-induced NO, TNF-α, and IL-6 production in RAW 264.7 cells (P<0.01), suggesting that this extract possessed potent anti-inflammatoryactivity. The phytochemical profile of the effective extract was subsequently analyzed by HPLC fingerprint with 11 standards. The results indicated that gallic acid, protocatechuic acid, p-hydroxybenzoic acid, ethyl gallate, iriflophene, 5,7-dihydroxy chromone, quercitrin, quercetin, luteolin, kaempferol, and isorhamnetin were present in this extract, which might contribute to its anti-inflammatory activity. Thus, the EtOAc extract of SA showed anti-inflammatory activity and could be used as a potential natural anti-inflammatory agent.

Chemical analysis of the Hedysarum multijugum root by HPLC fingerprinting

Yi Liu, Wei Wang, Yuying Zhao, Hubiao Chen, Hong Liang, Qingying Zhang

2015, 24(10): 654-659. DOI: 10.5246/jcps.2015.10.083

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The root of Hedysarum multijugum(RHM) is recorded as a folk herbal medicine in China and is sometimes used asa substitute for Hedysari Radix, which is a famous traditional Chinese medicine derived from the roots of Hedysarum polybotrys. In the present study, a sensible, reliable, and reproducible HPLC-DAD fingerprint analysis method for RHM was developed and then subsequently applied to analyze RHM samples from different origins. The chemical constituents of the RHM sampleswere generally consistent, although it wasslightly affected by the local environment of the plant. In addition, the chemical constituency of RHM was shown to be significantly different from that of Hedysari Radix, suggesting that RHM is not suitable as a substitute for Hedysari Radix, at least from the chemical point of view.

The targeting effect of H7K(R2)2-modified pH-sensitive liposomes on U87-MG cells

Yang Zhao, Wei Ren, Ting Zhong, Chao Wang, Weiqiang Zhang, Dan Huang, Shuang Zhang, Yang Guo, Xin Yao, Xuan Zhang, Qiang Zhang

2015, 24(10): 660-665. DOI: 10.5246/jcps.2015.10.084

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The present study aimed to investigate the targeting effect of H₇K(R₂)₂-modified pH-sensitive liposomes on U87-MG cells. Using coumarin-6 as a fluorescence probe, we prepared H₇K(R₂)₂-modified pH-sensitive liposomes (designated as coumarin-6-PSL-H₇K(R₂)₂). The flow cytometry assay was used to evaluate the effect of H₇K(R₂)₂proportions on the cellular uptake and endocytosis pathways of coumarin-6-PSL-H₇K(R₂)₂ on U87-MG cells_.The circular dichroism (CD) spectroscopy assay was used to investigate the secondary structures of H₇K(R₂)₂ peptide at pH 7.4 and pH 6.8, respectively. Our results indicated that the 2.5% proportion of H₇K(R₂)₂ in the coumarin-6-PSL-H₇K(R₂)₂ was superior to those of 1% and 3.5% of H₇K(R₂)₂. The uptake of coumarin-6-PSL-H₇K(R₂)₂ on U87-MG cells was not inhibited by filipin, M-β-CD or chlorpromazine. The secondary structure of H₇K(R₂)₂at pH 6.8 was mostly presented as β-turn. In conclusion, we suggested that the appropriate proportion of H₇K(R₂)₂ in the H₇K(R₂)₂-modified pH-sensitive liposomes could be set at 2.5%. The cellular uptake pathway for H₇K(R₂)₂-modified pH-sensitive liposomes was via the cell penetrating capacity of H₇K(R₂)₂ which responded to acidic condition. The secondary structure of H₇K(R₂)₂at pH 6.8, which was presented as the shape of hairpin, might be mainly responsible for its targeting and cell penetrating effect.

Folate and cell penetrating peptide modification effects on penetration behavior of nanostructured lipid carriers using multicellular tumor spheroids

Hongmeng Zhuang, Tingting Meng, Wei Gao, Lan Yuan, Xianrong Qi

2015, 24(10): 666-672. DOI: 10.5246/jcps.2015.10.085

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Surface modification may have important influences on the penetration behavior of nanoscale drug delivery system. In the present study, we mainly focused on whether cell targeting or cell penetration could affect penetration abilities of nanostructuredlipid carriers (NLC). Real-time penetration of folate- or cell penetrating peptide (CPP)-modified NLC was evaluated using a multicellular tumor spheroid (MTS) established by stacking culture method as an in vitro testing platform. The results suggested that CPP modification had a better penetration behavior both on penetration depth and intensity compared with folate-modified NLC at the early stage of penetration process.

Quality assessment and dissolution profile comparison studies on 250-mg mefenamic acid tablets available in local market of Karachi

Farya Zafar, Huma Ali, Shabana Naz Shah, Safila Naveed, Shehla Siddiqui

2015, 24(10): 673-677. DOI: 10.5246/jcps.2015.10.086

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In the present study, we aimed todetermine the quality of different brands (Test₁–Test₆) of mefenamic acid, which are commercially available in local market of Karachi, Pakistan. Various quality evaluation tests, including weight variation, hardness, thickness, friability, disintegration, assay and drug release, were carried out. Results were found to be in acceptable limits. Moreover, release profiles were compared using different dissolution media, such as phosphate buffer pH 6.8, 7.4 and biorelevant media (FaSSGF, FaSSIF, FeSSIF and SCoF). Release profiles at pH 6.8 and 7.4 were evaluated by one-way ANOVA, model-independent and model-dependent methods. Results of ANOVA showed that no significant difference was found among tester and reference brands (Test₁–Test₆). Similarly, all the brands were found to be best fitted with Weibull model.

Clinical efficacy and safety of intravenous itraconazole in patients with invasive fungal infections in emergency intensive care unit

Xiao Luo, Hong Shao

2015, 24(10): 678-682. DOI: 10.5246/jcps.2015.10.087

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This study aimed to analyze the clinical efficacy and safety of itraconazole.We investigated 68 patients with invasive fungal infections (IFI) in emergency intensive care unit (EICU). A retrospective analysis was performed in patients with IFI who were treated in the authors’ institution, a grade III first class hospital in Beijing, China, between Feb. 2013 and Feb. 2015. The age of patients ranged from 35 to 90 years old with the mean age of (75.1±11.1) years old. The study population comprised 36 male and 32 female patients. Total response rate was 60.3%. The response rates in definitive diagnosis, clinical diagnosis and presumed diagnosis were 33.3%, 59.5% and 65.2%, respectively (P<0.05). The empirical treatment should be provided for patients with presumed diagnosis as earlier as possible. Eleven (16.2%) cases had adverse drug event (ADE) during treatment. The main signs were hepatic functional impairment and hypokalemia of ADE. The clinical efficacy and safety of intravenous itraconazole were precisely assessed. To reduce the adverse drug reaction (ADR), hepatic and renal function and other biochemical criterion should be closely monitored.

Investigating on the effects of national essential medicine policy on primary care facilities in Hebei province: a case study

Zhao Yang, Yile Zhao, Jing Chen, Jiaxuan Yu, Jianwen Lan, Bin Jiang

2015, 24(10): 683-689. DOI: 10.5246/jcps.2015.10.088

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National essential medicine policy (NEMP) is an important part of new health care reform and core content of national drug policy. We chose Hebei province as a case to study, utilized standard methods from WHO/HAI and built interrupted time series (ITS) model to qualitatively and quantitatively evaluate the effects of NEMP in Hebei province from the utilization of essential medicines. Shortly after implementing EMP, the purchasing and utilization rate of essential medicines significantly increased, but no further continuous effects. In order to perfect the essential medicine policy, training of rational drug utilization should be strengthened, hierarchical essential medicine list and dynamic monitoring on the effect of NEMP are necessary.

Determination of chlorpromazine in porcine muscle using high performance liquid chromatography-tandem mass spectrometry

Lu Zhang, Wen Huang, Xinhui Jiang

2015, 24(10): 690-694. DOI: 10.5246/jcps.2015.10.089

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A rapid and accurate high performance liquid chromatography tandem mass spectrometry (HPLS-MS) method was established for quantification of chlorpromazine in pork. The porcine samples were pretreated with acetonitrile to precipitate proteins and followed by extraction with tert-butyl methyl ether (TBME). The separation was performed on a 5 µm Agilent XDB-C₁₈ column with gradient elution. The mobile phase A was 0.01 mol/Lammonium formate in water and mobile phase B was acetonitrile. The flow rate was at 0.8 mL/min. Quantification was performed on a triple-quadrupole tandem mass spectrometer using the multiple selected reaction monitoring (MRM) mode. Transition of m/z +319.1 to 58.1 was used for the quantification of chlorpromazine. The method was validated at the concentration range of 0.4040 µg/kgto 8.080 µg/kg for chlorpromazine with correlation coefficient of 0.9999. The spiked recoveries were more than 80.0% and the limit of detection (LOD) was 0.052 µg/kg.The developed method, which offers advantages of convenience, rapid, specificity and higher sensitivity, can be used for determination of chlorpromazine hydrochloride in porcine muscles.

Information from US FDA

http://www.fda.com

2015, 24(10): 695-698.

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