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Establishment of LC-MS/MS method for the determination of forsklin in rat plasma and its pharmacokinetics

Dianwei SONG ; Decai WANG ; Zhiyun MENG ; Ruolan GU ; Meihui SHI ; Zhuona WU ; Jingze WANG ; Guifang DOU

Journal of International Pharmaceutical Research.2012;(2):149-153. doi:10.3969/j.issn.1674-0440.2012.02.010

Objective To develop a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of forsklin in rat plasma.Methods After extraction with methyl tert-butyl ether,chromatographic separation was performed on a C18 column with the mobile phase consisting of water ( 0.1％ formic acid)-acetonitrile in a gradient elution mode.A tandem mass spectrometer equipped with electrospray ionization (ESI) source was used as detector in the positive ion mode.Quantification was performed using multiple reaction monitoring (MRM) with the precursor product combination ions of m/z 411→375.3 and 285→193 for forsklin and diazepam.Results Good linearity was obtained in the 0.5-1000 ng/ml range for the analyte and the analytical method was validated in terms of specificity,precision,accuracy,recovery,stability and matrix effect.These assays gave RSD values always lower than 14.4％ and RE values between -3.5 ％ and 3.8％.In addition,the specificity,extraction recovery,stability and matrix effect were satisfactory.Conclusion Due to its high sensitivity,specificity and simplicity,the method could be used for pharmacokinetic studies of forsklin.

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Determination of aconitine in dog tissue homogenates by HPLC-MS/MS and its application to in vitro metabolic stability study

Cuiping YANG ; Sha LIAO ; Tianhong ZHANG ; Jinglai LI ; Xiaoying WANG ; Jinxiu RUAN ; Zhenqing ZHANG

Journal of International Pharmaceutical Research.2012;(3):256-260. doi:10.3969/j.issn.1674-0440.2012.03.015

Objective To develop a HPLC-MS/MS method for the determination of aconitine and study thein vitro metabolic stability of aconitine in dog tissue homogenates.Methods The chromatographic separation was performed on a C18 column.The mobile phase consisted of acetonitrile and water with 0.2％ formic acid and 5 mmol/L ammonium acetate.A triple quadrupole tandem mass spectrometer equipped with an electrospray ionization interface source was used for the quantitative determination in the positive selective reaction monitor mode.Aconitine was incubated with dog tissue homogenates and samples were withdrawn at different time points and precipitated by acetonitrile with internal standards citalopram.Results Aconitine showed good linear relationship over the range from 5 to 500 ng/ml.The recoveries of aconitine were between 85.73％ and 92.12％ at three QC concentration levels.The intra- and inter-day precisions were 5.32％ - 8.95％ and 5.45％ - 8.86％,respectively.After incubation,about 20％ of aconitine were cleared in the liver and small intestine,and t1/2 were 460.6 and 521.3 min,respectively.But none was metabolized in the stomach and kidney.Conclusion These results demonstrated that aconitine was mainly metabolized in the liver and small intestine at a slow rate.

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Effect and possible mechanism of Albizia chinensis extract on gastric ulcer in rats

Yu ZHANG ; wei Xiao ZHOU ; ming Rui XU ; shan Shi YU ; gang Shuang MA ; jun Jian ZHANG

Journal of International Pharmaceutical Research.2017;44(8):783-789. doi:10.13220/j.cnki.jipr.2017.08.007

Objective To investigate the gastric protective effect of Albizia chinensis extract on acute gastric ulcer model in rats and its mechanism related to H+,K+-ATPase in gastric parietal cells. Methods Rats were injected intraperitoneally with indo?metacin or kept under water immersion-restraint stress(WRS)to establish acute gastric ulcer model. Test samples were orally given 30 min before induction of gastric ulcer,and the gastric ulcer index and inhibitory rate were used to evaluate the effect of A. chinensis extract. Gastric mucosa pathological tissues in rats were observed by HE staining,the enzymatic activity of proton pump on isolated gastric parietal cells was measured,and the translocation of gastric proton pump was observed by immunofluorescence cell staining. Results The A. chinensis extract inhibited the gastric ulcer with the inhibition rate of 70.0%and 75.4%at 300 mg/kg in the WRS and indomethacin-induced rat model,respectively. The extract could also inhibit the enzymatic activity of gastric proton pump with the in?hibition rate of 21.5%,29.4%,44.1%,51.9%,57.7%and 62.9%(P<0.01)at 15,30,60,120 and 480 mg/L,respectively. The half-effective dosage(ID50)of the extract was determined to be 130 mg/L. The extract showed no significant effect on the translocation of gastric proton pump. Conclusion The A. chinensis extract(300 mg/kg,ig)could significantly prevent the formation of gastric ul?cer in rats via inhibiting the enzymatic activity of gastric proton pump.

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Biodegradable microneedles applied in the transdermal drug delivery:research advances

feng Qian HUANG ; qing Hua LIN ; xin Bin PENG ; zhou Fang SHUAI

Journal of International Pharmaceutical Research.2017;44(8):778-782. doi:10.13220/j.cnki.jipr.2017.08.006

Microneedle is a painless and minimally invasive transdermal drug delivery. It can enhance the permeability of skin for macromolecular drugs by puncturing and creating a micron-level drug delivery channel in the skin. The biodegradable microneedles aim to solve the problem that nondegradable-material microneedles may break under skin and cause adverse reaction. This article pro?vides an overview of the characteristics of biodegradable microneedles,the biodegradable materials used,its recent research progress, the development status and the tendency.

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MicroRNA for preventing and treating diabetes mellitus complications:research advances

hao Wen YANG ; ping Yan LEI

Journal of International Pharmaceutical Research.2017;44(8):772-777. doi:10.13220/j.cnki.jipr.2017.08.005

Diabetes mellitus is a chronic disease critical to human health. Its complications can lead to some organs and tis?sues damage and are the major cause of death in diabetic patients. As a new type of diagnosis and treatment ,microRNA(miRNA)play an important role in the pathogenesis of diabetes mellitus and its complications. MiRNAs associated with diabetes are becoming an im?portant therapeutic target for diabetic complications. This review summarizes the effects of miRNAs on the pathogenesis of diabetic car?diomyopathy and diabetic nephropathy,and discusses its role in the diagnosis and treatment of diabetes mellitus and its complications.

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Comparative analysis of drug intervention effect on patients with impaired glucose regulation in China

Feng GU ; wen Yao LIU ; liang Xiao ZHANG ; jie Jun SHAN

Journal of International Pharmaceutical Research.2017;44(8):765-771. doi:10.13220/j.cnki.jipr.2017.08.004

Impaired glucose regulation(IGR,also named prediabetes)is an abnormal condition and important period for dia?betes mellitus. The period includes impaired fasting glucose or/and impaired glucose tolerance. IGR should be screened promptly and be interfered by life style and drugs to recover normal condition,or to delay and control the development of diabetes mellitus for de?creasing the highrisk of diabetes mellitus and cardiovascular disease. The paper reviews the literature about antiglycemic drugs and tra?ditional Chinese medicine interventions for IGR in China in the recent ten years,and summarizes some characters of these drugs to provide some help for prediabetes.

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Simultaneous determination of nine components in Kanglixin Jiaonang by HPLC wavelength switching combined gradient elution method

yang Si LI ; xin Qing KONG

Journal of International Pharmaceutical Research.2017;44(8):817-822. doi:10.13220/j.cnki.jipr.2017.08.013

Objective To develop an HPLC wavelength switching combined gradient elution method for simultaneous determi?nation of nine components in Kanglixin Jiaonang(KLXJN). Methods The analysis was performed on a Kromasil C18(4.6 mm×250 mm, 5μm)with gradient elution by using the mobile phase of acetonitrile-methanol(1:2)(A)-0.1%phosphoric acid solution(B). The col?umn temperature was maintained at 30℃and the flow rate was 0.9 ml/min. The detection wavelengths were set at 225 nm for costuno?lide(1)and dehydrocostus lactone(2),254 nm for aloe-emodin(3),rhein(4),emodin(5)and physcion(6),and 425 nm for bisde?methoxycurcumin(7),demethoxycurcumin(8)and curcumin(9). Results The calibration curves were linear within the range(μg/ml)of 6.610-132.2(r=0.9999)for 1,7.890-157.8(r=0.9996)for 2,14.07-281.4(r=0.9992)for 3,3.450-69.00(r=0.9997)for 4, 2.670-53.40(r=0.9998)for 5,3.760-75.20(r=0.9999)for 6,5.880-117.6(r=0.9996)for 7,8.490-169.8(r=0.9993)for 8,and 13.91-278.2(r=0.9991)for 9,respectively. The recoveries for 1,2,3,4,5,6,7,8 and 9 were 98.28%(RSD=1.09%),97.76%(RSD=0.80%),99.08%(RSD=1.72%),97.19%(RSD=1.00%),98.45%(RSD=1.24%),96.96%(RSD=1.21%),98.51%(RSD=1.55%), 97.52%(RSD=0.83%),and 100.04%(RSD=0.93%),respectively. Conclusion The established method is accurate,rapid and can be used for the quality control of KLXJN.

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Simutaneous determination of ursolic acid and oleanolic acid in Ziziphora clinopodioides Lam. by in situ pretreatment-thin layer chromatography scanning

huan Wen DING ; yin Tian YANG ; jia Xue ZHANG ; yan Hai XU ; ying Xiao ZHOU

Journal of International Pharmaceutical Research.2017;44(8):812-816. doi:10.13220/j.cnki.jipr.2017.08.012

Objective To establish a method for the simultaneous determination of ursolic acid(UA)and oleanolic acid(OA) in Ziziphora clinopodioides Lam.,and the quantitative determination the UA and OA contents in the different Z. clinopodioides plant samples collected with various parts of the plant at different times,from different regions of Xinjiang,China. Methods Dual wave?length scanning method was used for the quantification of UA and OA spots on a silica gel G plate in the TLC analysis. The samples loaded on the silica gel G plate were in situ treated with the 1%iodine solution in dichloromethane,and then the plate was developed using cyclohexane,cyclohexane-chloroform-ethyl acetate-formic acid(20:5:8:0.1)as the developing solvent. In the dual wavelength scanning,the measurement wavelength was 530 nm and the reference wavelength was 700 nm. Results The UA and OA spots in samples were well separated on the TLC plate and could be simultaneously quantified by the present method. The average contents of UA and OA in Z. clinopodioides plant samples from 18 different areas were(1.84 ± 0.41)and(2.82 ± 0.89)mg/g,respectively. The contents of UA and OA in the plant increased from late spring to early summer and then decreased thereafter. As to the different parts of the plant,the contents of UA and OA were highest in leaves and lowest in stems. Conclusion The method is simple,fast and accu?rate. The present results provided basic data for further evaluation of the quality of Z. clinopodioides resources.

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HPLC determination of the content and dissolution of isophenylcyclopentylamine hydrochloride capsules

fang Yue ZHOU ; zhou Yun FAN ; ke Xiao WANG ; you Yan LI ; ping Xiao CHEN ; sheng Chun GAO

Journal of International Pharmaceutical Research.2017;44(8):807-811. doi:10.13220/j.cnki.jipr.2017.08.011

Objective To establish a method for the determination of the content and dissolution of isophenylcyclopentyl?amine hydrochloride capsules. Methods The HPLC analysis was performed on a Diamonsil C18 column(150 mm×4.6 mm,5μm). A mixture of 0.02 mol/L KH2PO4 solution containing 0.1%triethylamine with pH adjusted to 3.0 by phosphoric acid-methanol-acetonitrile (30:35:35)was used as the mobile phase with the flow rate at 1.0 ml/min. The detection wavelength was 224 nm. Dissolution was de?termined by the basket method,using the 500 ml of 0.1 mol/L hydrochloric acid solution,pH 4.5 acetic acid buffer,water and pH 6.8 phosphoric acid buffer as dissolution media under the 50,75 and 100 r/min rotation speeds to select the dissolution condition. Re?sults This method had high specificity. The linear range for the quantitative determination was 20.74-155.58 μg/ml(r=1.0000), and the average recovery was 100.1%. The linear range for the determination of dissolution was 2.08-24.90μg/ml(r=0.9998),with the average recovery of 98.9%. The method of dissolution tests was established:0.1 mol/L hydrochloric acid solution was used as disso?lution medium and rotation speed was 50 r/min. The determined content and dissolution of three batches of capsules fulfilled the re?quirements. Conclusion The method is simple,accurate and reproducible for the determination of the content and dissolution of iso?phenylcyclopentylamine hydrochloride capsules.

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Response surface methodology applied to the optimization for supercritical CO2 extraction of the lipids from tempeh

ming Zi XIA ; Ying TIAN ; hui Xuan HE ; xing Jun DONG

Journal of International Pharmaceutical Research.2017;44(8):800-806. doi:10.13220/j.cnki.jipr.2017.08.010

Objective To optimize the supercritical carbon dioxide(SC-CO2)extraction of lipids from tempeh(TE-C)and further improve the lipid classes ratio. Methods The experimental parameters of SC-CO2 extraction including extraction temperature, pressure,and moisture content of tempeh were optimized using a Box-Behnken design combined with response surface methodology (RSM),according to the weighted extraction ratio of TE-C and lipid classes after the experimental results of single factors. Detailed chemical compositions of TE-C obtained by optimum conditions of SC-CO2 extraction were analyzed by high performance liquid chroma?tography with an evaporative light-scattering detector(HPLC-ELSD)and high performance liquid chromatography-atmospheric pres?sure chemical ionization mass spectrometry(HPLC-APCI-MS). Results TE-C was composed of three lipid classes:fatty acids(Ⅰ), diacylglycerols(Ⅱ)and triacylglycerols(Ⅲ). The optimum SC-CO2 extraction conditions of TE-C were 50℃extraction temperature, 25 MPa pressure,1.99%moisture content of tempeh and 1.5 hour extraction time. Conclusion The optimum value of RSM for SC-CO2 extraction was(5.97±0.15)g/100 g.

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