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OBJECTIVE:To establish a method for the simultaneous determination of 5 unsaturated fatty acids in Perilla oil cap-sule. METHODS:With the reference material of α-linolenic acid methyl ester,GC was used to determine and calculate the relative correction factors of α-linolenic acid methyl ester with methyl palmitate,methyl stearate,methyl oleate and linoleic acid methyl es-ter,and the correction factors were used to calculate the contents of 5 unsaturated fatty acids;the column was Agilent Innowax cap-illary column,the detector was FID,the inlet temperature was 230 ℃,the detector temperature was 250 ℃,the gas flow rate was 20 ml/min(nitrogen),40 ml/min(hydrogen)and 350 ml/min(air),split ratio was 30 to 1,the column temperature was 190 ℃, and injection volume was 1 μl. RESULTS:The linear range was 0.018-0.792 μg(r=0.9994)for methyl palmitate,0.0016-0.0176μg(r=0.9993)for methyl stearate,0.0056-0.2464 μg(r=0.9999)for methyl oleate,0.003-0.132 μg(r=0.9990)for linoleic acid methyl ester and 0.018-0.792 μg(r=0.9998) for α-linolenic acid methyl ester;RSDs of precision,stability and reproducibility tests were lower than 5%;recoveries were 98.990%-101.70%(RSD=0.720%,n=6) for methyl palmitate,99.599%-100.699%(RSD=0.368%,n=6) for methyl stearate,98.996%-101.680%(RSD=1.240%,n=6) for methyl oleate,99.813%-100.963%(RSD=0.434%,n=6)for linoleic acid methyl ester and 97.185%-99.602%(RSD=0.874%,n=6)for α-linolenic acid methyl es-ter. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determina-tion of methyl palmitate,methyl stearate,methyl oleate,linoleic acid methyl ester,α-linolenic acid methyl ester in Perilla oil cap-sule.
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This study aimed at investigating the antiviral constituents from the active fractions of Tong-An (TA) injection.In this study,the active constituents of TA injection were screened by LPS-induced PGE2 production mode to detect the contents of PGE2.The chemical constituents were isolated by HP-20 macroporous resin,silica gel column chromatography,ODS column chromatography,Sephadex LH-20 column chromatography and preparative and semi-preparative HPLC.The structures were identified by spectral data and physicochemical property.As a result,the 95％ ethanol eluate of TA injection on the macroporous adsorption resin column was proved to be the active fraction of TA injection.Seventeen compounds were isolated from TA injection and identified as syringaresinol (1),N-Trans-Feruloyltyramine (2),chelerythrine (3),sinomenine (4),coptisine (5),sanguinarine (6),chelidoniny (7),magnoflorine (8),allocryptopine (9),protopine (10),farrerol (11),dihydrosanguinarine (12),heptadec-(9Z)-enoic acid (13),chlorogenic acid (14),cryptochlorogenin acid (15),3,5-di-O-caffeoylquinic acid (16) and 4,5-di-O-caffeoylquinic acid (17).PGE2 inhibitory activities of these compounds were determined,among which six compounds presented inhibitory activities against PGE2.It was concluded that all the isolated compounds from TA injection were firstly reported with the favorable inhibitory activities of compounds 2,5,9,10,11,12 against PGE2.
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This paper was aimed to study the human plasma protein binding rate of diterpene ginkgolides meglumine injection.The equilibrium dialysis was used to determine the human plasma protein binding rate of ginkgolide A (GA),ginkgolide B (GB) and ginkgolide K (GK) in diterpene ginkgolides meglumine injection.The LC-MS/MS method was used for the content determination of ginkgolides.And then,the plasma protein binding rate was calculated.The results showed that there was no interference from other ingredients for the determination of ginkgolides.The calibration curve of the analytes was in good linearity in certain range of contents.The precision and stability of the analytes met the methodology requirements.After 8 h incubation,the human plasma protein binding rate of GA,GB and GK achieved balance.The human plasma protein binding rate of GA (0.34,1.70 and 8.51μg·mL-1) was 84.03%-88.11%; the human plasma protein binding rate of GB (0.62,3.09 and 15.5μg·mL-1) was 41.21%-53.56%; the human plasma protein binding rate of GK (0.04,0.20 and 1.01μg·mL-1) was 45.24%-59.59%.It was concluded that the method was simple,rapid and sensitive,which met the analysis requirement for biological samples.GA had a high plasma protein binding rate; GB and GK had medium plasma protein binding rate.
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This study was aimed to establish a simultaneous determination method of chlorogenic acid,liquiritin,rosmarinic acid,arctiin,glycyrrhizic acid,schisandrin,menthone in Guo-Ming-Xing Bi-Yan(GMXBY) granules by HPLC under multiple UV wavelengths.Waters Symmetry C18 column (4.6 mm× 250 mm,5μm) was used as the chromatographic column.Acetonitrile-0.1% phosphoric acid water solution was used as the mobile phase with gradient elution.The detection wavelength of chlorogenic acid and rosmarinic acid was 327 nm; that of liquiritin and arctiin was 280 nm; that of glycyrrhizic acid,schisandrin and menthone was 250 nm.The column temperature was 25℃.The injection volume was 10μL.Chlorogenic acid showed a good linear relationship in the range of 1.19-59.50μg?mL-1 (r = 0.999 8).The average recovery rate was 100.95%.Liquiritin showed a good linear relationship in the range of 1.51-150.70μg?mL-1 (r = 0.999 1).The average recovery rate was 100.38%.Rosmarinic acid showed a good linear relationship in the range of 3.40-68.08 μg?mL-1 (r = 0.999 9).The average recovery rate was 101.02%.Arctiin showed a good linear relationship in the range of 56.15-1 123.00μg?mL-1 (r =0.999 9).The average recovery rate was 100.39%.Glycyrrhizic acid showed a good linear relationship in the range of 21.54-430.80μg?mL-1 (r = 0.999 8).The average recovery rate was 97.09%.Schisandrin showed a good linear relationship in the range of 2.57-51.34μg?mL-1 (r = 0.999 9).The average recovery rate was 99.19%.Menthone showed a good linear relationship in the range of 0.50-10.00μg?mL-1 (r = 0.999 9).The average recovery rate was 100.35%.This established method was simple and reliable with good reproducibility,which can be used as the determination method of active components in GMXBY granules.
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This study was aimed to investigate the induction effect ofRe-Du-Ning (RDN) Injection on rat liver microsome CYP450 enzymes. SD rats were randomly divided into the solvent control group, positive control group as well as the low, middle and high dose group of RDN (1, 2, 4 mL·kg-1·d-1). After drugs were administrated continuously for 7 days, the rats were sacrificed. The liver was weighed and prepared to microsomes. Meanwhile, the liver coefficients of rats were calculated. And the protein content was detected by BCA method. Finally, activities of five important subtypes of CYP450 enzymes such as CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A1/2 were measured by the“cocktail” method. The results showed that the levels of liver coefficients, microsome yield rate and activities of CYP450 subtypes increased significantly in the positive control group compared with the solvent control group (P < 0.01). There was no significant difference on the levels of liver coefficients, microsome yield and protein content between the low and middle dose group of RDN. However, there was significant difference on the levels of liver coefficients and microsome yield in the high dose group (P < 0.05). In terms of the influence on enzyme activity, RDN Injection can significantly induce the activities of CYP1A2 with dose dependence. It can induce the activities of CYP2C9 and CYP2C19 at the middle and high dose. However, there was no obvious influence on the activities of CYP3A1/2 and CYP2D6. It was concluded that the positive control group can obviously induce activities of CYP450, which can be used in the evaluation of induction experiments. RDN Injection had induction effect on CYP1A2, CYP2C9 and CYP2C19. But it had no influence on the activities of CYP3A1/2 and CYP2D6.
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This study was aimed to establish a separation method for neochlorogenic acid reference substances from Lonicera japonica. Refined neochlorogenic acid inL. japonica water extract was separated and concentrated by HPD200A macroporous resin, which was isolated and purified by medium-low-pressure preparative chromatography and determined by HPLC. The structure was identified by various spectroscopic data including ESI-MS,1H-NMR and13C-NMR. The results showed that the optimal purification technology conditions were as follows: washed with 5BV of water, collected elution, concentration, drying; neochlorogenic acid crude products were eluted with acetonitrile-0.5% formic acid solution (10:90) with the flow rate of 20 mL·min-1; and the detection wavelength was 326 nm. The contents of the prepared neochlorogenic acid reached to 98.86% and the yield was 89.1%. It was concluded that the method was effective for the preparation of neochlorogenic acid with high purity. It can be used to prepare the reference substances for quantitative analysis and content determination of Chinese materia medica.
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Objective:To develop a method for the quantitative determination of ambrisentan. Methods: 1 H NMR spectra were obtalned with a Bruker AscendTM 400 superconducting NMR spectrometer. For each sample, DMSO-D6 was used as the solvent, the pulse width was 10. 0 μs, the delay time was 5 s and the scanning time was 16. Results: The proton peaks of ambrisentan at δ6. 16 ppm and maleic acid atδ6. 28 ppm were used as the quantitative peaks. The linear regression equation of peak area and quality ratio was Y=0. 140 7X+0. 034 8 with the correlation coefficient of 0. 999 4. RSD was 0. 2%(n=6)in the repeated experiments. The absolute content of ambrisentan reference substance was 99. 9%. Conclusion: The results showed that 1 H NMR can be used in the quantitative determination of ambrisentan without reference substance. The method is reliable, rapid, accurate and simple.
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Paeoniflorin and its derivatives are main active compounds inGui-Zhi Fu-Ling Capsule (GZFLC). In this study, molecular imprinted polymer (MIP) was prepared by sol-gel process to obtain paeoniflorin and its derivatives in GZFLC. The static adsorption capacity of MIP was measured by scatchard equation. The results showed that the maximum apparent absorbing capacity of MIP was 52.28 mg·g-1. One-step separation of paeoniflorin from 4 g methanol samples of GZFLC was 197 mg with the purity of 89.3%. It was concluded that paeoniflorin MIP can be used to separate phaoniforin and its analogues from GZFLC.
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To develop a method for the determination of 18 elements such as Pb, Cu, As, Hg, Mn, Ni, & Tl in Panax notoginseng,Bulbus fritillariae thunbergii,Coix seed, Resina draconis, and to control the contents of heavy metal elements in Sanjie Zhentong Capsule, the samples were digested by microwaves and then analyzed by appropriate determination parameters through ICP-MS, with the internal standard method to improve the matrix effect and interference. The correlation coefficientR2≥ 0.999 2. The lowest limits of quantification were from 0.002 8 to 0.54 μg·L-1. The experiments had better repeatability, while the recovery values ranged from 73.01% to 109.13%. The method is simple, accurate and high sensitive, and it can be used for the determination of rapid monitoring the multi-elements inPanax notoginseng, Bulbus fritillariae thunbergii, Coix seed,and Resina draconis.
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In this article, an HPLC method for the contents determination of amino acids in Qihong Maitong injection was reported. In detailed, OPA-Fmoc pre-column derivatization was adopted, and related HPLC methods to determine the contents of amino acids was established. Linear relationship was well constructed for 17 amino acids through the method mentioned above. Briefly speaking, the optimized method was accurate and reproducible, and suitable for the determination of amino acids in Qihong Maitong injection and corresponding quality control.
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<p><b>OBJECTIVE</b>To observe the anaphylaxis and hemocytolysis of 6 kinds of tween-80 injection from different source.</p><p><b>METHOD</b>The Hartley albinism guinea pig was used to carry on the active systematic anaphylaxis (ASA) and the passive cutaneous anaphylaxis (PCA). The in vitro hemolytic experiment and the hemocytolysis was observed by means of spectrophotometry on the domestic rabbit.</p><p><b>RESULT</b>The result of ASA and PLA of 6 kinds of tween-80 injection from different source assumed the negative. In observation time, the temolysis rate of 3 kinds of tween-80 injection are more than 5%, while the others are also more than 5% only in the highly concentrated test tube.</p><p><b>CONCLUSION</b>Six kinds of tween-80 injection from different source have not caused the immune-mediated anaphylaxis, but it may have hematolysis tendency on intravenous injection. The hemocytolysis of tween-80 may not be entirely caused by the impurities. It is worthy of further study that the physical and chemical properties of the product itself and the undeserved concentration is doubtful whether there is also some internal relations with the generation of hemolytic.</p>
Subject(s)
Animals , Female , Male , Rabbits , Anaphylaxis , Chemistry, Pharmaceutical , Guinea Pigs , Hemolysis , Injections , Passive Cutaneous Anaphylaxis , Polysorbates , ChemistryABSTRACT
AIM:To study the chemical constituents of the roots of Astragalus hoantchy Franch.. METHOD:Isolation and elucidation of the chemical constituents,were conducted by chromatography and spectral evidences. RESULTS and CONCLUSION:Six steroids and four anthraquinones were isolated from the roots of A. hoantchy. Their structures were identified to be stigmastane-3,6-dione (1),5α,8α-epidioxy- (22E,24R)-ergosta-6,22-dien-3β-ol (2),stigmastane-3β,6α-diol (3),daucosterol (4),β-sitosterol (5),stigmasterol (6),chrysophanol (7),emodin (8),physion (9) and aloe-emodin (10) on the basis of spectral data and physical constants. Among them,compounds 1,2,3,7,8,9,10 were isolated from the genus Astragalus for the first time.