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OBJECTIVE：To obs erve the protective effect of protoca techuic aldehyde（PAL）on neurovascular unit （NVU） homeostasis damage in rats after cerebral ischemia-reperfusion injury （CIRI）. METHODS ：SD rats were randomly divided into sham operation group ，model group ，PAL high-dose and low-dose groups （10，20 mg/kg），with 11 rats in each group. Administration groups were given relevant medicine intragastrically. Sham operation group and model group were given the same volume of normal saline intragastrically ，10 mL/kg once a day ，for 5 days. After last administration ，CIRI model was induced by suture method ；the ultrastructural changes of NVU were observed by transmission electron microscope. Western blot assay was used to detect the expression of NUV related proteins （MAP-2，GFAP，AQP-4）in cerebral tissue. Immunofluorescence staining was used to observe the positive expression of above proteins in cerebral cortex. RESULTS ：Compared with sham operation group ， blood-brain barrier （BBB）structure of model group was destroyed severely ，the vascular lumen became narrower ，lateral edema of endothelial cells was severe ，and the thickness of basement membrane varied ；the nuclei of neurons were pyknosis and there was a large area of edema in the surrounding tissues ；the structure of glial cells was seriously damaged ，the cell body was shrunk and organelles were lost ；protein expression （or positive expression ）of MAP- 2 in brain tissue （or cerebral cortex ）were significantly decreased （P＜0.05 or P＜0.01），while protein expression （or positive expression ） of GFAP and AQP- 4 were increased significantly（P＜0.01）. After PAL intervention ，the rats had less BBB damage ，and the morphology of vascular lumen and basement membrane were not completely destroyed ；the damage of neurons was alleviated ，the pyknosis of neurons was decreased ， the chromatin was homogeneous and the heterochromatin was decreased；the damage of glial cell structure was alleviated protein expression of GFAP and AQP- 4（except for low-dose group） in cerebral tissue and positive expression of MAP- 2 and GFAP protein in cerebral cortex were reversed @qq.com significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：PAL can protect the stability of NVU from damage in CIRI model rats； the mechanism may be related to up-regulating the expression of MAP- 2 protein in cerebral cortex and down-regulating the expression of GFAP and AQP- 4 protein in brain tissue.
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Objective: To optimize the water-precipitation in the pretreatment manufacture process of Salviae Miltiorrhizae (Danshen) and Ligustrazine Hydrochloride Injection (SMLHI) based on the quality by design (QbD) concept and the sequential design. Methods: A design space was obtained by the global-type sequential design methodology combined with the fishbone-diagram risk analysis and central composite experiment based on global type sequential design method. The preliminary optimized zone of critical process parameters was extended in axial direction on the basis of the statistical result gained from the first central composite design. Then an updated multivariate linear regression model was fitted and a design space was delineated and verified. Results: According to the design space, the recommended operation space of the water-precipitation was as follows: when the pH value of the material was between 3.1 and 3.4, 3.25-5.00 times of water adding into the solution was recommened, the standing time was 7-17 h and the standing temperature was 7 ℃. Conclusion: This study introduces the sequential design into the process optimization of traditional Chinese medicine pharmaceuticals. Compared with the experimental design methods such as uniform space grid design and orthogonal design with a large number of samples, while samples cover the same feasible zone in a continuous operation form, sequential design methodology can effectively reduce the workload of the optimization experiment, avoid the waste of manpower and resources as well as guarantee the prediction ability of the fitting model.
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Objective: To observe the protective effect of Salvia miltiorrhizae and Ligusticum chuanxiong effective constituents: danshensu, protocatechuic aldehyde, ligustrazine, and ferulic acid combination on primary cultured hippocampal neurons injured by oxygen glucose deprivation, and find out an optimized combination. Methods Primary cultured rats hippocampal neurons waschosen as research objects by adopting immunohistochemistry of the neuron-specific enolase IgG to authenticate, then the OGD model of the hippocampal neurons injured by oxygen glucose deprivation was established. The non-cytotoxic dose range of danshensu, protocatechuic aldehyde, ligustrazine, ferulic acid, and nimodipine was studied by MTT method. The compatibility of components was arranged by L9 (34) orthogonal design. Primary cultured rats hippocampal neurons was divided into 12 groups: control group, model group, Nimodipine positive control group, and orthogonal design 1-9 group. The activity of LDH was measured by colorimetry, the activity of SOD was tested by WST-1 and the levels of MDA were examined by TBA. The levels of TNF-α, IL-1β, and IL-6 in cell culture supernate were examined by ELISA, the apoptosis of hippocampal neurons was detected by Fluorochrome Hoechst33258 staining and the cell early apoptosis rate was detected with flow cytometry. The results of orthogonal test was analyzed by using range analysis method. Results The orthogonal compatibility of reagents played significant roles against the hypoxia damage of hippocampal neurons, improved the cellular morphology obviously, reduced the activity of LDH, increased the activity of SOD and decreased the content of MDA significantly, inhibited the release of TNF-α, and reduced the content of IL-1β and IL-6, reduced the apoptosis of cells apparently. The effect of active ingredients of S. miltiorrhiza and L. Chuanxiong on LDH activity was danshensu > ligustrazine > protocatechuic aldehyde > ferulic acid, and the best combination was danshensu (120 μg/mL), protocatechuic aldehyde (120 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (20 μg/mL). The effect on SOD activity was ferulic acid > ligustrazine > danshensu > protocatechuic aldehyde. The best combination was danshensu (120 μg/mL), protocatechuic aldehyde (120 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (40 μg/mL). The order of influence on MDA content was danshensu > protocatechuic aldehyde > ferulic acid > ligustrazine, and the best combination was danshensu (60 μg/mL), protocatechuic aldehyde (60 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (20 μg/mL). The effect on TNF-α content were: ligustrazine > protocatechuic aldehyde > danshensu > ferulic acid, and the best combination was danshensu (60 μg/mL), protocatechuic aldehyde (60 μg/mL), ligustrazine (40 μg/mL), and ferulic acid (10 μg/mL). The order of influence on the content of IL-1β was ligustrazine > ferulic acid > danshensu > protocatechuic aldehyde, and the best combination was danshensu (30 μg/mL), protocatechuic aldehyde (30 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (20 μg/mL). The effect on the content of IL-6 was protocatechuic aldehyde > ligustrazine > ferulic acid > danshensu, and the best combination was danshensu (120 μg/mL), protocatechuic aldehyde (120 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (10 μg/mL). The effect on the early apoptosis rate was ferulic acid > protocatechuic aldehyde > ligustrazine > danshensu, and the best combination was danshensu (60 μg/mL), protocatechuic aldehyde (30 μg/mL), ligustrazine (20 μg/mL), and ferulic acid (40 μg/mL). Conclusion The protective mechanism of the effective constituents of S. miltiorrhizae and L. chuanxiong were relied on reducing the oxidative damage, reducing inflammation damage, and antagonizing cell apoptosis. According to the experimental results, we need to change the prescription ratio and guide clinical medication for different clinical courses.
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Objective: To study the effect of 60Co-γ ray irradiation on the contents of danshensu,protocatechuic aldehyde,puerarin and salviamolic acid B in Tongmai granules by HPLC. Methods: An Agilent ZORBAX Eclipse XDB C18(2) column (250 mm × 4. 6 mm,5 μm) was adopted and the wavelength of UV detection was 280 nm at the flow rate of 1. 0 ml·min-1. The mobile phase consisted of 0. 1% phosphoric acid (B) and acetonitrile (A) with gradient elution, and the column temperature was 35℃. Tongmai granules were irradiated by 60Co-γ ray respectively at 0, 2, 5,10 and kGy,the contents of the active ingredients were compared before and after the irradiation. Results: The linear range of danshensu,protocatechuic aldehyde,puerarin and salviamolic acid B was 0. 098-4. 925 μg, 0. 028-1. 411 μg, 0. 378-18. 882 μg and 0. 218-10. 888 μg, respectively. The average recovery was 99. 8% , 97. 7% , 99. 9% and 99. 9% , respectively. When the radiation dose was not more than 2 kGy, the contents of the five components did not change significantly (P>0. 05). After 5 kGy radiation, the contents of protocatechuic aldehyde and salviamolic acid B were signifi-cantly different (P <0. 05). Conclusion: The dose of 60Co ray should be controlled not more than 2 kGy, and the sterilization method is safe and effective for Tongmai granules.
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Objective To investigate the chemical constituents from salvia polyphenolic acids. Methods Various column chromatographic techniques were used to separate and purify the chemical constituents and their structures were identified by spectral analysis. Results Seven compounds were isolated and obtained from salvia polyphenolic acids, which were identified as (2R)-2-[(3-(7-hydroxy-2-oxo-2,3-dihydrobenzofuran-4-yl)-(2E)-acryloyl) oxy]-3-(3,4-dihydroxyphenyl)-propanoic acid (1), salvianolic acid D (2), protocatechuic aldehyde (3), rosmarinic acid (4), lithospermic acid (5), salvianolic acid B (6), and salvianolic acid Y (7). Conclusion Compound 1 is a new compound, which is named as salvianolic acid D lactone.
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Objective To investigate the chemical constituents from the ethyl acetate extract of Gleditsiae Spina. Methods The chemical constituents were isolated and identified by chromatography on silica gel, Toyopearl HW-40C, Sephadex LH-20, MCI Gel CHP-20, ODS, and RP-HPLC. Their structures were elucidated on the basis of physicochemical properties and spectral analyses. Results Eighteen compounds were isolated from the ethyl acetate extract of Gleditsiae Spina, and identified as scopoletin (1), (-)-(7R,8S)-erythro-guaiacylglycerol (2), 5-methoxy-guaiacylglycerol (3), (-)-(7R,8R)-threo-guaiacylglycerol (4), 3,4’- dihydroxypropiophenone (5), dihydroferulic acid (6), p-hydroxybenzoic acid (7), indole-3-carbaldehyde (8), protocatechuic acid methyl (9), protocatechuic aldehyde (10), syringic acid (11), 2-guaiacylpropane-1,3-diol (12), (2R*,3R*,4S*)-2,3-diguaiacyl-4- hydroxyl tetrahydrofuran (13), 3-(2-oxopropyl)-3-hydroxy-indolin-2-one (14), juglanin D (15), C-veratroylglycol (16), 3-(4-hydroxyl-3-methoxyphenyl)-propan-1,2-diol (17), and (-)-syringaresinol (18). Conclusion Eighteen compounds are isolated from this plant for the first time.
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Objective To investigate the vasodilating effects of protocatechuic aldehyde (PCA ) on rat superior mesenteric arterial rings as well as its mechanism .Methods The tension of rat superior mesenteric arterial rings was recorded by a sensitive myograph system in vitro . We measured isometric tension changes in preconstricted rat superior mesenteric artery rings induced by potassium chloride (KCl ,60 mmol/L) ,serotomin (5-HT ,10-5 mol/L ) , and phenylephrine (PE , 10-5 mol/L ) after PCA treatment at different concentrations , respectively .We also observed vasodilating effects of PCA on KCl (60 mmol/L ) preconstricted rat superior mesenteric arterial rings after incubation with different inhibitors ,i .e .,L-NAME ,Indo ,ODQ ,4-AP (KV channel blocker) ,TEA (KCa channel blocker) ,BaCl2 (Kir channel blocker) ,and Glib (KATP channel blocker) ,respectively . Results PCA (10-6 -10-3 mol/L ) could relax KCl (60 mmol/L ) and 5-HT (10-5 mol/L ) preconstricted rat superior mesenteric arterial rings in a concentration-dependent manner . Indo of endothelial mechanism inhibitor blocked the vasodilating effect of PCA . 4-AP and BaCl2 of potassium ion channel inhibitors affected the vasodilatation induced by PCA in KCl (60 mmol/L)-preconstricted rat mesenteric artery .Conclusion PCA can relax KCl (60 mmol/L) ,or 5-HT preconstricted rat superior mesenteric arterial rings .This effect is associated with the inhibition of potassium channels and endothelial mechanism .
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OBJECTIVE: To investigate the chemical constituents of the seeds of Lepidium apetalum Willd. METHODS: The compounds were isolated and purified by Diaion HP-20, Toyopearl HW-40, MCI Gel CHP-20, ODS, silica gel chromatography combined with Pre-HPLC and the structures were identified on the basis of spectral data and physiochemical properties. RESULTS: Sixteen compounds were isolated and identified from the water extract as catechol(1), protocatechuic aldehyde(2), 2-phenyl acetamide(3), methyl-5-hydroxypyridine-2-carboxlate(4), benzylcarbamic acid(5), N-benzylacetamide(6), raphanuside C(7), 1-phenyl-1, 2-ethanediol(8), 2-(4-hydroxyphenyl)-ethanol(9), isorhamnetin-7-O-α-L-rhamnopyranoside(10), kaempferol(11), methyl 2, 4, 6-trihydroxybenzoate(12), 2-(4-hydroxyphenyl)acetonitrile(13), syringic acid(14), protocatechuic acid(15), and methyl sinapate(16). CONCLUSION: Compounds 1-16 are isolated from this plant for the first time.
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Objective: To study the chemical constituents of Periploca Radix. Methods: The chemical constituents of Periploca forrestii were separated using various chromatographic techniques. Their structures were elucidated by spectral analysis. Results: Nineteen compounds were isolated from the 70% ethanol extract of P. forrestii, and identified as 3-O-caffeoylquinic acid methyl ester (1), 4-O-caffeoylquinic acid methyl ester (2), 5-O-caffeoylquinic acid methyl ester (3), 3-O-caffeoylquinic acid (4), 4-O-caffeoylquinic acid (5), 5-O-caffeoylquinic acid (6), 1, 3-di-O-caffeoylquinic acid (7), 3, 4-di-O-caffeoylquinic acid (8), 3, 5-di-O- caffeoylquinic acid (9), 4, 5-di-O-caffeoylquinic acid (10), protocatechuic aldehyde (11), p-hydroxybenzoic acid (12), o-hydroxybenzoic acid (13), syringic acid (14), vanillic acid (15), periforgenin A (16), Δ5-pregnene-3β, 17α, 20α-triol (17), periforgenin C (18), and periplogenin (19). Conclusion: Compounds 1-12 are isolated from the plants of genus Periploca Linn. for the first time, and compound 13 is isolated from P. forrestii for the first time.
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OBJECTIVE:To establish a method for simultaneous determination of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride in Shenbai shuxin granules.METHODS:HPLC method was adopted.The separation was performed on Waters sunfire-C18 column with mobile phase consisted of acetonitrile-0.05 ％ trifluoroacetic acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 288 nm,and the column temperature was 30 ℃.The sample size was 5 μL.RESULTS:The linear ranges of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride were 0.040 01-1.600 46 μg(r=0.999 9),0.013 84-0.553 7 μg(r=0.999 9),0.049 32-1.972 94 μg(r=0.999 6),0.014 46-0.578 6 μg(r=0.999 8).The limits of quantitation were 7.68,2.66,4.74,1.38 ng,and the limits of detection were 1.92,0.66,2.36,0.69 ng,respectively.RSDs of precision,stability and reproducibility tests were all lower than 2％.The recoveries were 98.846％-100.762％ (RSD=0.77％,n=6),96.632％-99.463％ (RSD=0.98％,n=6),98.541％-100.432％ (RSD=0.82 ％,n =6),98.607 ％-101.521％ (RSD =1.11％,n =6),respectively.CONCLUSIONS:The method is simple,accurate and precise.It can be used for 4 components in Shenbai shuxin granules.
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To study the pharmacokinetics-pharmacodynamics correlation of protocatechuic aldehyde and hydroxysafflor yellow A alone or their combination use in rats with hyperlipidemia. In this study, the hyperlipidemia model was established by intravenous injection of protocatechuic aldehyde (20 mg•kg⁻¹) and hydroxysafflor yellow A (12 mg•kg⁻¹). The HPLC-DAD method was applied to determine the plasma concentration of protocatechuic aldehyde and hydroxysafflor yellow A at different time points and draw the drug effect-time curve. Meanwhile, the platelet activating factors (PAF) and plasma a granule membrane protein (GMP-140) contents were determined at different time points to draw the time-effect curve. Then DAS 3.2.6 software was used to process the data, analyze their correlation, and compare the difference of pharmacokinetics and pharmacodynamics of protocatechuic aldehyde and hydroxysafflor yellow A in hyperlipidemia rats after alone or their combined application, so as to evaluate the effect of protocatechuic aldehyde and hydroxysafflor yellow A on hyperlipidemia rats. According to the result, the pharmacokinetics and pharmacodynamics process of protocatechuic aldehyde and hydroxysafflor yellow A in hyperlipidemia rats after alone or their combination were consistent to the three-compartment model. In model group, the plasma PAF and GMP-140 were significantly increased, and the PAF and GMP-140 in vivo contents were decreased in a certain time after treatment. The effects of protocatechuic aldehyde and hydroxysafflor yellow A against the pharmacodynamic action may be related with their level in vivo, and their plasma concentration was positively related to the PAF and GMP-140 contents. The pharmacodynamic indexes were better after the combined use of protocatechuic aldehyde and hydroxysafflor yellow A, with certain influence on each other in hyperlipidemia rats; at the same time, it also reflected the rationality of protocatechuic aldehyde and hydroxysafflor yellow A combined application.
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Objective: To research the chemical composition from the roots of Lonicera japonica and their anti-inflammatory activities. Methods: The compounds were isolated and purified by chromatography on silica gel column, Sephadex LH-20 column, preparative thin-layer, and semi-preparative HPLC, etc. Their structures were identified on the basis of spectroscopic data, and parts of the isolated compounds were tested for their anti-inflammatory activities against zebrafish. Results: Fourteen compounds were isolated and elucidated as 3,13-dihydroxystemodan-2-one (1), chrysophanol (2), palmarumycin CP2 (3), β-sitosterol (4), stigmasterol (5), stigmast-4,6,8(14), 22-tetraen-3-one (6), erythrinassinate D (7), lanosterin (8), 3-O-acetyloleanolic acid (9), protocatechuin aldehyde (10), daucosterol (11), (E)-3-(3,4-dihydroxybenzylidene)-5-(3,4-dihydroxyphenyl)-2(3H)-furanone (12), lomacarinoside B (13), and (2E,6S)-8-(α-L-arabinopyranosyl-(1″→6')-β-D-glucopyranosyloxy)-2,6-dimethyloct-2-eno-1,2″-lactone (14). Conclusion: Compound 1 is a new compound, and compounds 2, 3, 6, 7-9, and 13 are isolated from L. japonica for the first time. Compounds 9 and 10 show the significant anti-inflammatory activities at 100 μg/mL.
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Objective: To develop a method for the quantitative determination of several salvianolic acids by HPLC and investigate the influence of danshensu and protocatechuic aldehyde on the degradation process of salvianolic acid B (SaB). Methods: HPLC method was applied to quantitative analysis on four components in the degradation sample of SaB. HPLC-MS method was applied to tentatively characterizing the structure of the degradation products. The effect of danshensu and protocatechuic aldehyde on the degradation of SaB were found out by the analysis of chromatogram and mass spectra of degradation samples. Results: Nine degradation products were found in the degradation samples of SaB, including danshensu, protocatechuic aldehyde, isomers of SaB, lithospermic acid, and its isomers. Protocatechuic aldehyde had stronger protective effect on SaB compared to danshensu at the same mass concentration. Additionally, the peak area of lithospermic acid isomers I and II increased when the initial concentration of protocatechuic aldehyde was increased. Conclusion The mild acidity of protocatechuic aldehyde might account for its stronger protective effect on SaB, and the connection between protocatechuic aldehyde and lithospermic acid isomers I and II might relate to the change of reaction balance.
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Objective: Using total quantum statistical moment analysis to evaluate the influence of the main effective constituents in Danhong Injection on the total pharmacokinetic parameters after orthogonal design to research internal compatibility rule of traditional Chinese medicine (TCM) compound. Methods: Using modified middle cerebral artery occlusion (MCAO) method to establish rat disease model of focal cerebral ischemia-reperfusion (I/R) injury. According to the orthogonal design L9 (34) composed of the main effective constituents (danshensu, protocatechuic aldehyde, salvianolic acid B, and HYSA) of Danhong Injection, nine combinations with different dosage ratios were formed, which were used for iv administration in MCAO rats to study the pharmacokinetics of effective constituents after compatibility. Using non-compartmental model of DAS 3.2.6 software to fit the pharmacokinetic parameters and using total quantum statistical moment analysis method to calculate the main total quantum statistical moment parameters. The influence of different compatibilities on the total pharmacokinetics parameters was analyzed by orthogonal test. Results: There were different effects of the main effective constituents with orthogonal compatibilities on the total quantum statistical moment parameters. Danshensu had the strongest effect on the total AUC and had a significant effect (P < 0.05). A3B3C2D1 was the preferred dose combination for AUCt. HYSA had the strongest effect on the total MRT and its optimal ingredient composition was A1B3C1D1. Conclusion: The total quantum statistical moment analysis could combine the pharmacokinetics of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and HYSA, and express the total pharmacokinetic behaviors. The total quantum statistical moment analysis method can be used to study the pharmacokinetics of multi-component in TCM compound prescription, which can provide the method for research on internal compatibility rule for TCM compound.
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OBJECTIVE:To establish a method for simultaneous determination of danshensu,salvianolic acid B,protocatechu-ic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule. METHODS:HPLC was performed on the column of Zorbax Eclipse XDB-C18 with mobile phase of acetonitrile-methanol-0.5% H3PO4(gradient elution)at a flow rate of 1.0 ml/min,the detec-tion wavelength was 280 nm(for danshensu,protocatechuic aldehyde,salvianolic acid B),230 nm(for ferulic acid)and 320 nm (for paeoniflorin),column temperature was 30 ℃,injection volume was 10 μl. RESULTS:The linear range was 1.19-478.34 μg/ml for danshensu(r=0.999 9),0.11-44.93 μg/ml for protocatechuic aldehyde(r=0.999 9),7.49-995.20 μg/ml for salvianolic acid B (r=0.999 7),0.95-379.39 μg/ml for paeoniflorin (r=0.999 9) and 0.01-3.12 μg/ml for ferulic acid (r=0.999 5);the limits of quantitation were 1.91 ng,0.36 ng,150.00 ng,2.74 ng and 0.10 ng,limit of detection were 0.96 ng,0.10 ng,45.00 ng,1.52 ng and 0.03 ng;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 98.06%-99.47%(RSD=0.52%,n=6),98.01%-99.49%(RSD=0.70%,n=6),98.44%-99.45%(RSD=0.37%,n=6),96.94%-100.71%(RSD=1.27%,n=6)and 95.44%-100.44%(RSD=1.90%,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the simultaneous determination of danshensu,salvianolic acid B,protocatechuic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule.
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Objective:To establish the quality control methods for Xinmeng Anshen granule. Methods:TLC was used to identify Salviae Miltiorrhizae Radix Et Rhizoma,Mori Fructus, Ziziphi Spinosae Semen( fried) ,Arachis Hypogaea L. and Schisandrae Chinensis Fructus in the granules. HPLC was used to determine the content of protocatechuic aldehyde in Salviae Miltiorrhizae Radix Et Rhizoma and spinosin in Ziziphi Spinosae Semen. Results:The TLC spots were clear and well-separated without any interference from the nega-tive sample. The calibration curves were linear within the range of 3.972-198.600 μg·ml-1(r=0.999 9) for protocatechuic alde-hyde and 7. 070-226. 240 μg·ml-1(r=0. 999 9) for spinosin. The average recovery was 98. 46% (RSD=1. 18%, n=9) for proto-catechuic aldehyde and 98. 20% (RSD=0. 90%, n=9) for spinosin. Conclusion:The established quality control methods are accu-rate, simple, repeatable and specific. It can be well used for the quality control of Xinmeng Anshen granule.
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Objective To explore the effect of heating temperature and heating time on the dissolution characteristics of phenolic acids in water extract of Danshen Decoction. Methods The contents of tanshinol, protocatechuic aldehyde and salvianolic acid B in water extract of Danshen Decoction were determined by reverse-phase high performance liquid chromatography (RP-HPLC) . The changes of the contents were monitored by classic constant temperature acceleration method. Results Of the three components, the content of salvianolic acid was decreased with the increase of heating temperature and the prolongation of heating time, and the degradation characteristics of the salvianolic acid B met the first-order kinetics equation. Conclusion The water-soluble components have less stability in the water extracts of Danshen Decoction. Lignum Santali Albi and Fructus Amomi accelerate the degradation speed of Radix Salviae Miltiorrhizae, suggesting that we should pay attention to the influence of heating temperature and heating time on the content of salvianolic acid B in the extraction and concentration process.
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Objective: A reversed-phase HPLC method was established for the simultaneous determination of five hydrophilic and lipophilic components in the roots of Salvia miltiorrhiza. Methods: Hydrophilic components including danshensu, protocatechuic aldehyde, and salvianolic acid B, and lipophilic components such as cryptotanshinone and tanshinone IIA, were successfully separated on a Waters Symmetry C18 reverse-phase column (250 mm × 4.6 mm, 5 μm), with acetonitrile-0.5% phosphoric acid (gradient elution) as mobile phase, the detection wavelength was set at 281 nm with flow rate of 1.0 mL/min, and the column temperature was maintained at 30 °C. Results: The recovery of the method was in the range of 95.1%-102.5% and the precision was less than 3% for all five analytes. All the compounds showed good linearity (R2 > 0.9990) in a relatively wide concentration range. Therefore, this HPLC method demonstrated good reproducibility, stability, and accuracy in validation studies. Conclusions: Simultaneous quantification of the multiple components by HPLC would be a better strategy for the quality evaluation on the roots of S. miltiorrhiza.
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Objective: By optimizing the chromatographic conditions, to establish a method of multi-target content analysis, the components of danshensu, protocatechuic aldehyde, salvianolic acid B, puerarin, and berberine hydrochloride in Xiaokeping Tablets were simultaneously determined. Methods: Agilent Zorbax Eclipse XDB-C18 (150 mm × 4.6 mm, 5 μm) chromatographic separation column was used, mobile phase was composed of acetonitrile (A), methanol (B), and 0.5% H3PO4 aqueous solution (C) in a linear gradient elution mode (0 min: A 4%, C 96%; 7 min: A 4%, B 1%, C 95%; 20 min: A 10%, B 3%, C 87%; 30 min: A 15%, B 3%, C 82%; 45 min: A 25%, B 10%, C 65%; and 60 min: A 30%, B 10%, C 60%). The flow rate was 1.0 mL/min, the column temperature was set at 30 ℃, and injection volume was 10 μL. The detection wavelength was set at 280 nm for danshensu, protocatechuic aldehyde, and salvianolic acid B, 250 nm for puerarin, and 265 nm for berberine hydrochloride. Results: There was a good linear relationship over a rang of 33.02-528.41 μg/mL for danshensu, 1.44-22.46 μg/mL for protocatechuic aldehyde, 374.51-5992.20 μg/mL for salvianolic acid B, 84.86-1357.82 μg/mL for puerarin, and 312.50-5000.00 μg/mL for berberine hydrochloride, respectively. The average recovery rates were 98.24% (RSD = 0.26%), 98.37% (RSD = 1.01%), 98.86% (RSD = 0.50%), 98.93% (RSD = 0.68%), 99.04% (RSD = 0.29%), respectively. Conclusion: The separation effect is good, the operation is simple and convenient, so as to achieve the better quality control of the products by this method.