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Aim To investigate the effects of sinapine thiocyanate (ST) on the malignant biological behavior of cutaneous squamous cell carcinoma A431 and Colo-16 cells, and its mechanism. Methods The fibroblast cells were treated with 20 μmol · L
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OBJECTIVE：To stud y the effects of sinapine thiocyanate （ST） on the proliferation ，epithelial mesenchymal transformation（EMT）and metastasis of human cutaneous squamous cell carcinoma SCL- 1 cells，and to investigate its possible mechanism. METHODS ：Human cutaneous squamous cell carcinoma SCL- 1 cells were divided into blank control group （0.1％ DMSO） and ST different concentration groups （5，10，20 μmol/L）. CCK- 8 assay，5-ethynyl-2′-deoxyuridine（EDU）test， scratch test and Transwell chamber invasion test were adopted to test the proliferation ，migration and invasion ability. The expression of N-cadherin and E-cadherin were detected by Western blot and immunofluorescence assay . Other SCL- 1 cells were collected and divided into blank control group （0.1％ DMSO），ST group （20 μmol/L），ST+NSC228155 group [ 20 μmol/L ST+100 μmol/L NSC228155（EGFR agonist ）] and ST+SC 79 group [ 20 μmol/L ST+20 μmol/L SC79（PI3K/Akt agonist ）]. The proliferation ，migration and invasion ability of SCL- 1 cells in each group were detected by CCK- 8 assay，scratch test and Transwell chamber invasion assay. The expression of epidermal growth factor receptor （EGFR），phosphatidylinositol 3 kinase（PI3K），phosphorylated phosphatidylinositol 3 kinase（p-PI3k），protein kinase B （Akt）and phosphorylated protein Akt （p-Akt）protein of cells in blank control group and ST different concentration groups（5，10，20 μmol/L）were determined by Western blot assay so as to validate the relationship between ST effect and EGFR/ PI3K/Akt signaling pathway. SCL- 1 cells and human normal skin fibroblasts cell WS 1 were divided into blank control group （0.1％ DMSO），ST group （20 μmol//L），ZD1839 group（positive control ，20 μmol//L，EGFR inhibitor ）and LY 294002 group（positive control，20 μmol//L，PI3K/Akt inhibitor ）. CCK- 8 assay was used to detect the cell proliferation in order to evaluate the cells cytotoxicity of ST. RESULTS ：Compared with blank control group ，the proliferation ，migration and invasion ability of SCL- 1 cells were significantly decreased in 5，10，20 μmol/L ST groups（P＜0.05）. Western blot and immunofluorescence assay showed that the expression of N-cadherin in SCL- 1 cells were decreased significantly in 5，10，20 μmol/L ST groups（P＜0.05），while the protein expression of E-cadherin was increased significantly （P＜0.05）；the protein expressions of EGFR ，p-PI3K and p-Akt were significantly decreased （P＜0.05）. Compared with ST group ，the proliferation ，migration and invasion ability of SCL- 1 cells were increased significantly in ST + NSC 228155 group and ST + SC 79 group （P＜0.05）. Compared with blank control group ，the proliferation ability of WS 1 cells had no significant change in ST group ，while the proliferation ability of SCL- 1 cells was decreased significantly （P＜0.05）；the proliferation ability of the two kinds of cells were decreased significantly in ZD 1839 group and LY 294002 group（P＜0.05）. Compared with ST group ，the proliferation ability of WS 1 cells was decreased significantly in ZD1839 group and LY 294002 group（P＜0.05），but there was no significant difference in the proliferation ability of SCL- 1 cells （P＞0.05）. CONCLUSIONS ：ST may inhibit the proliferation ，EMT and metastasis of SCL- 1 cells through inhibiting the activation of EGFR/PI 3K/Akt signaling pathway ，and its side effects are few.
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OBJECTIVE：To prov ide reference for the improvement of quality standards of Shiwei yipi granules. METHODS ： According to the general rules of 2015 edition of Chinese Pharmacopeia （part Ⅳ），microscopic identification was used to identify Massa Medicata Fermentata and Galli Gigerii Endothelium Corneum ；TLC method was used to qualitatively identify Crataegi Fructus and Semen Raphani ；the content of sinapine thiocyanate in Semen Raphani was determined by HPLC. RESULTS ：The microscopic characteristics were obvious for Massa Medicata Fermentata （palisade cells of testa and stone cells of testa ）and Galli Gigerii Endothelium Corneum （irregular fragments ）. The same fluorescent spots of Crataegi Fructus and Semen Raphani were displayed at the same position of ursolic acid ，sinapine thiocyanate control and Semen Raphani reference substance. The linear range of sinapine thiocyanate was 23.27-9 574.42 ng （r＝1.000 0）. The LOD and LOQ were of 0.50 μ g/mL and 1.68 μ g/mL， respectively. RSDs of precision ，repeatability，intermediate precision and stability tests （24 h）were all less than 1.5％. The average recoveries were 99.40％-100.89％（RSDs were 0.18％-0.49％，n＝3）. The contents of sinapine thiocyanate in 3 batches of Shiwei yipi granules were 0.086 4-0.220 6 mg/g，the average was 0.168 4 mg/g. CONCLUSIONS ：The identification method of Massa Medicata Fermentata ，Galli Gigerii Endothelium Corneum ，Crataegi Fructus and Semen Raphani in Shiwei yipi granules as well as the method for content determination of sinapine thiocyanate in Semen Raphani are established successfully. The content of Semen Raphani in the Shiwei yipi granules is no less than 0.16 mg/g calculated by sinapine thiocyanate （C16H24NO·5 SCN）.
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Objective: To analyze the dynamic changes of components in the enzymolysis process of raw products of Raphani Semen. Method: HPLC was employed to analysis of characteristic spectra of Raphani Semen at different enzymolysis time with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 225 nm.The characteristic peaks were calibrated, meanwhile, the UV spectra of characterstic peaks were extracted, and the difference between UV spectra and the changes of peak areas were compared, and the dynamic changes of characterstic components in Raphani Semen were analyzed. Result: Eleven characteristic peaks were marked from the characteric spectra of raw products of Raphani Semen at different enzymolysis time, and glucoraphenin and sinapine thiocyanate were assigned.Glucoraphenin was enzymatically hydrolyzed fastly by myrosinase, and an intermediate was generated, and then continue to be decomposed into other components.Sinapine thiocyanate did not change significantly during the enzymolysis process, and sinadiosides was also enzymatically degraded. Conclusion: The enzymolysis of Raphani Semen is not only the glucoraphenin, but also the sinadiosides.This paper can provide reference for the property change of Raphani Semen in processing.
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Objective: To compare the effect of four kinds of decocting containers on the content of sinapine and the HPLC specific chromatograms of Sinapis Semen decoction,so as to optimize decocting container for the development of classical formulas. Method: Selecting four kinds of decoction vessels,named traditional casserole,ceramic pot,round-bottom flask and stainless-steel pot as the research object,the content of sinapine in Sinapis Semen decoction and its HPLC specific chromatograms were used as indexes to investigate the influence of different decoction vessels on the decoction.Similarity evaluation of specific chromatograms was performed by the "Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine"(edition of 2004A). Result: The contents of sinapine in the decoction prepared by traditional casserole,ceramic pot,round-bottom flask and stainless-steel pot were 0.04%,0.07%,0.84% and 0.97%,respectively.Compared with specific chromatograms of the decoction prepared by traditional casserole,the similarities of specific chromatograms of the decoction prepared by ceramic pot,round-bottom flask and stainless-steel pot were 0.98,0.82 and 0.68,respectively.Compared with specific chromatograms of the decoction prepared by ceramic pot,the similarities of specific chromatograms of the decoction prepared by round-bottom flask and stainless-steel pot were 0.79 and 0.62,respectively.Compared with specific chromatograms of the decoction prepared by round-bottom flask,the similarity of specific chromatograms of the decoction prepared by stainless-steel pot was 0.97. Conclusion: The content of sinapine and HPLC specific chromatograms of Sinapis Semen decoction obtained from different decocting containers are quite different.
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OBJECTIVE： To establish a method for the content determination of sinapine thiocyanate， quercetin and kaempferol in rat plasma， and to study pharmacokinetics of Qili qiangxin capsule in rats in vivo. METHODS： HPLC-MS/MS method was adopted. The determination was performed on ZOBRAX XDB-C18 column with mobile phase consisted of 0.1％ formic acid solution and acetonitrile containing 0.1％ formic acid （gradient elution） at the flow rate of 0.45 mL/min. The sample size was 10 μL. Quantitative ions were sinapine thiocyanate with m/z 310.2→251.2， quercetin with m/z 301.1→150.7， kaempferol with m/z 286.2→242.0， internal standard chloramphenicol with m/z 320.9→ 151.9. 0.083， 0.167， 0.333， 0.667， 1， 1.5， 2， 3， 4， 6， 8， 12， 24 h after intragastric administration of Qili qiangxin capsule 1.3 g/kg， blood samples were collected via intraocular canthal venous plexus of 6 rats. The blood concentrations of sinapine thiocyanate， quercetin and kaempferol were determined. The pharmacokinetic parameters were calculated and fitted by using DAS 3.0 software. RESULTS： The linear range of sinapine thiocyanate， quercetin and kaempferol 0.05-100， 0.1-200， 0.1-200 ng/mL（r＝0.999 4， 0.999 7， 0.999 9）； RSDs of precision test and matrix effect were all less than or equal to 11.55％ （n＝6）， RE of stability test is less than or equal to 14.69％ （n＝3）. The pharmacokinetic parameter of sinapine thiocyanate， quercetin and kaempferol included that cmax were（1.35±0.62），（3.23±1.26），（5.27±1.66） ng/mL； tmax were （1.50±0.00）， （0.67±0.00）， （0.67±0.00） h； t1/2 were （3.98±0.99），（3.33±0.41），（4.54±0.85） h； CL were （3 683.82±987.96）， （2 852.33±695.88），（1 611.85±129.59） mL/（h·kg）； AUC0-24 h were （3.98±1.21）， （10.96±3.42）， （13.59±5.35） h·ng/mL. CONCLUSIONS： Established method is highly sensitive， specific and reproducible， and suitable for the pharmacokinetic study of sinapine thiocyanate， quercetin and kaempferol in rat.
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OBJECTIVE： To establish the fingerprint of Maizao yishen granules， and to provide scientific basis for its further development. METHODS： HPLC method was adopted to establish the fingerprint by using 10 batches of Maizao yishen granules sa samples. The determination was performed on Venusil XBP C18（L） column with mobile phase consisted of acetonitrile-0.2％ phosphoric acid （gradient elution） at the flow rate of 1→0.7 mL/min at 7-10 min， 0.7→1 mL/min at 10-15 min and 1 mL/min at the rest of time. The detection wavelengths were set at 284 nm （0-7 min）， 330 nm （7-32 min） and 360 nm （32-45 min）. The column temperature was 25 ℃， and sample size was 10 μL. The fingerprint of Maizao yishen granules was established， and the similarity evaluation was performed by using “Similarity Evaluation System of TCM Chromatographic Fingerprints” （2004 A edition） software. Then， the common peaks were assigned and identified by comparing reference substance and control medicinal materials. RESULTS： The precision， stability （24 h） and repeatability of the methodological investigation were all good [RSD values of relative retention time and relative peak area of each chromatographic peak were less than 3％ （n＝6）]. The similarity of 10 batches of samples were all above 0.900. Seventeen common peaks were identified， of which common peak 1 and 6 came from Semen Raphani； common peak 7， 9， 14， 15 and 16 from Citrus reticulata； common peak 5， 10， 11， 12 and 13 came from Glycyrrhiza uralensis； common peak 2 came from C. reticulata， G. uralensis and Ziziphus jujuba； peak 3 came from G. uralensis and Semen Raphani； peak 8 came from Hordeum vulgare and Semen Raphani； peak 4 and 17 came from C. reticulata and G. uralensis. Peak 1 was identified as hesperidin and the peak 9 was identified as sinapine. CONCLUSIONS： Established fingerprint of Maizao yishen granules is accurate and reliable， and can be used for quality control of Maizao yishen granules.
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Objective To study the mechanism of enhanced HaCaT cellular uptake of tetrahydropalmatine (THP) by asarum essential oil (AEO) and sinapine thiocyanate (SPT) in Sanfu Patch. Methods Effect of SPT, AEO, and THP on cell viability of HaCaT were determined by MTT. HaCaT cellular uptake of THP and the enhancing effects of AEO and SPT on THP uptake were visualized with confocal laser scanning microscope (CLSM) based on the green autofluorescence of THP, and the THP uptake content by HaCaT was further determined with HPLC. Moreover, HaCaT cells were labeled with diphenylhexatriene (DPH). After the labeled cells were treated with AEO, SPT, and THP, respectively, the cellular membrane fluidity was determined with fluorescence polarization technology. Results THP fluorescence intensity within HaCaT cells was significantly increased when THP was co-delivered with AEO or SPT respectively, and the THP content with each group within the cells was also significantly higher than that of THP delivered alone. In addition, AEO was superior to SPT in enhancing THP uptake by HaCaT cells. The fluorescence polarization and membrane micro-viscosity of HaCaT cells were significantly decreased after AEO treatment, which indicated that membrane fluidity was increased by the treatment with AEO. However, SPT or THP did not present the character of increasing the membrane fluidity.Conclusion HaCaT cellular uptake of THP can be enhanced by AEO and SPT of Sanfu Patch, in which AEO enhances the cellular uptake of THP through increasing the cellular membrane fluidity, while the mechanism of SPT in enhancing THP cellular uptake remains further clarification.
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AIM:To investigate the effects of sinapine,an effective monomer of Chinese medicine,on hydro-gen peroxide(H2O2)-induced adipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).METHODS:The undifferentiated rat BMSCs were identified and screened by flow cytometry.The adipogenic differentiation of BMSCs was induced by H2O2,and the toxicity of sinapine on BMSCs was tested by CCK-8 assay.After the modeling method and the concentration range of sinapine were determined,the lipid droplets in the cells were detected by Oil Red O semi-quanti-tative assay,and the optimal drug concentration was selected.Finally,Oil Red O assay was observed 24 h after drug inter-vention,and the expression of adipogenic differentiation-related proteins,adipocyte protein 2(aP2),peroxisome prolifera-tor-activated receptor γ(PPARγ)and glucose transporter 4(Glut4),at mRNA and protein levels in the BMSCs was deter-mined by qPCR and Western blot.RESULTS:Treatment with H2O2at 200 μmol/L for 1 h induced BMSCs to differentiate into adipocytes.Below the concentration of 40 μmol/L,sinapine had no toxicity to BMSCs.The best inhibitory concentra-tion of sinapine on adipogenic differentiation was at 15 μmol/L.The number of lipid droplets in sinapine(15 μmol/L) group was significantly lower than that in model group.In sinapine group,the expression of aP2,PPARγand Glut4 at mR-NA and protein levels was lower than that in model group(P<0.01).CONCLUSION: Sinapine inhibits H2O2-induced adipogenic differentiation of rat BMSCs.The mechanism may be related to the PPARγ/AMPK signaling pathway.
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AIM:To investigate the inhibitory effect of sinapine thiocyanate(ST)on hyperglycemia,hyper-lipemia,atherosclerosis and hepatocellular steatosis of ApoE-/-mice with insulin resistance(IR)and the possible mecha-nisms.METHODS:ApoE-/-male mice(n=60)were assigned randomly into control group ,saline group,rosiglitazone group and ST treatment groups(at low,middle and high doses )with 10 mice in each group.The mice in control group were fed with fundamental diet ,while the mice in other groups were fed with high-fat diet for 12 weeks.The mice in ST groups were given gavage with different doses of ST(10,30 and 90 mg· kg-1· d-1)simultaneously,while the mice in rosiglitazone group received gavage with rosigltazone(1.33 mg· kg-1 · d-1 ).In the last 3 weeks,the mice in control group received daily intrape-ritoneal injection of physiological saline ,and IR was induced in other groups by daily intrape-ritoneal injection of dexamethasone(0.8 mg/kg).The blood sample was collected and fasting plasma glucose was tested weekly through tail vein.After all animals fasted for 12 h at the end of the 12th week,they were sacrificed and the levels of fasting insulin,tumor necrosis factor-α(TNF-α),triglyceride,total cholesterol and liver lipids were measured.The li-ver tissue and aortic immobilized sections were detected by HE staining.The expression of the proteins related to liver lipid metabolism and skeletal muscle MAPK signaling pathway was determined by Western blot.RESULTS:ST showed dose-dependently reduced serum lipids ,plasma glucose and TNF-α(P<0.05),delayed hepatocellular steatosis and atheroscle-rosis,and dose-dependently regulated hepatic lipid metabolism signaling molecules(HMGR and SREBP-2)and MAPK signaling molecules(ERK and p38)(P<0.05).CONCLUSION:ST has the biological potential of reducing blood li-pids and relieving IR.The mechanism may be related to the regulation of liver lipid metabolism and skeletal muscle MAPK signaling pathway.
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Objective The quality evaluation of the Shuwei mixture was determined by the content of the six components of sinapine cyanide sulfonate, magnolol, honokiol, hesperidin, naringin and neohesperidin. Methods RP-HPLC method was used.The separation was performed on a Agilent ZORBAX Eclispse SB-C18column (4.6 mm×250 mm,5 mm),the mobile phase consisted of acetonitrile(A)-0.1% phosphoric acid with gradient elution at the flow rate of 1.0 mL·min-1.The detection wavelength was 326 nm ( sinapine cyanide sulfonate ), 294 nm ( magnolol, honokiol ) and 283 nm ( naringin, neohesperidin).The column temperature was kept at 30 ℃. Results The sinapine cyanide sulfonate, magnolol, honokiol, hesperidin,naringin and neohesperidin all had good linear relationship in the ranges of 0.049 6-1.24,0.048 2-1.205,0.060 5-1.512 5,0.187 2-4.68,0.131 6-3.29,0.197-4.925 μg.The average recoveries were 100.66%, 99.86%, 101.37%, 102.41%, 99.01%, 102.05%, respectively, RSD were 0.82%, 1.89%, 2.56 %, 0.74%, 1.54%, 0.99%, respectively. Conclusion The method is simple,accurate,reproducible and nice to the separation,and can be used for the quality evaluation of Shuwei mixture.
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Objective To investigate and optimize the penetration enhancers of Xiaochuan emplastrum. Methods Franz diffusion cell was employed with isolated mice abdominal skin as barrier. The accumulating osmotic quantity per unit area and penetrating absorption rate of Sinapine thiocyanate were determinated by HPLC method. The penetration enhancers were selected and the ratio was determined. Results Combination of azone and propylene glycol were better than other kinds of enhancers for penetration effect, the ratio of propylene glycol and azone was 2:1. Dosage of azone and propylene glycol was finally optimized to 5% of prescription dosage.Under these conditions, cumulative permeation quantity of Sinapine thiocyanate was 369.59 μg·(cm2)-1, penetration rate of Sinapine thiocyanate was 14.19 μg·(cm2)-1·h-1 and enhancing rate was 3.19. Conclusion The ratio of propylene glycol and azone was 2:1,dosage of azone and propylene glycol was 5% of prescription dosage, which has a significant penetration effect and can be used as the penetration enhancer of Xiaochuan emplastrum.
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Objective To investigate and optimize the penetration enhancers of Xiaochuan emplastrum. Methods Franz diffusion cell was employed with isolated mice abdominal skin as barrier. The accumulating osmotic quantity per unit area and penetrating absorption rate of Sinapine thiocyanate were determinated by HPLC method. The penetration enhancers were selected and the ratio was determined. Results Combination of azone and propylene glycol were better than other kinds of enhancers for penetration effect, the ratio of propylene glycol and azone was 2:1. Dosage of azone and propylene glycol was finally optimized to 5% of prescription dosage.Under these conditions, cumulative permeation quantity of Sinapine thiocyanate was 369.59 μg·(cm2)-1, penetration rate of Sinapine thiocyanate was 14.19 μg·(cm2)-1·h-1 and enhancing rate was 3.19. Conclusion The ratio of propylene glycol and azone was 2:1,dosage of azone and propylene glycol was 5% of prescription dosage, which has a significant penetration effect and can be used as the penetration enhancer of Xiaochuan emplastrum.
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Objective:To study the transdermal absorption in vitro of sinapine thiocyanate,tetrahydropalmatine and asarinin in Xiaochuan ointment so as to offer reference for its clinical application.Methods:A Franz diffusion cells method with isolated rat skins was carried out to study the percutaneous rate of sinapine thiocyanate,tetrahydropalmatine and asarinin determined by LC-MS/MS.Results:With the increase of administration dosage,the cumulative penetration of sinapine thiocyanate,tetrahydropalmatine and asarinin showed few changes.The results showed that the transdermal behavior of sinapine thiocyanate fit to a Higuchi equation,and that of tetrahydropalmatine and asarinin fit zero-order process.The penetration rate of tetrahydropalmatine and asarinin respectively was 0.362×10-1 and 0.330×10-2 μg·cm-2·h-1.Conclusion:Xiaochuan ointment exhibits transdermal penetration and absorption properties,which provides evidence for its transdermal administration.
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Objective:To establish the quality standard for Sinapis Semen( stir-baked) formula granule. Methods:TLC was used to identify Sinapis Semen; an HPLC method was applied in the content determination of Sinapine thiocyanate in Sinapis Semen ( stir-baked) formula granule with Hibar Purospher STAR C18 (250 mm × 4. 6 mm,5 μm ) column and with the mobile phase consisting of acetonitrile-0. 08 mol·L-1 potassium dihydrogen phosphate (10 ∶90) at the flow rate of 1. 0 ml·min-1 ,the column temperature was 35℃ and the detection wavelength was set at 326 nm. Results:The characteristic spots of Sinapis Semen( stir-baked) formula granule were clearly detected by the established TLC chromatography. Sinapine thiocyanate had a good linear relationship within the concentra-tion range of 0. 012 2-3. 912 3μg(r=0. 999 9). The average recovery was 100. 4% (RSD=0. 6%, n=6). The water content, dis-solution and particle size of Sinapis Semen formula granule all met the related requirements. Conclusion:The methods are simple and accurate with good reproducibility,which can be used to control the quality of Sinapis Semen( stir-baked) formula granule.
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Objective: To compare the electric resistance, stratum corneum thickness, and biochemical composition in acupoint and non-acupoint skin, and the osmotic properties of sinapine by acupoint and non-acupoint administration with Baijiezi Tufang (Baijiezi liniment prescription), to investigate the effect of dermal biophysical characteristics on cutaneous administration of sinapine, and to provide the experimental support of acupoint transdermal of Baijiezi Tufang for the treatment of asthma. Methods: Skin electrical resistance was measured with a specially designed meter, the thickness of stratum corneum was detected through microscope after the skin slice was stained by HE, and the skin biochemistry ingredients were compared by laser Raman spectroscopy. The permeability through acupoint and non-acupoint skin was examined with the modified Franz diffusion cell in vitro and tape-stripping in vivo, using sinapine as index. The content of sinapine was determined by HPLC. In in vivo permeability experiment, the stratum corneum was isolated and the distribution of drugs in various layers of acupoint and non-acupoint was compared. Results: The dermal resistance in acupoint was lower than that in non-acupoint (P < 0.05) and the stratum corneum of acupoint was thinner than that of non-acupoint (P < 0.05). However, the biochemistry ingredients of them were almost the same. The 24 h cumulative amount through the acupoint skin was about 6.8 times as that through the non-acupoint skin, and the steady-state flux of sinapine across acupoint skin was 6.1 times as that across in non-acupoint skin. Sinapine retention in acupoint skin was significantly higher than that in non-acupoint skin (P < 0.05) both in vitro and in vivo after 24 h of administration, and the retention in stratum corenum of acupoint was also higher than that in stratum corneum of non-acupoint at different time points within 24 h. Conclusion: The permeation and retention of sinapine in Baijiezi Tufang through acupoint skin are higher than those through non-acupoint skin, which relates to the lower electrical resistance and thinner stratum corenum of acupoint skin. It suggests the rationale evidence for acupoint administration in clinic application of Baijiezi Tufang.
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Objective:To establish the quality standard of sinapine thiocyanate in Qingrehuashiyangyin granules by HPLC.Methods:HPLC was performed on Phenomenex C18 Column(250mm?4.6mm,5?m) with acetonitrile-0.08mol/L KH2PO4(15:85) as mobile phase.Flow rate was 1.0ml/min and detecting wave length was 326nm.Column temperature at 30℃.Results:The good linear relationship was obtained in the range of 0.3976--1.988?g(r=0.9998),and the average recovery was 99.0%.Conclusion:This method is simple,fast and accurate for the quality control of Qingrehuashiyangyin granules.
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Objective To develop an HPLC method for the assay of sinapine thiocyanate in Semen Sinapis Albae of different processed products. Methods It was assayed by RP-HPLC. The column was Hypersil BDS Phenyl (5 ?m, 250 mm?4.6 mm), the mobile phase was acetonitrile-0.08 mol/L KH2PO4 (15∶85). The UV detection wavelength was 240 nm. The flow rate was 1.0 mL/min. Results Good linearity of sinapine thiocyanate was showed within the range of 0.8～4.0 ?g (r=0.999 7). The average recovery was 100.98% (RSD=3.51%). Conclusion The method is simple, accurate and good reproducible. It can be used for quality control of Semen Sinapis Albae of different processed products.
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Objective To determine the content of Sinapine thiocyanate in the Baixi Kechuan Cream by HPLC and to lay foundation for establishing the quality control standard of the preparation. Methods The HPLC system consisted of the column of DIKMA inertsil (pH=3,5 ?m,250 mm?4.6 mm). The acetonitrile and 3% acetic acid were used as the mobile phase A and B,respectively,and gradient elution was used. The detective wavelength was 326 nm and the flow rate was 1.0 mL/min. The column temperature was room temperature. Results The method was linear within the range of 0.8～4.0 ?g (r=0.999 6,n=5). The average recovery was 100.83%,and RSD=1.95% (n=5). Conclusion The method is quick,simple,highly reproductive and steady,and it can be used for quality control of the preparation.
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Objective:Preparated from seeds of Brassica alba, Sinapine was determinated by RP-HPLC. Methods: Seeds of Brassica alba were shattered, defatted by ether, extracted by alcohol, and crystallized by potassium thiocyanate, then ethanol and water were used to recrystallize for several times. The determination was performed by reversed-phase high performance liquid chromatography(RP-HPLC), using ZORBAX SB-Aq column 4.6 mm?250 mm, 5 ?m. The mobile phase consisted of acetonitrile-0.08 mol/L KH2P04 (20:80) at a flow rate of 1.0 ml/min, and the detection wavelength was 326 nm. Results: Pale yellow needle-like sinapine crystal was obtained, and the yield rate was 0.82%. The purity of sinapine thiocyanate crystal was over 95%. Conclusion: Preparation sinapine by this solvent extraction method was simple, high-purity, high yield, and could be used for industrial production.