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During the analysis of benziamidazole-class irreversible proton pump inhibitors,an unusual mass spectral response with the mass-to-charge ratio at[M+10]+intrigued us,as it couldn't be assigned to any literature known relevant structure,intermediate or adduct ion.Moreover,this mysterious mass pattern of[M+10]+has been gradually observed by series of marketed proton pump inhibitors,viz.omeprazole,pantoprazole,lansoprazole and rabeprazole.All the previous attempts to isolate the corresponding component were unsuccessful.The investigation of present work addresses this kind of signal to a pyridinium thiocyanate mass spectral intermediate(10),which is the common fragment ion of series of labile aggregates.The origin of such aggregates can be traced to the reactive intermediates formed by acid-promoted degradation.These reactive intermediates tend to react with each other and give raise series of complicated aggregates systematically in a water/acetonitrile solution by electrospray ioniza-tion.The structure of the corresponding pyridinium thiocyanate species of omeprazole(10a)has been eventually characterized with the help of synthetic specimen(10a').Our structural proposal as well as its origin was supported by in situ nuclear magnetic resonance,chemical derivatization and colorimetric experiments.
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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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Cr(III) and Mn(II) metal complexes of Schiff base ligand derived from phenylacetylurea condensed with salicylaldehyde (SBPS) and thiocyanate(SCN-) ion were synthesized by using microwave irradiation. Microwave assisted synthesis gives high yield of the complexes within a very short time. The molecular formulae and the geometry of the complexes have been deduced from elemental analysis, metal estimation, electrical conductance, magnetic moment, electronic spectra, FT- IR, Far IR spectra, cyclic voltammetry, thermal analysis and powder-XRD techniques. The molar conductance values indicate that the complexes are non-electrolyte (1:0) type. FT-IR spectra show that Schiff base and thiocyanate ion are coordinated to the metal ion in a monodentate manner. The electronic spectra and the magnetic moment indicate the geometry of the complexes is found to be octahedral. The antimicrobial activities of ligands and their Cr(III) and Mn(II) complexes were studied against the microorganisms, viz., E. coli, Klebsiella Pneumonia, P. aeruginosa, S. aureus, Bacillus cereus, Aspergillus flavus, Aspergillus niger, Aspergillus oryzae, Aspergillus sojae and Candida albicans by agar well diffusion method. The complexes show moderate activity against the bacteria and enhanced activity against the fungi as compared to free SBPS ligand. The free radical scavenging activity of the complexes and the ligands has been determined by measuring their interaction with the stable free radical, DPPH. The complexes have larger antioxidant activity as compared to the ligand. DNA-binding properties have been studied by fluorescence-emissions method. The results suggest that the complexes strongly bind to DNA because of metal complexes are well-known to speed up the drug action and the ability of a therapeutic agent which can frequently be enhanced upon coordination with a metal ion.
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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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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 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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OBJECTIVE: To establish a method for detecting thiocyanate in human urine by high performance liquid chromatography( HPLC) with 2,3,4,5,6-pentafluorobenzyl bromide as precolumn derivatization reagent.METHODS: Thiocyanate in human urine was derived with 2,3,4,5,6-pentafluorobenzyl bromide, and separated by poroshell 120EC-C18 column with acetonitrile:deionized water( 60:40,V/V) as mobile phase.detected by HPLC,Liquid chromatography-UV detector was used for determination.The wavelength was 212.00 nm.RESULTS: Good linearity was obtained in the range of 0.05-10.32 mg/L with the correlation coefficient of 0.999.The detection limit was 6.31 μg/L and the minimum detection concentration was 63.10 μg/L( 0.1 mL urine).The recovery rate was 95.1%-102.9%.The within-run relative standard deviation( RSD) and the between-run RSD were 0.9%-1.0% and 0.9%-2.1%,respectively.The urine samples could be stored at 4 ℃ for 7 days.CONCLUSION: This method has high sensitivity,good specificity and sample preparation,which can be used for detecting urine thiocyanate in occupational population.
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OBJECTIVE: To establish a method for detecting human urinary thiocyanate by gas chromatographic and pre-column derivatization with 2,3,4,5,6-pentafluorobenzyl bromide( PFB-Br). METHODS: A total of 20. 0 μL of urine was taken and 1. 0 m L of acetonitrile and 100. 0 μL of PFB-Br were added for derivative reaction. The gas chromatography was directly used for measurement. RESULTS: The urinary thiocyanate concentration showed a good linear range of 1. 000-10. 000 mg/L. The linear correlation coefficient was 0. 999 6. The minimum detection concentration was 0. 112 mg/L,and the minimum quantitative concentration was 0. 411 mg/L( 20. 0 μL urine sample). The standard recovery rate was 97. 22%-102. 04%.The within-run relative standard deviation( RSD) of this method was 1. 56%-5. 35%. The between-run RSD was 1. 46%-5. 10%. Hydrocyanic acid ions interfered with the measurement. Other common inorganic ions such as chloride,sulfate,and nitrate ions did not interfere with the measurement results. The samples can be stored at 4 ℃ for at least 15 days. CONCLUSION: This method is suitable for detecting human urinary thiocyanate.
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Objective@#To establish ion chromatography method to determine thiocyanate within urine of workers who were exposed to cyanide.@*Methods@#After the workers work, used 50 ml centrifuge tube to collect the urine of workers about 20 ml.The urine were tested by centrifugation, dilution and filtration by C18 column, thiocyanate was separated by AS16 and mobile phase elution by KOH, detected by electrical circuitryconductivity detector, quantitative by the standard curve method.@*Results@#The linear correlation coefficient of thiocyanate within the range of 0.1-5.0 μg/ml was more than 0.999. The method detection limit was 0.11μg/ml (in 1ml urine) , the method quantitative limit was 0.35 μg/ml. The method recoveries were 95.1%-99.7%. The within-day precision range was 0.54%-2.05%, The between-run precision range was 2.06%-5.09%. Sample stability test showed that thiocyanate samples could be stored for 5 days at room temperature and 7 days at 4 ℃, could be stored for 14 days at-20 ℃.@*Conclusion@#The technical indicator of method compliance with rule of Guide for establishing occupation health standards-Part 5: determination methods of chemicals in biological materials (GBZ/T 210.5-2008) , the method applies to workers who were exposed to cyanide.
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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 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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Aims: To evaluate the levels of salivary thiocyanate and its relation with the occurrence of micronuclei (MN) using exfoliative cytology in smokers and nonsmokers. Materials and Methods: One hundred and twenty patients were divided into 3 groups: nonsmoker group 1 (control), smokers group 2, and smokers group 3. Their saliva was collected and analyzed for thiocyanate levels, and exfoliative cytology was evaluated for the presence of MN. Statistical Analysis Used: Fisher’s exact test and ANOVA test were used. Results: It was seen that as the grade of smoking increased, the levels of salivary thiocyanate and occurrence of MN increased. Conclusions: Detection and quantification of “biomarkers” such as salivary thiocyanate and MN in noninvasive and painless procedures such as oral exfoliative cytology can be an upcoming research domain in the field of cancer prevention and therapeutics.
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Aim: This study was conducted to estimate and correlate salivary thiocyanate (SCN) levels in periodontally healthy subjects, smokers, nonsmokers, and gutka‑chewers with chronic periodontitis. Methodology: The study population consisted of 40 systemically healthy subjects in the age group of 18–55 years that was further divided into four groups: Control, smokers, nonsmokers, and gutka‑chewers with chronic periodontitis. Gingival index (GI) (Loe and Silness‑1963), probing depth (PD), clinical attachment loss was assessed. Estimation of SCN was performed by ultraviolet spectrophotometer at 447 nm wavelength. Statistical analysis was performed using the one‑way ANOVAs Welch test and Pearson’s correlation test using SPSS version 17 software. Results: Results showed statistically significant increase in SCN levels in smokers as compared to gutka‑chewers with chronic periodontitis, control, and nonsmokers with chronic periodontitis subjects. Significantly higher PD and loss of attachment were seen in smokers group compared with other groups. A negative correlation observed between the GI and thiocyanate levels. Conclusion: The present study revealed a significant increase in SCN levels in smokers with periodontitis as compared to nonsmokers.
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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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In this present work two colorimetric methods were developed based on donor-acceptor complexes of alverine citrate (ALV) and tapentadol (TAP) with cobalt thiocyanate. These methods were developed on Perkin Elmer LAMBDA 25 UV–VIS spectrophotometer with 1cm quartz cells. The colored species formed are the coordination complexes of the drugs (electron donor) and the central metal atom of cobalt thiocyanate (electron acceptor) which is extractable into nitrobenzene from aqueous solution. The reaction conditions were optimized and validated to achieve maximum colour intensity. The colored complexes show maximum absorbance measured at 625 nm for both ALV and TAP. The absorbances were found to increase linearly with an increase in concentration which was corroborated by the calculated regression coefficients (0.9998-0.9999). Linearity was obeyed in the range of 100-600 μg/ml for both ALV and TAP, respectively. The molar absorptivity, sandell’s sensitivity, LOD, LOQ and other validation parameters have been evaluated extensively as per ICH guidelines and all the parameters were found within the acceptance criteria for both methods. The proposed methods were proven to be more accurate, simple, precise and rapid by statistical validation of recovery studies and could be suitable for regular analysis.