Dynamic regulation using a quorum-sensing circuit enhances the production of tyrosol by Escherichia coli / 生物工程学报

Tyrosol is a natural polyphenolic product that is widely used in chemical, pharmaceutical and food industries. Currently, the de novo synthesis of tyrosol by Escherichia coli suffers from issues such as low cell density and poor yield. Therefore, the phenylpyruvate decarboxylase mutant ARO10F138L/D218G obtained in our previous study was fused with an alcohol dehydrogenase from different microorganisms for fusion expression, and the optimal ARO10F138L/D218G-L-YahK produced 1.09 g/L tyrosol in shake flasks. In order to further improve tyrosol production, feaB, a key gene in the competing pathway of 4-hydroxyphenylacetic acid, was knocked out, and the resulted strain produced 1.26 g/L tyrosol with an increase of 21.15% compared to that of the control. To overcome the low cell density in tyrosol fermentation, the quorum-sensing circuit was used to dynamically regulate the tyrosol synthesis pathway, so as to alleviate the toxic effect of tyrosol on chassis cells and relieve the growth inhibition. Using this strategy, the yield of tyrosol was increased to 1.74 g/L, a 33.82% increase. In a 2 L fermenter, the production of tyrosol in the engineered strain TRFQ5 dynamically regulated by quorum-sensing reached 4.22 g/L with an OD600 of 42.88. Compared with those in the engineered strain TRF5 statically regulated by induced expression, the yield was increased by 38.58% and the OD600 was enhanced by 43.62%. The combination of blocking the competing pathway using gene knockout technology, and reducing the inhibitory effect of tyrosol toxicity on chassis cells through quorum-sensing dynamic regulation increased the production of tyrosol. This study may facilitate the biosynthesis of other chemicals with high toxicity.

Metabolic engineering of Escherichia coli for production of hydroxytyrosol / 生物工程学报

Hydroxytyrosol is an important fine chemical and is widely used in food and medicine as a natural antioxidant. Production of hydroxytyrosol through synthetic biology is of important significance. Here we cloned and functionally characterized a hydroxylase encoding gene HpaBC from Escherichia coli BL21, and both subunits of this enzyme can be successfully expressed to convert the tyrosol into hydroxytyrosol. A HpaBC gene integration expression cassette under the tac promoter was constructed, and integrated into the genome of a tyrosol hyper-producing E. coli YMG5A*R using CRISPR-Cas9 technology. Meanwhile, the pathway for production of acetic acid was deleted, resulting in a recombinant strain YMGRD1H1. Shake flask fermentation showed that strain YMGRD1H1 can directly use glucose to produce hydroxytyrosol, reaching a titer of 1.81 g/L, and nearly no by-products were detected. A titer of 2.95 g/L was achieved in a fed-batch fermentation conducted in a 5 L fermenter, which is the highest titer for the de novo synthesis of hydroxytyrosol from glucose reported to date. Production of hydroxytyrosol by engineered E. coli lays a foundation for further construction of hydroxytyrosol cell factories with industrial application potential, adding another example for microbial manufacturing of aromatic compounds.

Dynamic changes of five constituents in Ligustri lucidi Fructus at five picking time / 中成药

AIM To analyze the dynamic changes of five constituents in Ligustri lucidi Fructus at five picking time (August,September,October,November,December).METHODS The HPLC analysis of Ligustri lucidi Fructus ethanol extract was performed on a 25 ℃ thermostatic Aglient Zorbax SB-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 224 nm.RESULTS Salidroside,tyrosol,luteolin-7-O-glucoside,ligustroflavone and specnuezhenide showed good linear relationships within their own ranges (r ＞0.999 0),whose average recoveries were 99.56％-100.30％ with the RSDs of 0.89％-1.23％.The contents of various constituents (except for tyrosol) were the highest in samples picked up in September,followed by those picked up in October.CONCLUSION The suitable picking time of Ligustri lucidi Fructus is September and October.

Simultaneous determination of salidroside and tyrosol in Beagle dog plasma using UHPLC-MS/MS after pre-column dansyl chloride derivatization / 药学学报

A pre-column derivatization method combined with UHPLC-MS/MS was developed for the simultaneous determination of salidroside and tyrosol in Beagle dog plasma. After protein precipitation by acetonitrile, the liquid supernatant was treated with dansyl chloride under dark conditions at 60℃ for 30 min, and then, the sample solution was extracted using methyl tertiary butyl ether. The multiple reaction monitoring in positive ion mode was used for MS detection of the tested analytes with the specific ion transitions of m/z 534.2→372.0 for salidroside derivative, m/z 372.0→171.0 for tyrosol derivative and m/z 506.0→171.0 for arbutin derivative. The chromatograph separation was achieved on an ACQUITY UPLC® BEH C18 column (100 mm×2.1 mm, 1.7 μm) with a gradient mobile phase consisting of acetonitrile (0.1% formic acid)-water (10% acetonitrile, 0.1% formic acid) for 9 min. The assay showed a good linearity over the range of 0.02/0.1-20/10 μmol·L-1 with a lower limit of quantitation of 0.02 and 0.1 μmol·L-1 for salidroside and tyrosol in dog plasma, respectively. The intra-and inter-day precisions were all less than 8.68%, and the accuracy was within ±11.4%. The established method with a high sensitivity, good specificity and reliability was appropriate for simultaneous determination of salidroside and tyrosol in dog plasma and successfully applied to a pharmacokinetic study after intragastric administration of salidroside to Beagle dogs.

Research progress on vascular pharmacologic effect of salidroside / 药物评价研究

Salidroside antagonizes hypoxia,H2O2,hyper-glucose,homocysteine,smoking,orweightlessness-induced endothelial cell injury,and antagonizes the proliferation of vascular smooth muscle cells and vascular adventitial fibroblasts induced by hypoxia,hyper-glucose,noradrenaline,platelet-derived growth factor,or angiotensin Ⅱ,thus produces vascular protection and improves vascular function.Salidroside plays a dual role of vasoconstriction and vasodilation in regulating resistant blood vessel.The vasodilatory effect of salidroside is endothelium-dependent and endothelium-independent.Salidroside antagonizes vascular contraction and injury induced by KC1,CaC12,noradrenaline,phenylephrine,homocysteine,hyper-glucose,plateau hypoxia,H2O2,and chlorine.Salidroside relieves cerebral vasospasm induced by subarachnoid hemorrhage,and pulmonary hypertension and pulmonary arterial remodeling induced by hypoxia or monocrotaline in rat,attenuates several experimental atherosclerosis,and enhances plaque stability.

Research progress on vascular pharmacologic effect of salidroside / 药物评价研究

Salidroside antagonizes hypoxia,H2O2,hyper-glucose,homocysteine,smoking,orweightlessness-induced endothelial cell injury,and antagonizes the proliferation of vascular smooth muscle cells and vascular adventitial fibroblasts induced by hypoxia,hyper-glucose,noradrenaline,platelet-derived growth factor,or angiotensin Ⅱ,thus produces vascular protection and improves vascular function.Salidroside plays a dual role of vasoconstriction and vasodilation in regulating resistant blood vessel.The vasodilatory effect of salidroside is endothelium-dependent and endothelium-independent.Salidroside antagonizes vascular contraction and injury induced by KCl,CaCl2,noradrenaline,phenylephrine,homocysteine,hyper-glucose,plateau hypoxia,H2O2,and chlorine.Salidroside relieves cerebral vasospasm induced by subarachnoid hemorrhage,and pulmonary hypertension and pulmonary arterial remodeling induced by hypoxia or monocrotaline in rat,attenuates several experimental atherosclerosis,and enhances plaque stability.

Terpinen-4-ol, tyrosol, and ß-lapachone as potential antifungals against dimorphic fungi

Abstract This study aimed to evaluate the in vitro antifungal activity of terpinen-4-ol, tyrosol, and β-lapachone against strains of Coccidioides posadasii in filamentous phase (n = 22) and Histoplasma capsulatum in both filamentous (n = 40) and yeast phases (n = 13), using the broth dilution methods as described by the Clinical and Laboratory Standards Institute, to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of these compounds. The mechanisms of action of these compounds were also investigated by analyzing their effect on cell membrane permeability and ergosterol synthesis. The MIC and MFCf these compounds against C. posadasii, mycelial H. capsulatum, and yeast-like H. capsulatum, were in the following ranges: 350-5720 µg/mL, 20-2860 µg/mL, and 40-1420 µg/mL, respectively for terpinen-4-ol; 250-4000 µg/mL, 30-2000 µg/mL, and 10-1000 µg/mL, respectively, for tyrosol; and 0.48-7.8 µg/mL, 0.25-16 µg/mL, and 0.125-4 µg/mL, respectively for β-lapachone. These compounds showed a decrease in MIC when the samples were subjected to osmotic stress, suggesting that the compounds acted on the fungal membrane. All the compounds were able to reduce the ergosterol content of the fungal strains. Finally, tyrosol was able to cause a leakage of intracellular molecules.

Simultaneous determination of five active compounds in wild and culture materials of Rhodiola crenulata by RP-HPLC / 中国药学杂志

OBJECTIVE: To establish a method for simultaneous determination of five components in wild and tissue-culture materials of Rhodiola crenulata. METHODS: The determination was performed on a Thermo-C18 column (4.6 mm×250 mm, 5 μm) with mobile phase composed of methanol-water (32:68) at a flow rate of 0.8 mL·min-1. The injection volume was 10 μL, column temperature was set at 30℃, and the detection wavelength was set at 277 nm at 0-13 min and 250 nm at 13-60 min. RESULTS: There were linear relationships between the peak areas and contents of salidroside, p-tyrosol, rosarin, rosavin and rosin in the ranges of 2.80-280.00 (r=0.9998), 2.80-280.00 (r=0.9997), 1.20-120.00 (r=0.9996), 1.60-160.00 (r=0.9997) and 1.20-120.00 μg·mL-1 (r=0.9997), respectively. The extraction recoveries varied from 99. 32% to 100.45%. CONCLUSION: The established method is simple and accurate for quality control of wild and tissue-culture materials of Rhodiola.

Simultaneous determination of eight constituents in Compound Rhodiola Capsule by HPLC-UV-ELSD / 中草药

Objective: To develop an HPLC-UV-ELSD method for the simultaneous determination of crenelatin, gallic acid, salidroside, tyrosol, harpagide, harpagoside, angoroside C, and cinnamic acid in Compound Rhodiola Capsule. Methods: Kromasil C18 column (250 mm×4.6 mm, 5 μm) was adopted. The mobile phase was composed of acetonitrile (A) and 0.3% HAC (B) with gradient elution. The flow rate was 1.0 mL/min and the detection wavelength was 275 nm. the column temperature was 30℃, and the evaporative light-scattering detector (ELSD) drift tube temperature was 40℃, and gas pressure of 1.5 bar (150 kPa). Results: Crenelatin, gallic acid, salidroside, tyrosol, harpagide, harpagoside, angoroside C, and cinnamic acid were separated well. The linear calibration curves were obtained in 67.084-670.84 μg/mL for crenelatin, R2=0.9992; 11.410-114.100 μg/mL for gallic acid, R2=0.9994; 78.995-789.95 μg/mL for salidroside, R2=0.9996; 19.625-196.25 μg/mL for tyrosol, R2=0.9997; 59.368-593.68 μg/mL for harpagide, R2=0.9998; 62.585-625.85 μg/mL for harpagoside, R2=0.9995; 55.045-550.45 μg/mL for angoroside C, R2=0.9996; and 6.895-68.95 μg/mL for cinnamic acid, R2=0.9998. The average recoveries of the eight constituents were 100.8%, 98.9%, 100.1%, 100.8%, 98.9%, 99.6%, 100.7%, and 99.2% with RSD of 0.64%, 0.56%, 0.35%, 0.65%, 0.26%, 0.58%, 1.00%, and 0.64%. Conclusion: The method is convenient, accurate, and can be used for the simultaneous determination of the preparation.

Influence of Tyrosol on Cell Growth Inhibition of KB Human Oral Cancer Cells

Tyrosol, a phenylethanoid and a derivative of phenethyl alcohol, possesses various biological properties, such as anti-oxidative and cardioprotective activity. Olive oil is the principal source of tyrosol in the human diet. However, so far the anti-cancer activity of tyrosol has not yet been well defined. This study therefore undertakes to examine the cytotoxic activity and the mechanism of cell death exhibited by tyrosol in KB human oral cancer cells. Treatment of KB cells with tyrosol induced the cell growth inhibition in a concentration- and a time-dependent manner. Furthermore, the treatment of tyrosol induced nuclear condensation and fragmentation of KB cells. Tyrosol also promoted proteolytic cleavage of procaspase-3, -7, -8 and -9, increasing the amounts of cleaved caspase-3, -7, -8 and -9. In addition, tyrosol increased the levels of cleaved PARP in KB cells. These results suggest that tyrosol induces the suppression of cell growth and cell apoptosis in KB human oral cancer cells, and is therefore a potential candidate for anti-cancer drug discovery.

Ultrasonic Extraction Process Research and Mathematic Simulation of Tyrosol in Rhodiolae Crenulatae Radix et Rhizoma / 中国中医药信息杂志

Objective To study the ultrasonic extraction technology of tyrosol in Rhodiolae Crenulatae Radix et Rhizoma;To mathematically simulate the extraction process.Methods The content of tyrosol was set as index;HPLC was used;the ultrasonic extraction process of tyrosol in Rhodiolae Crenulatae Radix et Rhizoma was optimized by an orhogonal experiment and mathematic simulation.Results The optimum ultrasonic extraction conditions were as follows:80%ethanol was used as extraction solvent, the ratio of material to liquid was 1:35 (g:mL), with the extraction time of 30 min and ultrasonic power of 240 W. In these conditions, the extracting rate of tyrosol was 0.150 1%. Compared with the heating reflux method, the extraction time should be shortened by 66.7% and the extracting rate should be increased by 12%.Conclusion The extraction method is simple and the extraction rate of effective components is high. Mathematical simulation values based on ultrasonic extraction are consistent with experiment values.

HPLC method for simultaneous quantitative determination of six active ingredients in Rhodiola crenulata from different origins of Qinghai-Tibet Plateau / 中草药

Objective: To analyze the six active ingredients in Rhodiola crenulata from the different origins of Qinghai-Tibet Plateau, to establish quantitative analysis methods, and to provide a basis for further use. Methods: Samples were extracted with 30% ethanol; In Diamonsil C18 column (250 mm × 4.6 mm,5 μm), gradient elution was carried out with acetonitrile -0.3% phosphoric acid solution as the mobile phase, detection wavelength 275 nm, and column temperature 25 ℃. Results: A good linear relationships of gallic acid, salidroside, tyrosol, catechin, ethyl gallate, and coumaric acid were at 38.2-382.0 (r = 0.9998), 301.0-3010.0 (r = 0.9999), 19.8 - 198.0 (r = 0.9996), 17.1-171.0 (r = 0.9996), 4.5-45.0 (r = 0.9998), and 6.38-63.80 ng/mL (r = 0.9994), respectively. Conclusion: The method is simple, rapid, accurate, and can be used simultaneously to determine the content of the six active ingredients in R. crenulata from the different origins of Qinghai-Tibet Plateau.

Study on the transcription profiling of biofilm formation by Candida albicans at different phases exposed to tyrosol and farnesol / 中华微生物学和免疫学杂志

Objective To investigate the effects of tyrosol and farnesol on the transcription profiling of C.albicans biofilm by microarray analysis.Methods The standard strain of C.albicans,SC5314 were cultured into four groups (tyrosol treated,farnesol treated,tyrosol and farnesol co-treated,and untreated control).The cell suspensions of SC5314 were prepared and dispensed into polystyrene flasks to form biofilm.Then,the biofilms were collected at 6 h and 24 h respectively after culturing.RNA samples were extracted and synthesized into cDNA through reverse transcription.The genome arrays were scanned with a confocal LuxScanTM scanner and the images were then analyzed by using LuxScanTM 3.0 software (both from CapitalBio).Bioinformatics analysis of the data was carried out by comparatively analyzing S.cerevisiae gene in KEGG gene database.Results The cDNA microarray data showed that tyrosol and farnesol regulated biofilm formation by regulating the genes associated with biofilm formation such as hypha genes and yeast genes.Tyrosol positively regulated gene expression of C.albicans biofilm,while farnesol played negative role.There were striking differences in gene expression patterns between tyrosol or farnesol treated groups and the control.Tyrosol had no antagonistic effect to farnesol.Bioinformatics analysis showed that the differential gene expression was involved in biological process,molecular function process and cell component formation.These genes regulated the C.albicans biofilm formation by encoding proteins involved in the biological metabolism.Conclusion Tyrosol and farnesol influenced the formation of C.albicans biofilm through regulating gene expression which showed differences at different phases of biofilm formation.

Chemical characterization of bioactive compounds from the endophytic fungus Diaporthe helianthi isolated from Luehea divaricata

Endophytic microorganisms, defined as fungi or bacteria that colonize the interior of plants without causing any immediate negative effects or damages, have reciprocal relationships with host plants. In some cases their presence is beneficial to the host due to the synthesis of bioactive compounds, among which several alcohols, esters, ketones and others that may react with other compounds and may be lethal to pathogenic microorganisms. Diaporthe helianthi (Phomopsis helianthi in its anamorphic phase) is available worldwide, especially in Europe, Asia and America. Isolated in Europe as an agent of the sunflower stem cancer, it has also been endophytically isolated from tropical and temperate plants. A D. helianthi strain isolated from Luehea divaricata has been employed in current research. An investigation of the secondary metabolite from D. helianthi by CC and NMR of ¹H and 13C yielded the separation of 10 fractions and the identification of the phenolic compound 2(-4 hydroxyphenyl)-ethanol (Tyrosol). Its antimicrobial reaction was tested and the ensuing antagonistic effects on the human pathogenic bacteria Enterococcus hirae, Escherichia coli, Micrococcus luteus, Salmonella typhi, Staphylococcus aureus, phytopathogenic Xanthomonas asc. phaseoli and phytopathogenic fungi were demonstrated. Results show that bioactive compounds and Tyrosol produced by D. helianthi have a biotechnological potential.

The role of cell density in the morphology of the Candida albicans bioflims development and the tyrosol production / 中华微生物学和免疫学杂志

Objective To study the role of cell density in the tyrosol production and morphology for Candida albicans biofilms. Methods C. albicans SC5314 and clinical isolates were propagated in yeast peptone dextrose (YPD) medium. Cells were collected by centrifugation and washed twice in sterile phosphate-buffered saline (PBS) before this study, then resuspended in RPMI 1640 supplemented with L-glutamine and adjusted to a desired concentration of 5 × 10~6 cells/ml, 5×10~5 cells/ml, 5 × 10~4 cells/ml, 5 × 10~3 cells/ml after counting with a hematocytometer. Standardized C. albicans cells were prepared as above description and 2000 μl of this standardized cell suspension was dispensed into the wells, then C. albicans biofilms were formed on the bottom of the polystyrene wells. In this study, tyrosol synthesized by SC5314 and clinical isolates of C. albicans biofilm was quantified by high performance liquid chromatography (HPLC). The effects of tyrosol on morphology of C. albicans biofilms were investigated by scanning electron microscopy (SEM). Results Tyrosol production of C. albicans biofilms was affected by cell densities. At lower inoculation size(5 μ 10~3 cells/ml), there was too less tyrosol production to be detected at the early stage of the biofilms formation. At higher inoculation size (5 μ10~6 cells/ml), tyrosol can be detectable at the early stage or at the mature stage of biofilms formation. There was a sharp increase in tyrosol concentration at 24 h, while there was a decrease in tyrosol concentration after that time from the strains when the strains were at an inoculation size of 5 × 10~6 cells/ml and 5 × 10~5 cells/ml. Cell densities affected the morphology formation of the C. albicans biofilms. At the early stage of the biofilms formation, C. albicans grew less germ tube at lower cell densities than that at the higher cell densities. With the mature of the biofilms, C. albicarts grew more hyphae at higher cell densities than that at the lower cell densities. All these above showed that cell densities played an important role in the propagation for the C. albi-cans in the biofilm formation. Conclusion Cell density play an important role in the formation of the C. albi-cans biofilms and the production of the tyrosol.

Preliminary study on the effect of tyrosol and farnesol on biofilm formation of Candida albicans / 中华微生物学和免疫学杂志

Objective To study the regulation of quorum sensing molecule tyrosol and farnesol on biofilm formation of Candida albicans. Methods Candida albicans biofilms of clinic isolates and standard strain SC5314 were built when quorum sensing molecule existed. And inverted microscope was used to observe the morphology of C. albicans cells. RT-PCR and MTT assay were carried out to investigate the effect of quorum sensing molecule on expression of the two genes (HTA1 and EFG1) and cytoactive. Results Tyrosol could not promote hyphae development and cytoactive of C. albicans biofilms. The expression of HTA1 of C. albicans in biofilms was up-regulated by tyrosol but EFG1 was not. The inhibitory effect of farnesol on hyphae development, cytoactive and gene expression were not changed by addition of tyrosol. Conclusion Tyrosol can make C. albicans biofilms active in early stage. But when tyrosol and farnesol were simultaneously added, the effect of tyrosol were masked by farnesol. And C. albicans cells were more sensitive to farnesol than to tyrosol.

Enhancement of NADPH: quinone oxidoreductase 1 (NQO_1) gene expression and inhibition of cell proliferation induced by ?-tyrosol in human hepatoma SMMC-7721 cells / 中国病理生理杂志

AIM: To investigate whether ?-tyrosol could induce expression of DT-diaphorase (NQO 1) gene and inhibit proliferation of hepatoma cells and their relationship.METHODS: The DT-diaphorase activity, mRNA expression and cell proliferation were examined using direct measurement of DT-diaphorase from cells cultured in microtiter wells, semi-quantitative reverse transcription-PCR (RT-PCR) technique and crystal violet assay after treatment of SMMC-7721 cells with ?-tyrosol for 24 h. RESULTS: Treatment of SMMC-7721 cells with ?-tyrosol caused an increase in DT-diaphorase activity and DT-diaphorase mRNA expression in a dose-dependent manner. A statistically significant correlation between DT-diaphorase mRNA levels and enzyme activities was evident. In almost the same concentration range of 70 mg/L to 100 mg/L, ?-tyrosol resulted in a dose-dependent inhibition of cell proliferation which well inversely correlated with DT-diaphorase activity.CONCLUSION: ?-tyrosol can induce DT-diaphorase activity and enhance expression of its gene, which may contribute to antiproliferation of hepatoma cells caused by the same chemical.

Effects of Fructus Ligustri Lucidi on the tyrosinase activity and melanogenesis of cultured human melanocytes / 中华皮肤科杂志

Objective To study the effects of tyrosol and oleanolic acid, two monomers of Fructus Ligustri Lucidi on the proliferation, tyrosinase activity and melanogenesis of cultured human melanocytes. Methods The cell proliferation was detected by MTT assay, the tyrosinase activity by dopa oxidase assay, the melanin content by NaOH - assay, and the mRNA level of tyrosinase ( TYR ) and tyrosinase related protein-1 (TRP-1) by RT-PCR. Results Incubation of tyrosol with cultured human melanocytes for 72 hours resulted in a significant stimulation of the tyrosinase activity and melanogenesis in a dose dependent manner. Compared with those in the cells without any treatment, the TYR and TRP-1 mRNA levels in the tyrosol -treated cells increased by 39.10% and 99.26%, respectively. Oleanolic acid also increased tyrosinase activity ( P

Determination of marker compounds in plants of Rhodiola L. from different habitats by RP-HPLC / 中草药

ObjectTo develop the reliable RP-HPLC methods for the determination of salidroside, tyrosol, rosavin, rosin, and rosarin in the plants of Rhodiola L. and to evaluate their species from different habitats. Methods Method Ⅰ: methanol-water (0.5 mmol/L SDS in 1% acetic acid aqueous solution) system for the analysis of salidroside; method Ⅱ: acetonitrile-water system for rosavin; method Ⅲ: aqueous acetonitrile-phosphoric gradient system for salidroside, tyrosol, rosavin, rosin, and rosarin. Results The contents of salidroside in different species range from 0.021% to 1.420%, and those of rosavin in all species are very limited or undetected except in Rhodiola rosea L. and R. sachalinensis. The contents of the five marker ingredients are significantly species- and habitat-dependent. Conclusion Three RP-HPLC methods are established for quantitative analysis of the above five marker ingredients in the meantime, respectively. Evaluation of the quality of varied species of Rhodiola L. shows that R. rosea growing in Xinjiang Uygur Autonomous Region and R. sachalinensis growing in Jilin province are the two better species contained with abundant above-mentioned ingredients in China.

Qualitative and quantitative analysis of salidroside and p-tyrosol in Hongjingtian Injection / 中成药

AIM:To establish the qualitative and quantitative analysis of salidroside and p-tyrosol in Hongjingtian Injection(Radix et Rhizoma Khodiolae crenulatae).METHODS:The identification of salidroside and p-tyrosol were performed by TLC.Two components were determined by HPLC.RESULTS:The TLC method was suitable for the identification of salidroside and p-tyrosol with good reproducibility.The average recoveries of salidroside and p-tyrosol were 97.6% and 99.1% with RSD of 1.10% and 2.69%(n=5),respectively.CONCLUSION:This convenient,sensitive,reliable method can be used as qualitative and quantitative analysis of salidroside and p-tyrosol in Hongjingtian Injection.