• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Simultaneous determination of four Sudan dyes in rat blood by UFLC-MS/MS and its application to a pharmacokinetic study in rats☆

    2015-11-17 01:23:52HoZhuYijunChenChngshunHungYngyngHnYiweiZhngShucnXieXiohongChenMicongJin
    Journal of Pharmaceutical Analysis 2015年4期
    關(guān)鍵詞:距離感面孔親人

    Ho Zhu,Yijun Chen,Chngshun Hung,*,Yngyng Hn,Yiwei Zhng,Shucn Xie,Xiohong Chen,Micong Jin

    aDepartment of Anesthesia,the First Hospital of Ningbo City,Ningbo 315010,China

    bZhejiang Provincial Key Laboratory of Health Risk Appraisal for Trace Toxic Chemicals,Ningbo Municipal Center for Disease Control and Prevention,Ningbo 315010,China

    cNingbo Key Laboratory of Poison Research and Control,Ningbo Municipal Center for Disease Control and Prevention,Ningbo 315010,China

    Original Article

    Simultaneous determination of four Sudan dyes in rat blood by UFLC-MS/MS and its application to a pharmacokinetic study in rats☆

    Hao Zhua,Yijun Chena,Changshun Huanga,*,Yangyang Hana,Yiwei Zhanga,Shucan Xiea,Xiaohong Chenb,c,Micong Jinb,c

    aDepartment of Anesthesia,the First Hospital of Ningbo City,Ningbo 315010,China

    bZhejiang Provincial Key Laboratory of Health Risk Appraisal for Trace Toxic Chemicals,Ningbo Municipal Center for Disease Control and Prevention,Ningbo 315010,China

    cNingbo Key Laboratory of Poison Research and Control,Ningbo Municipal Center for Disease Control and Prevention,Ningbo 315010,China

    A R T I C L E I N F O

    Article history:

    1 March 2015

    Accepted 2 March 2015

    Available online 18 March 2015

    Multiple reaction monitoring

    Pharmacokinetics

    Rat whole blood

    Sudan dye

    UFLC-MS/MS

    A rapid and sensitive method based on ultrafast liquid chromatography-tandem mass spectrometry was developed and validated for simultaneous determination of Sudan I,Sudan II,Sudan III,and Sudan IV levels in rat whole blood.Cleanert C18 mixed-mode polymeric sorbent was used for effective solid-phase extraction cleanup.Separation was carried out on a reversed-phase C18column(100 mm×2.1 mm,1.8 μm)using 0.1%(v/v)formic acid in water/0.1%(v/v)formic acid in acetonitrile as the mobile phase in gradient elution.Quantification was performed by an electrospray ionization source in the positive multiple reaction monitoring mode using D5-Sudan I as the internal standard.Calibration curves showed good linearity between 0.2 and 20.0 μg/L,with correlation coefficients higher than 0.9990.The average recovery rates were between 93.05%and 114.98%.The intra-and inter-day relative standard deviations were within 6.2%.The lower limit of quantification was 0.2 μg/L.All the analytes were found to be stable in a series of stability studies.The proposed method was successfully applied to a pharmacokinetic study of four Sudan dyes after oral administration to rats.

    ?2015 Xi'an Jiaotong University.Production and hosting by Elsevier B.V.All rights reserved.This is an open access article under the CC BY-NC-ND license(http://creativecommons.org/licenses/by-nc-nd/4.0/).

    1.Introduction

    Sudan dyes(Sudan I,Sudan II,Sudan III,and Sudan IV;Fig.1),a group of synthetic fat-soluble colorants that contain an azo group(-N=N-)as part of the structure,are mainly used in textile,rubber,plastic,paint,and other coloring applications.Sudan dyes are frequently used as a food-coloring agent by unscrupulous merchants because of their bright red color,colorfastness,and low price.However,these synthetic colorants have been classified as possible human carcinogens[1-4],and pose a potential risk to consumer health if their daily intake exceeds the maximally permitted levels established by the World Health Organization[5]. Thus,their extensive use in food products has been prohibited by the European Commission[6].

    Several analytical methods have been used to determine the presence of Sudan dyes in food products.These methods mainly involve liquid chromatography(LC)[7-11],flow injection chemiluminescence[12],enzyme-linked immunosorbent assays[13-15],capillary electrophoresis[16],electrochemical methods[17,18],LC-mass spectrometry(MS)[10,19-21],and electrospray tandem quadrupole orthogonal-acceleration time-of-flight MS[22-25].LC separation,most commonly used method,has been coupled with different spectrophotometric detection methods to determine the Sudan dye levels.These detections include MS[10,19-25],ultraviolet/visible detection[7,8,10,11],diode array detection[9],and electrochemical detection[12].However,relatively few studies have analyzed azo dye residues in whole blood samples[26].With the development of electrospray ionization(ESI),MS has become a powerful analytical tool in pharmaceutical analysis because of its high sensitivity,selectivity,and short run time[27-31].In combination with LC,tandem MS(MS/MS)is a superior alternative for drug residue analysis in biological samples.The aim of this study was to develop a sensitive,robust,and fast method for the analysis of low-level concentrations of four azo dyes(Sudan I,Sudan II,Sudan III,and Sudan IV)in whole blood samples using ultrafast LC with MS/MS(UFLC-MS/MS).The proposed method can achieve rapid separation of dyes in gradient elution within 6.5 min.The validated method was applied to determine the levels of the four Sudan analytes in rat whole blood samples obtained from healthy Sprague-Dawley(SD)rats used in pharmacokinetic studies.

    2.Experimental

    2.1.Chemicals and materials

    Sudan I(>95.0%),Sudan II(>99.0%),Sudan III(>98.0%),and Sudan IV(>98.0%)were purchased from Dr.Ehrenstorfer GmbH(Augsburg,Germany)and used without further purification.D5-Sudan I(purity>99.4%)was purchased from Witega Laboratories Berlin-Aldershof GmbH(Berlin,Germany).High-performance LC(HPLC)-grade acetonitrile and n-hexane were purchased from Merck(Darmstadt,Germany).HPLC-grade formic acid was obtained from Sigma-Aldrich(St.Louis,MO,USA).The solid-phase extraction(SPE)Cleanert C18 cartridges(500 mg/6 mL)used for sample preparation were procured from AgelaTechnologies(Tianjin,China).HPLC-grade water was generated using a Milli-Q integral water purification system (Millipore,Molsheim,F(xiàn)rance). Edible oil was obtained from the local market(Ningbo,China). Blank blood was obtained from untreated animals and frozen at-20°C until analysis.

    2.2.Animals

    Certified commercial SD rats,weighing 240±20 g,were purchased from the Experimental Animal Center of the Zhejiang Academy of Medical Sciences(Hangzhou,Zhejiang,China).Thirty healthy and drug-free SD rats(15 males and 15 females)were acclimatized in 10 rat cages in a unidirectional airflow room with controlled temperature(22-25°C),relative humidity(45%-55%),and a 12 h light/dark cycle for one week before the start of the experiments.The rats were given free access to filtered tap water and commercial rat food.

    2.3.Apparatus and conditions

    UFLC-MS/MS was performed using a Shimadzu Prominence UFLC XR system coupled with an Applied Biosystems SCIEX Triple Quad 5500 mass spectrometer and ESI source.The UFLC-MS/MS system was controlled,and data were analyzed on a computer equipped with Analyst 1.5.1(Applied Biosystems,MA,USA). Chromatographic separation was performed on an Agilent Eclipse Plus C18column(column size:2.1 mm×100 mm,particle size: 1.8 μm)using solution A(water with 0.1%(v/v)formic acid)and solution B(acetonitrile with 0.1%(v/v)formic acid)as the mobile phase.The flow rate was 0.45 mL/min.The gradient program used for elution was as follows:initial conditions with solvent B were increased from 10%B to 95%B in 2.0 min,held constant at 95%B for 2.5 min,returned to initial conditions,and maintained for 2.0 min for equilibration.The total run time was 6.5 min.The column temperature was set at 40°C.A total of 5 μL of the solution was injected into the UFLC-MS/MS system for analysis.

    MMSS wwaass ppeerrffoorrmmeedd uussiinngg tthhee ppoossiittiivvee EESSII mmooddee aanndd mmuullttiippllee reaction monitoring(MRM)mode for quantification.The optimized instrument operating parameters for mass spectral acquisition were as follows:ion spray voltage,5500 V;curtain gas,40 psi;interface heater,on;nebulizer gas(gas 1)and heater gas(gas 2),30 psi each;turbo spray temperature,500°C;entrance potential,10 V;and collision cell exit potential,10 V.Nitrogen was used in all cases.Retention times,precursor ions(Q1),product ions(Q3),declustering potential(DP),and collision energies(CEs)of each Sudan dye are shown in Table 1.The dwell time was set at 0.05 s for all the compounds.

    2.4.Standard stock solution preparation

    Individual stock solutions of four Sudan dyes and D5-Sudan I as an internal standard(IS)at 1000.0 mg/L were prepared in acetonitrile.The standard solution was stored at-20°C in tightly closed bottles in the dark.Mixed standard stock solution was prepared by diluting the standard stock solution of each dye with acetonitrile to a final concentration of 100.0 μg/L.The IS solution was subsequently diluted with acetonitrile to a final concentration of 100.0 μg/L.Working standard solutions at the concentrations of 0.2,0.5,1.0,2.0,5.0,10.0,and 20.0 μg/L were prepared daily by diluting the mixed standard stock solution with acetonitrile in appropriate proportions.

    2.5.Calibration standards and quality control(QC)samples

    Calibration standard solutions at the concentrations of 0.2,0.5,1.0,2.0,5.0,10.0,and 20.0 μg/L were freshly prepared by adding the appropriate mixed standard stock solution(100.0 μg/L)to 20.0 μL of blank rat whole blood,and stored at 4°C until the initiation of UFLC-MS/MS analysis(IS concentration:2.0 μg/L).The four concentration levels(0.2,0.5,5.0,and 16.0 μg/L)(lower limit of quantitation(LLOQ),low,medium,and high levels)in blank rat whole blood were considered as QC samples.These samples were stored in a freezer compartment at-20°C and brought to room temperature before use.

    2.6.Sample preparation

    Approximately 20.0 μL of whole blood sample and 10.0 μL of IS solution(100.0 μg/L)were mixed in a 2.0 mL Eppendorf tube,in which 0.5 mL of acetonitrile was added prior to extraction.The mixture was extracted by vortex-mixing for 3 min at ambient temperature.The extract was centrifuged at 20,000g for 5 min.The supernatant solution was then subjected to SPE for further purification.

    Fig.1.Chemical structures of Sudan I,Sudan II,Sudan III,and Sudan IV.

    The Cleanert C18 cartridges were preconditioned and equilibrated with 2 mL of methanol and 2 mL of water.Then,the aforementioned mixed solution was slowly loaded into thecolumn.Sampling was followed by a cleanup step.The cartridges were rinsed with 2 mL of acetonitrile/water(50:50,v/v)at a flow rate of 1 mL/min and pump-dried.Elution was performed with two aliquots of 3 mL of n-hexane.The eluate was evaporated to dryness by a gentle nitrogen flow and reconstituted with 0.5 mL of acetonitrile.Finally,the reconstituted eluate was filtered through a 0.2 μm polytetrafluoroethene(PTFE)microporous film,and 5.0 μL was injected into the UFLC-MS/MS system.

    2.7.Selectivity

    Six individual rat whole blood samples obtained from different sources were used to assess selectivity.Blood from each rat subject was extracted and checked for peaks that can interfere with the detection of the four Sudan dyes and IS in the rat whole blood samples.The chromatographic peaks were confirmed by comparing the retention times(tR)and mass fragment ions with those of reference standards.

    2.8.Matrix effect(ME)

    As suggested by Matuszewski et al.[26],a quantitative approach to assess absolute MEs was performed using a post-extraction addition study,in which percent ME(ME%)is calculated. In this study,the absolute ME was determined by comparing the mean peak areas of QC samples spiked post-extraction(B)with those of the standard solutions(A).ME%was determined using the following equation:absolute ME(ME%)=(B/A)×100.The absolute ME was evaluated at four concentration levels(LLOQ,0.2 μg/L;low,0.5 μg/L;medium,5.0 μg/L;and high,16.0 μg/L)and three parallels.

    2.9.Precision and accuracy

    The four QC samples(LLOQ,0.2 μg/L;low,0.5 μg/L;medium,5.0 μg/L;and high,16.0 μg/L)were used to assess interday precision and accuracy,which were analyzed on at least three separate runs.For intraday precision,six aliquots of each QC sample were thawed to room temperature and analyzed within a day.For interday precision,the tested experiments were done in triplicates for each QC sample on eight separate days within a two-week period.The relative standard deviations(RSD%)of the four Sudan dyes in each QC concentration were then calculated.

    Accuracy was expressed by the recovery rates,which were assessed by comparing the mean peak area ratios from the rat blood samples for standards spiked before extraction(C)with those for standards spiked after extraction into the plasma extracts(B).The recovery rates were calculated using the following formula:recovery(%)=(C/B)×100[26].Experiments on the three QC samples were performed in five replicates.

    2.10.Stability

    Stability experiments were performed to evaluate analyte stability in rat whole blood extracted samples under different conditions.The LLOQ,low,medium,and high-concentration QC samples were analyzed in triplicates.Short-term stability was tested by placing the QC samples at room temperature in an autosampler for 2,4,8,and 12 h.Freeze-thaw stability was determined by analyzing the samples for three cycles(from-20°C to room temperature).Long-term stability of the QC samples stored at-20°C was tested for 30 days.The stability experiments were then evaluated by comparing the concentration obtained with the standard values(the quantitative data of the freshly prepared QC samples by used the instrument method),followed by calculation of the deviations.

    2.11.Pharmacokinetic study

    The study was conducted in accordance with the Ethical Guidelines for Investigations in Laboratory Animals and approved by the Ethics Review Committee for Animal Experimentation of the Ningbo University.Certified commercial SD rats were used after growth for one week.Before the day of administration,30 rats were randomly assigned to five groups(blank edible oil group,Sudan I group,Sudan II group,Sudan III group,and Sudan IV group).Each group comprised three males and three females.Rats fasted overnight but were allowed access to water ad libitum.Sudan dyes(Sudan I,Sudan II,Sudan III,and Sudan IV)were separately dissolved in 10 mg/mL of edible oil and administered via a single oral(p.o.)dose of 50 mg/kg of Sudan dyes.Approximately 20.0 μL of the whole blood sample(at 10.0,20.0,30.0,and 40.0 min and 1.0,1.5,2.0,3.0,5.0,7.0,10.0,15.0,and 24.0 h postdosing for Sudan I and Sudan II;whereas 1.0,2.0,3.0,4.0,5.0,6.0,7.0,9.0,11.0,15.0,20.0,32.0,and 48.0 h postdosing for Sudan III and Sudan IV)was collected from the rat tail vein,poured into a 2 mL of heparinized centrifuge tube containing 0.5 mL of acetonitrile,and immediately vortex-mixed for 3 min.These samples were immediately centrifuged and cleaned according to Section 2.6.The extracts were stored in the dark at 4°C prior to analysis.

    Table 1Q1/Q3 ion pairs,DP,CE of MRM,and retention time for the optimized UFLC-MS/MS method in MRM mode.

    3.Results and discussion

    3.1.Sample preparation

    In this study,an efficient extraction solvent was selected using a recovery test(at 1.0 μg/L)in a rat whole blood sample.Based on their molecular structures and fat-soluble properties,various extraction solvents,such as methanol,acetonitrile,dichloromethane,and nhexane,were chosen.After the extraction and total processing time were comprehensively evaluated,a simple protein precipitation method was proposed for extraction.Acetonitrile was selected as the optimal protein precipitation solvent because of its good protein precipitation efficiency and good extraction efficiency.

    Given that Sudan dyes are fat-soluble compounds,normal neutral alumina SPE cartridges are recommended for use in cleanup.The activity of alumina N was adjusted according to the recoveries of standard solutions through the column because of variations in the quality of different batches of alumina N.During SPE cleanup,trace water in blood extracts significantly affected the recovery and precision.Thus,extracts should be dried with anhydrous sodium sulfate prior to sample loading.However,the application of this step for water-enriched rat whole blood samples prior to preparation is highly difficult.Therefore,we selected a stable and simple C18 SPE cartridge for cleanup,and achieved good recovery rates with all four dyes,in which>95%were unaffected by trace water.Fig.2 shows the MRM chromatograms with and without SPE cleanup.Interference significantly decreased with the use of SPE cleanup compared with that without SPE cleanup.

    Fig.2.MRM chromatograms with SPE cleanup(A)and without SPE cleanup(B).

    Fig.3.Representative MS/MS spectra obtained from Sudan I(A),Sudan II(B),Sudan III(C),and Sudan IV(D).

    3.2.LC and MS

    The analytes have hydrophobic and lipophilic characteristics. Thus,organic solvent-rich mobile phases are typically used for their rapid elution and strong retention in reversed-phase chromatography.C18columns from different manufacturers have been used in previous studies,in which the most common column length was 150 mm,but diameters and particles differed.In our preliminary experiments,mixtures of deionized water with two common HPLC organic modifiers(acetonitrile and methanol)were used as the mobile phases.The flow rate was 0.45 mL/min,and the sample injection volume was 5 μL.Acetonitrile offered better peaksymmetry and was selected for subsequent studies.The studied azo dyes were weak acids(pKa(SudanI&II)=11.65)because an intermolecular hydrogen bond could be formed with the phenolic hydroxyl groups.Thus,the effect of the mobile phase pH on the chromatographic behavior of analytes was investigated by acidifying the aqueous portion through the addition of different formicacid concentrations at 0%,0.05%,0.1%,0.2%,0.5%,and 1.0%(v/v). Significantly improved peak symmetry and signal response values were obtained at the formic acid concentration of≥0.1%.Thus,water with 0.1%(v/v)formic acid and acetonitrile with 0.1%(v/v)formic acid were finally selected as the mobile phase for gradient elution.

    Fig.4.Typical chromatograms of(A)blank rat blood;(B)blank rat blood spiked with Sudan I,Sudan II,Sudan III,and Sudan IV at LLOQ of 0.2 μg/L and IS of 2.0 μg/L;(C)rat blood sample obtained at 40 min and 5 h after a single oral administration of 50 mg/kg of Sudan I,Sudan II,Sudan III,Sudan IV,and IS(2.0 μg/L).

    Table 2Mean extraction recoveries and MEs of Sudan I,Sudan II,Sudan III,Sudan IV,and IS in SD rat whole blood samples(n=6).

    Table 3Intra-day and inter-day accuracy and precision values for the determination of the four analytes(n=3 days,six replicates per day).

    Table 4Regression equations,linear ranges,LODs,and LLOQs of the four analytes.

    In this study,the ESI-MS/MS behavior of four Sudan dyes was investigated in MRM mode.At a cone voltage of 25 V,the full-scan ESI(+)-MS spectra of the analytes showed only the protonated molecule[M+H]+,thereby confirming the molecular mass.Under ESI-MS/MS conditions,the product-ion mass spectra of Sudan dyes showed a characteristic fragmentation pattern for all the analytes,as shown in Fig.3.The protonated form of each analyte and IS,[M+H]+ion,was the parent ion in the Q1 spectrum and was used as the precursor ion to obtain the Q3 product ion spectra. The most sensitive mass transition was monitored at m/z 249.1→93.0 for Sudan I,m/z 277.1→121.2 for Sudan II,m/z 353.1→77.0 for Sudan III,m/z 381.2→91.0 for Sudan IV,and m/z 254.1→98.0 for IS.

    3.3.Photostability and“fast peaks”of Sudan III and Sudan IV

    高度的近視,讓我一生也沒有接觸過實(shí)實(shí)在在的東西。我只了解這個世界的一半。對親人、同學(xué)和同事,我永遠(yuǎn)看不清他們實(shí)在的面孔和真實(shí)的靈魂。對世界的看法,我總有一種模糊的距離感。如此霧里看花,讓我像一只迷途的羔羊。于是我成了一個自暴自棄的人。

    Previous studies reported the appearance of“fast-eluting”peaks in the chromatograms of Sudan III and Sudan IV[10,11,19]. Given that these peaks eluted a few minutes before the main peak of the compounds,they were called“fast peaks”,which were proven by M?lder et al.[10]to be caused by photochemical isomers via MS.This phenomenon could lead to about 10%-35% quantitative errors for Sudan III and Sudan IV if lightning conditions were not controlled.However,photo-induced isomerization was reversible when the compounds were stored in the dark or wrapped in aluminum foil for a sufficient time.

    A considerable increase in the peak area of isomers was observed upon standing on the tray of the autosampler without protection from light,which confirmed the findings of M?lder et al.(Fig.4).The nature of“fast peaks”could not be avoided in routine conditions.Thus,they were integrated together with the normal peaks for analysis.Our results(see Section 3.4)showed that the“fast peaks”did not affect the accuracy of the analysis when all the peaks of one compound were integrated together,and the RSD and accuracy were satisfactory.

    3.4.Method validation

    3.4.1.Selectivity

    Selectivity is defined as the ability of the bioanalytical method to measure a substance unequivocally and to discriminate between the analyte(s)and other components that may be present[32].Under optimized UFLC-MS/MS conditions,the rapid method yielded excellent selectivity and sensitivity for the analysis of Sudan I,Sudan II,Sudan III,Sudan IV,and IS in the blank whole blood samples(Fig.4).To demonstrate the selectivity of the method and to screen for interfering substances,three replicate analyses of six blank whole blood samples and blood spiked with the LLOQ were extracted and injected for analysis using the developed UFLC-MS/ MS method.The representative chromatograms of a blank rat blood sample and a blank rat blood sample spiked with four Sudan dyes(0.2 μg/L)are shown in Fig.4.No significant interfering endogenous peaks were observed at the retention times of the four Sudan dyes.Thus,according to the guidelines for industrial bioanalytical method validation(2001)[33],the method that we developed was selective.

    3.4.2.ME

    The details of the performed ME experiment are summarized in Table 2.The absolute MEs of the four compounds and IS ranged from 89.72%to 101.06%.An absent or insignificant ME in thedifferent sources was confirmed by the results,which agreed with the requirement of the guidelines for industry and bioanalytical method validation by the Food and Drug Administration[33].

    Table 5Stability assessment for Sudan I,Sudan II,Sudan III,and Sudan IV in SD rat whole blood samples(n=3).

    3.4.3.Precision and accuracy

    The accuracy and precision results are shown in Tables 2 and 3,respectively.The mean recovery rates for the four compounds(n=6)of the three concentrations ranged from 93.05%to 114.98%. The intra-day RSDs for 0.5,5.0,and 16.0 μg/L ranged from 1.6%to 6.2%,and their inter-day RSDs for 0.5,5.0,and 16.0 μg/L ranged from 1.3%to 4.8%.Thus,the repeatability and recovery of the assay were within the acceptance limits of±15%at the tested concentration levels.

    3.4.4.Linearity,limit of detection(LOD),and LLOQ

    The calibration model was selected based on the analysis of the data by linear regression with intercepts and 1/x2weighting factor by using the Least-Squares Refinement method.The linear calibration curves were plotted as the peak area ratio of the analyte to IS(Y)versus the target-compound mass-concentration ratios of the analyte to IS(X).Representative linear equations of the four compounds in rat whole blood are listed in Table 4.Each standard point in every calibration curve was back-calculated using its own equation.The linearities of the calibration curves were between 0.2 and 20.0 μg/L,and the correlation coefficients(r)of the four compounds were≥0.999.

    LODs were established based on signal-to-noise(S/N)ratio of 3,whereas LLOQs were established based on an S/N ratio of 10.The LODs for the four compounds in the injection solutions were estimated to be 0.06 μg/L.Based on the acceptable precision and accuracy,the LLOQ was 0.2 μg/L in the injection solutions,as shown in Table 4.

    3.4.5.Stability

    The results for freeze-thaw cycle stability and short-term stability under room temperature at 0.2,0.5,5.0,and 16.0 μg/L levels of the four compounds in rat whole blood are shown in Table 5.No significant degradations(<14.5%for short-term stability under room temperature,<13.5%for freeze-thaw cycle stability,and<8.5%for long-term stability)were observed for the four compounds.Results show that the four compounds were stable under the investigated conditions because the measured concentrations werewithintheacceptablelimits(≤15%ofthenominal concentrations).

    3.5.Pharmacokinetic study

    Fig.5.Mean blood concentration-time curves of Sudan I and Sudan II(A),and Sudan III and Sudan IV(B)in six SD rats after a single oral administration of 50 mg/kg of Sudan dyes.

    Table 6Pharmacokinetic parameters after oral administration of four Sudan dyes(50 mg/kg)to rats(n=6).

    The method that we developed was successfully applied to the pharmacokinetic study after a single oral administration of 50 mg/ kg Sudan I,Sudan II,Sudan III,and Sudan IV to SD rats.Mean blood concentration-time profiles of Sudan I,Sudan II,Sudan III,and Sudan IV are shown in Fig.5.The major pharmacokinetic parameters of the four analytes were calculated using a non-compartment model using DAS 2.0 statistical software(Pharmacology Institute of China),and the results are shown in Table 6.These pharmacokinetic results demonstrated that Sudan I,Sudan II,Sudan III,and Sudan IV reached the maximum point in the whole blood drug concentration of rats at about 0.6046,1.362,4.196,and 4.341 h after oral administration,respectively.The whole blood concentration decreased gradually with an elimination half-life(t1/2)of 0.8374,1.8102,2.3909,and 2.364 h for Sudan I,Sudan II,Sudan III,and Sudan IV,respectively.The absorption rate constants(ka)of Sudan I,Sudan II,Sudan III,and Sudan IV were 3.996,1.393,0.4368,and 0.4395 h-1,which were significantly greater than their elimination rate constants(ke)of 0.8277,0.3829,0.2899,and 0.2932 h-1,respectively.The four Sudan dyes were easily absorbed and quickly accessible to the loop body of the SD rats.The phenomenon of rapid absorption possibly involves the specific and lipophilic chemical properties of the four Sudan dyes.Given the double phospholipid compositions of the cell membrane of the rat,the lipophilic Sudan dyes successfully and rapidly accessed the organic body.These results were in accordance with the maximum blood concentrations achieved at 630.91,696.26,349.39,and 1304.61 μg/L for Sudan I,Sudan II,Sudan III,and Sudan IV,respectively.Theareasundertheconcentration-timecurve(AUC0-∞)of Sudan I,Sudan II,Sudan III,and Sudan IV were 1150.03,2966.66,2706.37,and10,013.91 μg h/L,respectively,which indicated the high bioavailability of the four analytes.The successful application of the UFLC-MS/MS method to the pharmacokinetic study of four Sudan dyes suggested its suitability and sufficiency for use in pharmacokinetic studies.

    4.Conclusions

    In this study,a sensitive and rapid UFLC-MS/MS method was developed and validated for determination of Sudan I,Sudan II,Sudan III,and Sudan IV levels in rat whole blood.The established method was fast,precise,accurate,specific,reproducible,and suitable for the pharmacokinetic study of Sudan I,Sudan II,Sudan III,and Sudan IV.The validated method was sufficiently sensitive with an LLOQ of 0.2 μg/L.Thus,the method successfully determined the Sudan I,Sudan II,Sudan III,and Sudan IV levels,which ranged from 0.2 μg/L to 20.0 μg/L.UFLC-MS/MS is suitable for detailed assessment in pharmacokinetic,bioequivalence,and bioavailability studies.

    Acknowledgments

    We would like to thank the Zhejiang Provincial Natural Science Foundation(No.LY13B050003),the Zhejiang Provincial Analytical Foundation of China(No.2013C37089),the Zhejiang Province Medical Health Foundation,China(No.2013KYA187),the Ningbo Natural Science Foundation in Zhejiang(No.2011A610058),the Advanced Key Program of Agriculture and Social Development Funds of Ningbo,China(No.2011C11021),and the Zhejiang Provincial Program for the Cultivation of High-level Innovative Health Talents(No.2011H1132)for financial support of this research.

    Appendix A.Supporting information

    Supplementary data associated with this article can be found in the online version at http://dx.doi.org/10.1016/j.jpha.2015.03.001.

    References

    [1]M.Stiborová,V.Martínek,H.Rydlová,et al.,Sudan I is a potential carcinogen for humans evidence for its metabolic activation and detoxication by human recombinant cytochrome P450 1A1 and liver microsomes,Cancer Res.62(2002)5678-5684.

    [2]M.Coulet,J.C.Leblanc,R.Woodrow Setzer,Application of the margin of exposure(MoE)approach to substances in food that are genotoxic and carcinogenic:EXAMPLE 12:Sudan I(CAS No.842-07-9),F(xiàn)ood Chem.Toxicol.48(2010)S106-S111.

    [3]K.K.Mishra,S.Dixit,S.K.Purshottam,et al.,Exposure assessment to Sudan dyes through consumption of artificially coloured chilli powders in India,Int.J. Food Sci.Technol.42(2007)1363-1366.

    [4]L.H.Ahlstrom,C.S.Eskilsson,E.Bjorklund,Determination of banned azo dyes in consumer goods,Trends Anal.Chem.24(2005)49-56.

    [5]A.Downham,P.Collins,Colouring our foods in the last and next millennium,Int.J.Food Sci.Technol.35(2000)5-22.

    [6]Commission Decision of 23 May 2005 on Emergency Measures Regarding Chilli Products,Curcuma and Palm Oil,(2005/402/EC)L135/34 Official Journal of the European Union.

    [7]Y.Fan,M.Chen,C.Shentu,et al.,Ionic liquids extraction of Para Red and Sudan dyes from chilli powder,chilli oil and food additive combined with high performance liquid chromatography,Anal.Chim.Acta 650(2009)65-69.

    [8]R.Noguerol-Cal,J.M.López-Vilari?o,G.Fernández-Martínez,et al.,High-performance liquid chromatography analysis of ten dyes for control of safety of commercial articles,J.Chromatogr.A 1179(2008)152-160.

    [9]P.Qi,T.Zeng,Z.Wen,et al.,Interference-free simultaneous determination of Sudan dyes in chili foods using solid phase extraction coupled with HPLCDAD,F(xiàn)ood Chem.125(2011)1462-1467.

    [10]K.M?lder,A.Künnapas,K.Herodes,et al.,“Fast peaks”in chromatograms of Sudan dyes,J.Chromatogr.A 1160(2007)227-234.

    [11]C.K.Zacharis,F(xiàn).S.Kika,P.D.Tzanavaras,et al.,Development and validation of a rapid HPLC method for the determination of five banned fat-soluble colorants in spices using a narrow-bore monolithic column,Talanta 84(2011)480-486.

    [12]Y.Zhang,Z.Zhang,Y.Sun,Development and optimization of an analytical method for the determination of Sudan dyes in hot chilli pepper by highperformance liquid chromatography with on-line electrogenerated BrO-luminol chemiluminescence detection,J.Chromatogr.A 1129(2006)34-40.

    [13]Y.H.Qi,W.C.Shan,Y.Z.Liu,et al.,Production of the polyclonal antibody against Sudan 3 and immunoassay of Sudan dyes in food samples,J.Agric.Food Chem. 60(2012)2116-2122.

    [14]Y.Wang,D.Wei,H.Yang,et al.,Development of a highly sensitive and specific monoclonal antibody-based enzyme-linked immunosorbent assay(ELISA)for detection of Sudan I in food samples,Talanta 77(2009)1783-1789.

    [15]D.Han,M.Yu,D.Knopp,et al.,Development of a highly sensitive and specific enzyme-linked immunosorbent assay for detection of Sudan I in food samples,J.Agric.Food Chem.55(2007)6424-6430.

    [16]E.Mejia,Y.Ding,M.F.Mora,et al.,Determination of banned Sudan dyes in chili powder by capillary electrophoresis,F(xiàn)ood Chem.102(2007)1027-1033.

    [17]M.J.Du,X.G.Han,Z.H.Zhou,et al.,Determination of Sudan I in hot chili powder by using an activated glassy carbon electrode,F(xiàn)ood Chem.105(2007)883-888.

    [18]L.Ming,X.Xi,T.Chen,et al.,Electrochemical determination of trace Sudan I contamination in chili powder at carbon nanotube modified electrodes,Sensors 8(2008)1890-1900.

    [19]C.Schummer,J.Sassel,P.Bonenberger,et al.,Low-level detections of Sudan I,II,III and IV in spices and chili-containing foodstuffs using UPLC-ESI-MS/MS,J. Agric.Food Chem.61(2013)2284-2289.

    [20]F.Calbiani,M.Careri,L.Elviri,et al.,Development and in-house validation of a liquid chromatography-electrospray-tandem mass spectrometry method for the simultaneous determination of Sudan I,Sudan II,Sudan III and Sudan IV in hot chilli products,J.Chromatogr.A 1042(2004)123-130.

    [21]F.Mazzotti,L.Di Donna,L.Maiuolo,et al.,Assay of the set of all Sudan azodye(I,II,III,IV,and Para-Red)contaminating agents by liquid chromatography-tandem mass spectrometry and isotope dilution methodology,J. Agric.Food Chem.56(2007)63-67.

    [22]C.Ferrer,A.R.Fernández-Alba,I.Ferrer,Analysis of illegal dyes in food by LC/ TOF-MS,Int.J.Environ.Anal.Chem.87(2007)999-1012.

    [23]J.Lau,C.K.Meng,J.Gushue,et al.,Analyzing Compounds of Environmental Interest Using an LC/Q-TOF—Part 1:Dyes and Pigments,Agilent Technologies Publication,5989-9336EN,August 2008.

    [24]F.Calbiani,M.Careri,L.Elviri,et al.,Accurate mass measurements for the confirmation of Sudan azo-dyes in hot chilli products by capillary liquid chromatography-electrospray tandem quadrupole orthogonal-acceleration time of flight mass spectrometry,J.Chromatogr.A 1058(2004)127-135.

    [25]Y.Fang,M.Zumwalt,Using TOF for Screening and Quantitation of Sudan Red Colorants in Food,Agilent application,Available from:〈http://www.agilent. com/〉.

    [26]B.K.Matuszewski,M.L.Constanzer,C.M.Chavez-Eng,Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS,Anal.Chem.75(2003)3019-3030.

    [27]A.Lévèques,L.Actis-Goretta,M.J.Rein,et al.,UPLC-MS/MS quantification of total hesperetin and hesperetin enantiomers in biological matrices,J.Pharm. Biomed.Anal.57(2012)1-6.

    [28]G.Lüthi,V.Blangy,C.B.Eap,et al.,Buprenorphine and norbuprenorphine quantification in human plasma by simple protein precipitation and ultra-high performance liquid chromatography tandem mass spectrometry,J.Pharm. Biomed.Anal.77(2013)1-8.

    [29]Y.Ling,Z.Li,M.Chen,et al.,Analysis and detection of the chemical constituents of Radix Polygalae and their metabolites in rats after oral administration by ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry,J. Pharm.Biomed.Anal.85(2013)1-13.

    [30]E.Marchei,E.Papaseit,O.Q.Garcia-Algar,et al.,Determination of atomoxetine and its metabolites in conventional and non-conventional biological matricesby liquid chromatography-tandem mass spectrometry,J.Pharm.Biomed. Anal.60(2012)26-31.

    [31]H.Wu,J.Guo,S.Chen,et al.,Recent developments in qualitative and quantitative analysis of phytochemical constituents and their metabolites using liquid chromatography-mass spectrometry,J.Pharm.Biomed.Anal.72(2013)267-291.

    [32]F.T.Peters,H.H.Maurer,Bioanalytical method validation and its implications for forensic and clinical toxicology-a review,Accredit.Qual.Assur.7(2002)441-449.

    [33]U.S.Department of Health and Human Services Food and Drug Administration,Center for Food Safety and Applied Nutrition,Guidance for Industry Bioanalytical Method Validation,2001.

    17 April 2014

    in revised form

    http://dx.doi.org/10.1016/j.jpha.2015.03.001

    2095-1779/?2015 Xi'an Jiaotong University.Production and hosting by Elsevier B.V.All rights reserved.This is an open access article under the CC BY-NC-ND license

    (http://creativecommons.org/licenses/by-nc-nd/4.0/).

    ☆Peer review under responsibility of Xi'an Jiaotong University.

    .Tel.:+86 574 87086437;fax:+86 574 87085078. E-mail address:nbhcs1967@163.com(C.Huang).

    猜你喜歡
    距離感面孔親人
    本期面孔
    遼河(2022年4期)2022-06-09 01:56:03
    你是我的親人
    草原歌聲(2021年2期)2021-10-14 08:27:42
    親人
    多變的面孔
    自然面孔
    人與自然(2019年4期)2019-07-26 12:52:37
    親人(連載二)
    寫自己的親人
    格斗距離感漫談
    少林與太極(2018年9期)2018-11-25 17:42:39
    江國香織文學(xué)中女性的孤獨(dú)
    北極光(2016年6期)2016-08-17 21:42:12
    格斗中距離感之管見
    亚洲精品视频女| 成年美女黄网站色视频大全免费| 久久精品国产亚洲av天美| 国产精品欧美亚洲77777| 老熟女久久久| 久久这里有精品视频免费| 如何舔出高潮| 亚洲精品一区蜜桃| 女人被躁到高潮嗷嗷叫费观| 国产亚洲欧美精品永久| 又粗又硬又长又爽又黄的视频| xxxhd国产人妻xxx| 久久精品国产综合久久久| 国产黄色视频一区二区在线观看| 久久青草综合色| 亚洲精品久久午夜乱码| 夫妻午夜视频| 久久国产亚洲av麻豆专区| 欧美少妇被猛烈插入视频| 两个人看的免费小视频| 我要看黄色一级片免费的| 久久人人97超碰香蕉20202| 欧美日韩亚洲高清精品| 91成人精品电影| 亚洲av综合色区一区| 久久国产精品男人的天堂亚洲| 久久亚洲国产成人精品v| 久久久久久久亚洲中文字幕| 亚洲国产成人一精品久久久| 久久久精品94久久精品| 丰满乱子伦码专区| 中国三级夫妇交换| 99久久中文字幕三级久久日本| 一本—道久久a久久精品蜜桃钙片| 国产男女内射视频| 91精品国产国语对白视频| 王馨瑶露胸无遮挡在线观看| 黄色视频在线播放观看不卡| 一级a爱视频在线免费观看| 国语对白做爰xxxⅹ性视频网站| 亚洲精品,欧美精品| 亚洲伊人久久精品综合| 日本vs欧美在线观看视频| 日韩三级伦理在线观看| 一二三四中文在线观看免费高清| 国语对白做爰xxxⅹ性视频网站| 在线免费观看不下载黄p国产| 老司机亚洲免费影院| 久久久久久久久免费视频了| 国产av国产精品国产| 久久久久久久久免费视频了| 天堂8中文在线网| 精品国产国语对白av| 黄网站色视频无遮挡免费观看| 日本爱情动作片www.在线观看| 精品少妇黑人巨大在线播放| 亚洲精品一二三| 曰老女人黄片| 一级毛片电影观看| 亚洲欧洲日产国产| 精品人妻偷拍中文字幕| 九草在线视频观看| 女人高潮潮喷娇喘18禁视频| 99九九在线精品视频| 精品卡一卡二卡四卡免费| 午夜福利影视在线免费观看| 欧美日韩av久久| 欧美亚洲日本最大视频资源| 精品人妻一区二区三区麻豆| 久热这里只有精品99| 人妻一区二区av| 免费高清在线观看日韩| 成人午夜精彩视频在线观看| 精品亚洲成国产av| 欧美日韩亚洲高清精品| 自线自在国产av| 涩涩av久久男人的天堂| 精品亚洲成国产av| 亚洲国产看品久久| 少妇被粗大的猛进出69影院| 亚洲美女搞黄在线观看| 久久狼人影院| 伦理电影大哥的女人| 一级片免费观看大全| 永久免费av网站大全| 国产国语露脸激情在线看| 久久久精品94久久精品| 欧美人与性动交α欧美软件| 久久精品久久久久久久性| 欧美97在线视频| 五月开心婷婷网| 男的添女的下面高潮视频| 高清黄色对白视频在线免费看| 久久国产精品大桥未久av| a级毛片在线看网站| 国产精品偷伦视频观看了| 成人手机av| 视频区图区小说| 国产精品成人在线| 大陆偷拍与自拍| 国产毛片在线视频| 欧美97在线视频| 精品国产一区二区三区四区第35| 久久人人爽av亚洲精品天堂| 久久人人爽av亚洲精品天堂| 国产一区二区 视频在线| 精品第一国产精品| 大码成人一级视频| 欧美日韩视频精品一区| 交换朋友夫妻互换小说| 美女脱内裤让男人舔精品视频| 两性夫妻黄色片| 国产一区二区在线观看av| 美女国产视频在线观看| 中文精品一卡2卡3卡4更新| 国产精品无大码| 国产成人精品久久二区二区91 | 亚洲欧洲国产日韩| 国产一区二区在线观看av| 国产精品三级大全| 啦啦啦在线观看免费高清www| 免费av中文字幕在线| 久久人人爽av亚洲精品天堂| 国产1区2区3区精品| 中文字幕精品免费在线观看视频| 精品国产一区二区三区久久久樱花| 一级a爱视频在线免费观看| 日日爽夜夜爽网站| 啦啦啦在线免费观看视频4| av国产久精品久网站免费入址| 午夜日韩欧美国产| videossex国产| 黄色视频在线播放观看不卡| 国产成人欧美| 亚洲第一av免费看| 丝袜喷水一区| 久久亚洲国产成人精品v| 国产一区二区激情短视频 | 成人午夜精彩视频在线观看| 高清视频免费观看一区二区| 久久精品人人爽人人爽视色| 亚洲欧美精品自产自拍| 2022亚洲国产成人精品| 久久精品国产亚洲av涩爱| 国产淫语在线视频| 欧美精品高潮呻吟av久久| 久久精品夜色国产| 91精品伊人久久大香线蕉| 国产成人a∨麻豆精品| 中国三级夫妇交换| 中文字幕最新亚洲高清| 夫妻午夜视频| www.熟女人妻精品国产| 青青草视频在线视频观看| 免费观看无遮挡的男女| 日本欧美国产在线视频| 日韩一区二区三区影片| 中文字幕亚洲精品专区| 国产av一区二区精品久久| 1024香蕉在线观看| 亚洲视频免费观看视频| 精品酒店卫生间| 日韩熟女老妇一区二区性免费视频| 99精国产麻豆久久婷婷| 人人妻人人爽人人添夜夜欢视频| 中文字幕制服av| 国产精品蜜桃在线观看| 蜜桃国产av成人99| 精品国产超薄肉色丝袜足j| 亚洲精品在线美女| 久久精品久久久久久久性| 国产精品蜜桃在线观看| 天天躁狠狠躁夜夜躁狠狠躁| 亚洲三级黄色毛片| 少妇被粗大的猛进出69影院| 欧美激情极品国产一区二区三区| 在线天堂中文资源库| av.在线天堂| 亚洲一码二码三码区别大吗| 人成视频在线观看免费观看| 超色免费av| 精品久久久久久电影网| 久久久久视频综合| 美女xxoo啪啪120秒动态图| 亚洲成人手机| 日韩 亚洲 欧美在线| 性色av一级| 国产精品久久久久久精品古装| 精品国产超薄肉色丝袜足j| 中文字幕亚洲精品专区| 欧美在线黄色| 看十八女毛片水多多多| 国产熟女欧美一区二区| 成年美女黄网站色视频大全免费| 日本av免费视频播放| av免费在线看不卡| 精品第一国产精品| 一本大道久久a久久精品| 三级国产精品片| 在线观看人妻少妇| 少妇被粗大的猛进出69影院| 高清视频免费观看一区二区| 亚洲一码二码三码区别大吗| 亚洲精品,欧美精品| 亚洲成国产人片在线观看| 国产熟女午夜一区二区三区| 两性夫妻黄色片| 男女免费视频国产| 国产福利在线免费观看视频| 亚洲成人av在线免费| 日韩,欧美,国产一区二区三区| 国产精品一区二区在线观看99| 80岁老熟妇乱子伦牲交| 人人妻人人澡人人爽人人夜夜| 亚洲中文av在线| 欧美xxⅹ黑人| 精品久久久久久电影网| 国产精品一区二区在线不卡| 大片免费播放器 马上看| 一级毛片我不卡| 侵犯人妻中文字幕一二三四区| av在线app专区| 日韩,欧美,国产一区二区三区| 久久久久久久久久久久大奶| 最近最新中文字幕免费大全7| 亚洲综合色网址| 在线观看三级黄色| av.在线天堂| 久久久久久久亚洲中文字幕| 国产日韩欧美在线精品| 两个人免费观看高清视频| 午夜福利,免费看| 久久国产精品大桥未久av| 日本-黄色视频高清免费观看| 国产黄频视频在线观看| 国产精品亚洲av一区麻豆 | 男的添女的下面高潮视频| 99热国产这里只有精品6| 国产精品免费大片| av国产久精品久网站免费入址| 国产精品99久久99久久久不卡 | 一级a爱视频在线免费观看| www.自偷自拍.com| 人人妻人人爽人人添夜夜欢视频| 交换朋友夫妻互换小说| 精品一区二区免费观看| 亚洲精品久久午夜乱码| 亚洲,一卡二卡三卡| 国产免费一区二区三区四区乱码| 人妻少妇偷人精品九色| 亚洲少妇的诱惑av| 黄色配什么色好看| 国产极品天堂在线| xxx大片免费视频| 日韩成人av中文字幕在线观看| 久久精品夜色国产| 99久久人妻综合| 久久av网站| 欧美精品人与动牲交sv欧美| 亚洲精品乱久久久久久| 久久韩国三级中文字幕| 国产精品久久久久久久久免| 久久久欧美国产精品| 男女免费视频国产| 一级毛片 在线播放| 卡戴珊不雅视频在线播放| 人妻人人澡人人爽人人| 午夜日韩欧美国产| 中文字幕精品免费在线观看视频| 国产亚洲一区二区精品| 日韩一区二区三区影片| 国产成人午夜福利电影在线观看| 国产成人午夜福利电影在线观看| 国产av码专区亚洲av| 伦理电影大哥的女人| 欧美日韩视频精品一区| 男女边吃奶边做爰视频| 免费日韩欧美在线观看| 精品人妻一区二区三区麻豆| 美女高潮到喷水免费观看| 黄片无遮挡物在线观看| 亚洲av电影在线观看一区二区三区| 曰老女人黄片| 亚洲第一av免费看| 中文字幕另类日韩欧美亚洲嫩草| 女人精品久久久久毛片| 91精品伊人久久大香线蕉| 啦啦啦啦在线视频资源| 国产深夜福利视频在线观看| 亚洲欧美成人精品一区二区| 最近最新中文字幕大全免费视频 | 色吧在线观看| 综合色丁香网| 下体分泌物呈黄色| 中文字幕制服av| 久久久欧美国产精品| 在线天堂最新版资源| 欧美人与性动交α欧美精品济南到 | 亚洲,欧美,日韩| 母亲3免费完整高清在线观看 | 亚洲情色 制服丝袜| 高清欧美精品videossex| 久久久久久久大尺度免费视频| 十八禁网站网址无遮挡| 久久久亚洲精品成人影院| 久久久久久伊人网av| 久久国内精品自在自线图片| 亚洲精品国产av成人精品| 亚洲精品国产av蜜桃| 成年动漫av网址| 麻豆av在线久日| 国产精品一区二区在线不卡| 久久久久国产精品人妻一区二区| 亚洲人成网站在线观看播放| 亚洲精品,欧美精品| 亚洲欧洲精品一区二区精品久久久 | 80岁老熟妇乱子伦牲交| 亚洲国产精品一区二区三区在线| 少妇的丰满在线观看| 91精品国产国语对白视频| 国产精品久久久久久精品古装| 午夜福利,免费看| 免费高清在线观看视频在线观看| av一本久久久久| 日韩中文字幕欧美一区二区 | 亚洲av成人精品一二三区| 美女福利国产在线| www.自偷自拍.com| 成人亚洲欧美一区二区av| 亚洲美女视频黄频| 男男h啪啪无遮挡| 99久久人妻综合| 黑丝袜美女国产一区| 国产成人91sexporn| 国产深夜福利视频在线观看| 18禁国产床啪视频网站| 另类亚洲欧美激情| 永久免费av网站大全| 婷婷色综合大香蕉| 亚洲国产精品一区三区| 一区二区三区乱码不卡18| 精品99又大又爽又粗少妇毛片| 国产精品久久久av美女十八| 满18在线观看网站| 亚洲一区二区三区欧美精品| 日韩三级伦理在线观看| 夜夜骑夜夜射夜夜干| 老汉色∧v一级毛片| 久久久a久久爽久久v久久| 免费高清在线观看日韩| 男女下面插进去视频免费观看| 在线观看三级黄色| 亚洲五月色婷婷综合| 亚洲国产精品999| 汤姆久久久久久久影院中文字幕| 一本—道久久a久久精品蜜桃钙片| 亚洲av免费高清在线观看| 老汉色∧v一级毛片| 91精品三级在线观看| 观看av在线不卡| 中国国产av一级| 丝袜脚勾引网站| 两个人免费观看高清视频| 成人毛片60女人毛片免费| 亚洲国产欧美在线一区| 高清欧美精品videossex| videos熟女内射| 大陆偷拍与自拍| 国产成人91sexporn| 精品少妇内射三级| 日韩制服丝袜自拍偷拍| 大香蕉久久网| 日韩精品有码人妻一区| 啦啦啦在线免费观看视频4| 久久久久久久国产电影| 精品人妻熟女毛片av久久网站| 国产 一区精品| 欧美精品av麻豆av| 丰满饥渴人妻一区二区三| 赤兔流量卡办理| 香蕉国产在线看| 在线观看免费高清a一片| 亚洲国产毛片av蜜桃av| 国产成人精品无人区| 免费久久久久久久精品成人欧美视频| 青春草国产在线视频| 成人午夜精彩视频在线观看| 男人操女人黄网站| 日韩欧美精品免费久久| 成年动漫av网址| 久久99精品国语久久久| 毛片一级片免费看久久久久| 亚洲欧美色中文字幕在线| 18在线观看网站| 精品少妇久久久久久888优播| 婷婷成人精品国产| 十八禁网站网址无遮挡| 久久这里只有精品19| 男人爽女人下面视频在线观看| 国产午夜精品一二区理论片| 久久99蜜桃精品久久| av在线播放精品| 久久精品国产亚洲av天美| 免费观看av网站的网址| 亚洲欧洲日产国产| 久热久热在线精品观看| 最近中文字幕2019免费版| 亚洲精品国产一区二区精华液| 另类精品久久| 性高湖久久久久久久久免费观看| 久久久久视频综合| 国产日韩欧美视频二区| 岛国毛片在线播放| 亚洲精品乱久久久久久| 国产乱来视频区| 精品国产一区二区三区久久久樱花| 夫妻午夜视频| 曰老女人黄片| 久久久欧美国产精品| 亚洲精品日韩在线中文字幕| 97在线视频观看| 丝袜脚勾引网站| 久久婷婷青草| 丝袜人妻中文字幕| www.熟女人妻精品国产| 丝瓜视频免费看黄片| 国产精品.久久久| 最近最新中文字幕免费大全7| 久久精品夜色国产| 人妻一区二区av| 桃花免费在线播放| 国产日韩欧美视频二区| 黑丝袜美女国产一区| 国产精品女同一区二区软件| 韩国精品一区二区三区| 高清黄色对白视频在线免费看| 精品少妇久久久久久888优播| 亚洲国产精品国产精品| 日产精品乱码卡一卡2卡三| 午夜老司机福利剧场| 久热这里只有精品99| 欧美bdsm另类| 亚洲一区中文字幕在线| 尾随美女入室| 极品人妻少妇av视频| 国产精品国产三级国产专区5o| 制服丝袜香蕉在线| 大片免费播放器 马上看| 日韩视频在线欧美| 波多野结衣一区麻豆| 在现免费观看毛片| 日韩在线高清观看一区二区三区| 欧美日韩一级在线毛片| 在线观看免费高清a一片| tube8黄色片| 亚洲人成电影观看| 欧美黄色片欧美黄色片| 国产精品 国内视频| 一级毛片电影观看| 国产一级毛片在线| 夫妻性生交免费视频一级片| 男女高潮啪啪啪动态图| 伊人久久国产一区二区| 只有这里有精品99| 欧美人与性动交α欧美软件| 亚洲,欧美,日韩| 国产男人的电影天堂91| 国产深夜福利视频在线观看| 亚洲国产精品一区二区三区在线| 精品少妇久久久久久888优播| 十八禁高潮呻吟视频| 欧美日韩综合久久久久久| 欧美另类一区| 欧美成人精品欧美一级黄| 成人毛片60女人毛片免费| 免费观看性生交大片5| 国产亚洲av片在线观看秒播厂| 日产精品乱码卡一卡2卡三| 免费观看性生交大片5| 欧美97在线视频| 亚洲av成人精品一二三区| 亚洲欧美一区二区三区国产| 久久久国产精品麻豆| 亚洲内射少妇av| 人妻系列 视频| 国产熟女欧美一区二区| 日本av手机在线免费观看| 国产xxxxx性猛交| 日韩精品有码人妻一区| 午夜激情av网站| 黑丝袜美女国产一区| 久久影院123| 久久狼人影院| 黄片无遮挡物在线观看| 久久国产亚洲av麻豆专区| 欧美+日韩+精品| 最新中文字幕久久久久| 欧美日韩国产mv在线观看视频| 日韩欧美一区视频在线观看| 亚洲精品,欧美精品| 亚洲国产色片| 国产毛片在线视频| 亚洲国产av新网站| 久久人人爽人人片av| 黄色配什么色好看| 丰满乱子伦码专区| 男女午夜视频在线观看| 在线观看免费日韩欧美大片| 美女国产高潮福利片在线看| 午夜福利在线免费观看网站| 黄片播放在线免费| 美女午夜性视频免费| 亚洲美女视频黄频| 韩国av在线不卡| 男女免费视频国产| 卡戴珊不雅视频在线播放| 日韩在线高清观看一区二区三区| 自拍欧美九色日韩亚洲蝌蚪91| 国产爽快片一区二区三区| 香蕉国产在线看| 中文乱码字字幕精品一区二区三区| 午夜福利在线免费观看网站| 观看美女的网站| 亚洲精华国产精华液的使用体验| 一本—道久久a久久精品蜜桃钙片| 蜜桃国产av成人99| 亚洲av日韩在线播放| 丝袜美足系列| 国产日韩欧美在线精品| 欧美日韩一级在线毛片| 国产乱来视频区| 男女免费视频国产| 久久精品国产综合久久久| 一本一本久久a久久精品综合妖精 国产伦在线观看视频一区 | 观看av在线不卡| 97精品久久久久久久久久精品| 天堂中文最新版在线下载| 久久人人爽av亚洲精品天堂| 精品人妻在线不人妻| 最近最新中文字幕免费大全7| 亚洲美女搞黄在线观看| 香蕉丝袜av| 精品少妇内射三级| 岛国毛片在线播放| 日韩av免费高清视频| 丁香六月天网| 日本wwww免费看| 欧美亚洲日本最大视频资源| 欧美xxⅹ黑人| 日韩免费高清中文字幕av| 女人久久www免费人成看片| 亚洲精品久久午夜乱码| 亚洲欧美成人精品一区二区| 精品一区在线观看国产| 一区福利在线观看| av又黄又爽大尺度在线免费看| www.自偷自拍.com| 免费观看a级毛片全部| 丰满饥渴人妻一区二区三| 亚洲av欧美aⅴ国产| 五月天丁香电影| 国产有黄有色有爽视频| 99久久综合免费| 成人亚洲精品一区在线观看| 久久久久人妻精品一区果冻| 午夜福利一区二区在线看| 国产成人精品婷婷| 一个人免费看片子| 日韩一卡2卡3卡4卡2021年| 国产深夜福利视频在线观看| 26uuu在线亚洲综合色| 另类精品久久| 麻豆av在线久日| 九色亚洲精品在线播放| 国产免费福利视频在线观看| 美国免费a级毛片| 夫妻性生交免费视频一级片| 亚洲国产欧美网| 成人免费观看视频高清| 啦啦啦在线免费观看视频4| 少妇被粗大的猛进出69影院| 男女边吃奶边做爰视频| 男女无遮挡免费网站观看| 在线观看免费日韩欧美大片| 麻豆乱淫一区二区| 亚洲精品久久久久久婷婷小说| 我要看黄色一级片免费的| 久久久久久久久久人人人人人人| 王馨瑶露胸无遮挡在线观看| 久久鲁丝午夜福利片| 亚洲精品久久午夜乱码| 国产伦理片在线播放av一区| 久久综合国产亚洲精品| 成人午夜精彩视频在线观看| 大片电影免费在线观看免费| 国产一区有黄有色的免费视频| 七月丁香在线播放| 成年av动漫网址| 国产一区二区三区av在线| 青春草国产在线视频| 如日韩欧美国产精品一区二区三区| 国产亚洲一区二区精品| 亚洲精品,欧美精品| 国产乱来视频区| 黄片无遮挡物在线观看| 久热久热在线精品观看| 日本午夜av视频| 免费黄网站久久成人精品| 免费播放大片免费观看视频在线观看| 亚洲国产成人一精品久久久| 日韩精品有码人妻一区| 熟妇人妻不卡中文字幕| 一区二区av电影网| 亚洲图色成人| 日日撸夜夜添|