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1.
H Li  GS Ding  CY Yue  AN Tang 《Electrophoresis》2012,33(13):2012-2018
A novel and simple method for the preparation of silica nanoparticles having surface-functionalized diamino moiety (dASNPs) was reported in our paper and characterized using scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectrometry, and thermogravimetry techniques. To test this method practically, in this contribution we describe the enhanced separation of four plant auxins - indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (dCPAA), and 2-(1-naphthyl) acetic acid (NAA) - by capillary electrochromatography using diamino moiety functionalized silica nanoparticles as pseudostationary phase (PSP) in the running buffer. The effect of pH, buffer concentration, and diamino moiety functionalized silica nanoparticles concentration on the selectivity of separation was investigated. A combination of the nanoparticles and running buffer reversed the electroosmotic direction making possible the rapid and efficient separation of the auxins from the auxins migrated in the same direction with the EOF under optimum experimental conditions. A good resolution of four auxins was obtained within 5.5 min under optimum experimental conditions. The precision (RSD, n = 5) was in the range of 0.72-0.91% and 1.89-2.23% for migration time and peak area response, respectively. The detection limits were 0.48, 0.44, 0.46, and 0.42 μM for NAA, IBA, IAA, and dCPAA, respectively. Furthermore, the method was successfully tested for the determination of IAA in the grapes.  相似文献   

2.
Li-Juan Hui  Yong Ma 《Chromatographia》2011,73(5-6):507-515
The use of silica nanoparticles with surface modification in different applications is a topic of growing interest. This article provides a systematic comparison of diamine-modified and amine-modified silica nanoparticles as pseudo-stationary phases (PSPs) in capillary electrophoresis. The different electrophoretic behavior of these PSPs was evaluated by analysis of four auxins, indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (dCPAA), and 2-(1-naphthyl)acetic acid (NAA). As expected, fast separation of the auxins was achieved, because they migrate in the same direction as the electro-osmotic flow under the optimum experimental conditions. However, the two nanoparticles had different selectivity and efficiency in the separation of the auxins, which could be related to the primary and secondary amines in their structures. The results indicated that even slight structural changes of the functional groups of the nanoparticles have pronounced effect on the interactions between the pseudo-stationary phase and the analytes.  相似文献   

3.
The low concentrations of the auxins in samples of plant tissue necessitate the use of selective and sensitive techniques for their quantification. Herein a selective and sensitive method based on dual-cloud point extraction (dCPE) and tertiary amine labeling for the quantification of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) by capillary electrophoresis-electrochemiluminescence (CE-ECL) is proposed. The procedure for dCPE included two cloud point processes with Triton X-114 as the extractant. The two auxins became hydrophobic in an acidic solution and were extracted into surfactant-rich phase after the first cloud point procedure. They were then back-extracted into the alkaline aqueous phase during the second cloud point step. The extracted auxins were reacted with 2-(2-aminoethyl)-1-methylpyrrolidine (AEMP) in acetonitrile that contained N,N′-dicyclohexylcarbodiimide and 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine to produce their AEMP-derivatives. The two auxin-AEMP-derivatives were subjected into CE and detected by Ru(bpy)32+-based ECL. The preconcentration factors for IAA and IBA with dCPE were 40.5 and 43.4, respectively. The on-capillary detection limits (S/N = 3) were 2.5 and 2.8 nM for IAA and IBA. This protocol presents a clear advantage in that it reduces the interference from the matrixes extensively and gives a high sensitivity for the detection of auxins. The proposed method was applied successfully to the detection of the two auxins in acacia tender leaves, buds, and bean sprout.  相似文献   

4.
Solid-phase microextraction followed by HPLC was used for the determination of indole-3-acetic acid (IAA), abscisic acid (ABA), indole-3-butyric acid (IBA) and 1-naphthylacetic acid (NAA) in plant samples. Parameters influencing performance, including pH, salinity, extraction time, fiber coating and temperature, were optimized. A Carbowax-coated fiber was chosen for determination due to much higher extraction efficiency compared to polyacrylate fibers. The dynamic ranges spanned over three orders of magnitude. The LOD/(LOQ) values of the target compounds in pure water were 0.149(0.497), 0.442(1.472), 0.121(0.403), 0.058(0.193) μg L−1 for IAA, ABA, IBA and NAA respectively. The method was successfully applied to the analysis of xylem fluid from Musa basjoo stem obtaining recoveries of 98.85% (IAA), 94% (IBA) and 94.30% (NAA). The method was also successfully applied to the analysis of these four target compounds in the hyperaccumulating plant, Viola baoshanensis. The results matched quite well with ones obtained by solid phase extraction followed by HPLC. The method developed was superior when applied to liquid samples because matrix effects could be eliminated.  相似文献   

5.
Zhang Y  Li Y  Hu Y  Li G  Chen Y 《Journal of chromatography. A》2010,1217(47):7337-7344
Auxin is a crucial phytohormone for precise control of growth and development of plants. Due to its low concentration in plant tissues which are rich in interfering substances, the accurate determination of auxins remains a challenge. In this paper, a new strategy for isolation and enrichment of auxins from plant tissues was obtained by the magnetic molecularly imprinted polymer (mag-MIP) beads, which were prepared by microwave heating initiated suspension polymerization using indole-3-acetic acid (IAA) as template. In order to obtain higher selective recognition cavities, an enhanced imprinting method based on binary functional monomers, 4-vinylpyridine (4-VP) and β-cyclodextrin (β-CD), was adopted for IAA imprinting. The morphological and magnetic characteristics of the mag-MIP beads were characterized by scanning electron microscopy, Fourier-transform infrared spectroscopy and vibrating sample magnetometry. A majority of resultant beads were within the size range of 80-150μm. Porous surface morphology and good magnetic property were observed. Furthermore, the mag-MIP beads fabricated with 4-VP and β-CD as binary functional monomers exhibited improved recognition ability to IAA, as compared with the mag-MIP beads prepared with the individual monomer separately. Competitive rebinding experiment results revealed that the mag-MIP beads exhibited a higher specific recognition for the template than the non-imprinted polymer (mag-NIP) beads. An extraction method by mag-MIP beads coupled with high performance liquid chromatography (HPLC) was developed for determination of IAA and indole-3-butyric acid (IBA) in plant tissues. Linear ranges for IAA and IBA were in the range of 7.00-100.0μgL(-1) and 10.0-100.0μgL(-1), and the detection limits were 3.9 and 7.4μgL(-1), respectively. The analytical performance was also estimated by seedlings or immature embryos samples from three different plant tissues, pea, rice and wheat. Recoveries were in the range of 70.1-93.5%. The results show that the present imprinting method is a promising approach for preparation of selective adsorbents for sample preparation of auxin analysis in plant tissues.  相似文献   

6.
气相色谱法直接测定植物生长素   总被引:12,自引:0,他引:12  
杜黎明  许庆琴 《色谱》2000,18(2):160-161
 建立了一种采用 5 3 0 μm大口径毛细管色谱柱、不经衍生化处理而直接测定吲哚乙酸 (IAA)、吲哚丁酸(IBA)和萘乙酸 (NAA)等植物生长素的气相色谱分析方法。以邻苯二甲酸二丁酯为内标物 ,用 FID检测 ,IAA,IBA和 NAA的相对标准偏差分别为 1 .1 4% ,0 .61 %和 0 .78%。方法简便、快速、准确、重现性好 ,可用于生长素类单组分和混合制剂的质量检测。  相似文献   

7.
A novel sample preparation method for auxin analysis in plant samples was developed by vacuum microwave-assisted extraction (VMAE) followed by molecularly imprinted clean-up procedure. The method was based on two steps. In the first one, conventional solvent extraction was replaced by VMAE for extraction of auxins from plant tissues. This step provided efficient extraction of 3-indole acetic acid (IAA) from plant with dramatically decreased extraction time, furthermore prevented auxins from degradation by creating a reduced oxygen environment under vacuum condition. In the second step, the raw extract of VMAE was further subjected to a clean-up procedure by magnetic molecularly imprinted polymer (MIP) beads. Owing to the high molecular recognition ability of the magnetic MIP beads for IAA and 3-indole-butyric acid (IBA), the two target auxins in plants can be selectively enriched and the interfering substance can be eliminated by dealing with a magnetic separation procedure. Both the VMAE and the molecularly imprinted clean-up conditions were investigated. The proposed sample preparation method was coupled with high-performance liquid chromatogram and fluorescence detection for determination of IAA and IBA in peas and rice. The detection limits obtained for IAA and IBA were 0.47 and 1.6 ng/mL and the relative standard deviation were 2.3% and 2.1%, respectively. The IAA contents in pea seeds, pea embryo, pea roots and rice seeds were determined. The recoveries were ranged from 70.0% to 85.6%. The proposed method was also applied to investigate the developmental profiles of IAA concentration in pea seeds and rice seeds during seed germination.  相似文献   

8.
Yan H  Wang F  Han D  Yang G 《The Analyst》2012,137(12):2884-2890
A highly selective molecularly imprinted solid-phase extraction (MISPE) combined with liquid chromatography-ultraviolet detection was developed for the simultaneous isolation and determination of four plant hormones including indole-3-acetic acid (IAA), indole-3-propionic acid (IPA), indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA) in banana samples. The new molecularly imprinted microspheres (MIMs) prepared by aqueous suspension polymerization using 3-hydroxy-2-naphthoic acid and 1-methylpiperazine as mimic templates performed with high selectivity and affinity for the four plant hormones, and applied as selective sorbents of solid-phase extraction could effectively eliminate the interferences of the banana matrix. Good linearity was obtained in a range of 0.04-4.00 μg g(-1) and the recoveries of the four plant hormones at three spiked levels ranged from 78.5 to 107.7% with the relative standard deviations (RSD) of less than 4.6%. The developed MISPE-HPLC protocol obviously improved the selectivity and eliminated the effect of template leakage on quantitative analysis, and could be applied for the determination of plant hormones in complicated biological samples.  相似文献   

9.
建立了同时测定水稻中吲哚-3-丁酸(IBA)、吲哚-3-乙酸(IAA)及其7种氨基酸结合物的液相色谱-串联质谱( HPLC - MS/MS)检测方法.样品在4℃下于80%甲醇中浸提12 h后,经混合阴离子交换反相固相萃取(MAX)净化,以5 mmol/L的甲酸铵溶液和甲醇为流动相,在C18柱上进行液相色谱分离,电喷雾正...  相似文献   

10.
Auxin is an important phylohormone, which regulates specific physiological responses such as division, elongation and differentiation of cells. A new method using liquid chromatography/electrospray ionization ion trap mass spectrometry (LC/ESI-ITMS) has been developed for identification and quantitation of four auxins. Under the optimum conditions, four auxins (indole-3-acetic acid, indole-3-propionic acid, indole-3-butyric acid and 1-naphthylacetic acid) were completely separated and quantitated within 7 min with a minimum detection limit of 8.0 ng mL(-1) with relative standard deviations lower than 5.0%. This method also has been applied to analysis of auxins in Chinese cabbage where, even with a complicated serious background perturbation due to the natural biological matrix, the mean recoveries ranged from 77.5% to 99.8%. Finally, we discuss the MS-relevant properties of the identified auxins in detail.  相似文献   

11.
In this work,a new sample pretreatment method prior to HPLC separations was developed for the determination of auxins in plant samples.Owing to its large surface area and high adsorption capacity, multi-walled carbon nanotube(MWCNT) was chosen as the adsorbent for the extraction of auxins from plant samples.In this study,two important auxins were selected as model analytes,namely indole-3-butyric acid(IRA) and 1-naphthylacetic acid(NAA).They could be extracted and concentrated due to theirπ-πstacking interactions with MWCNT.Then HPLC-UV was introduced to detect IBA and NAA after sample pretreatment.Factors that may affect the enrichment efficiency were investigated and optimized.Comparative studies showed that MWCNT was superior to C18 for the extraction of the two analytes.Validation experiments showed that the optimized method had good linearity(0.9998 and 0.9960),high recovery(81.4%-85.4%),and low detection limits(0.0030 mg/L and 0.0012 mg/L).The results indicated that the novel method had advantages of convenience,good sensitivity,high efficiency, and it was feasible for the determination of auxins in plant samples.  相似文献   

12.
A new, simple and rapid capillary electrophoresis (CE) method, using hexadimethrine bromide (HDB) as electroosmotic flow (EOF) modifier, was developed for the identification and quantitative determination of four plant hormones, including gibberellin A3 (GA3), indole-3-acetic acid (IAA), alpha-naphthaleneacetic acid (NAA) and 4-chlorophenoxyacetic acid (4-CA). The optimum separation was achieved with 20 mM borate buffer at pH 10.00 containing 0.005% (w/v) of HDB. The applied voltage was -25 kV and the capillary temperature was kept constant at 25 degrees C. Salicylic acid was used as internal standard for quantification. The calibration dependencies exhibited good linearity within the ratios of the concentrations of standard samples and internal standard and the ratios of the peak areas of samples and internal standard. The correlation coefficients were from 0.9952 to 0.9997. The relative standard deviations of migration times and peak areas were < 1.93 and 6.84%, respectively. The effects of buffer pH, the concentration of HDB and the voltage on the resolution were studied systematically. By this method, the contents of plant hormone in biofertilizer were successfully determined within 7 min, with satisfactory repeatability and recovery.  相似文献   

13.
The separation and determination of proteins in food is an important aspect in food industry. Inspired by the self‐polymerization of dopamine under alkaline conditions and the natural adhesive properties of polydopamine, in this paper, a simple and economical method was developed for the preparation of polydopamine‐coated open tubular column, in which ammonium persulfate was used as the source of oxygen to induce and facilitate the polymerization of dopamine to form polydopamine. In comparison with a naked fused‐silica capillary, the direction and magnitude of the electro‐osmotic flow of the as‐prepared polydopamine‐coated open tubular column could be manipulated by varying the pH values of background solutions due to the existence of amine and phenolic hydroxyl groups on polydopamine coating. The surface morphology of the polydopamine‐coated open tubular column was studied by scanning electron microscopy, and the thickness of polydopamine coating was 106 nm. The performance of the polydopamine‐coated open tubular column was validated by analysis of proteins. The relative standard deviations of migration times of proteins representing run‐to‐run, day‐to‐day, and column‐to‐column were less than 3.5%. In addition, the feasibility of the polydopamine‐coated open tubular column for real samples was verified by the separation of proteins in chicken egg white and pure milk.  相似文献   

14.
建立了同时检测水稻中6种内源性植物激素脱落酸( Abscisic acid,ABA)、吲哚-3-乙酸( Indole-3-acetic acid, IAA)、水杨酸( Salicylic acid,SA)、茉莉酸( Jasmonic acid,JA)、吲哚-3-丙酸( Indole-3-propionic acid, IPA)和吲哚-3-丁酸( Indole-3-butyric acid,IBA)的全自动在线固相萃取-液相色谱-串联质谱方法。植物样品经过甲醇提取,采用C18固相萃取柱富集净化,流动相将待测物洗脱至C18分析色谱柱进行分离,最终使用串联四极杆质谱进行检测。方法的线性范围为8~320μg/L,相关系数为R2≥0.99;方法的检出限(S/N=3)范围为0.1~0.8μg/kg;实际样品中方法回收率范围为71.2%~126%,RSD<13%。应用本方法快速、准确地检测了水稻幼穗中多种内源性植物激素的含量,并与目前植物学领域内常用的检测方法进行了比较。同时,本方法对水稻受伤叶片的内源植物激素含量变化进行了定量分析,其含量随受伤时间的变化趋势与其生物背景的实验结果相吻合。  相似文献   

15.
张淼  王雨晨  MUHAMMADAtif  陈丽娟  王延梅 《色谱》2020,38(9):1085-1094
制备了一种对溶菌酶具有可控吸附性能的混合刷涂层毛细管,用于毛细管电泳在线富集溶菌酶以提高其检测灵敏度。首先,分别通过阳离子开环聚合和可逆加成-断裂链转移(RAFT)聚合合成聚(2-甲基-2-噁唑啉)(PMOXA)和聚丙烯酸(PAA),然后将甲基丙烯酸缩水甘油酯(GMA)分别与PMOXA和PAA通过自由基共聚和RAFT聚合合成出聚(2-甲基-2-噁唑啉)-r-甲基丙烯酸缩水甘油酯(PMOXA-r-GMA)和聚丙烯酸-b-聚甲基丙烯酸缩水甘油酯(PAA-b-PGMA)。将PMOXA-r-GMA和PAA-b-PGMA的混合溶液以一定比例加入到毛细管内,通过加热即可制备出基于PMOXA和PAA的混合刷涂层毛细管。X射线光电子能谱(XPS)对毛细管原材料的表面组成研究结果表明,当混合溶液质量浓度为20 g/L、PMOXA-r-GMA和PAA-b-PGMA质量比为1:1时,所得涂层中羧基的含量随着PAA链长的增加而增加;异硫氰酸荧光素标记溶菌酶(FITC-溶菌酶)吸附实验结果显示,通过改变环境的pH和离子强度(I)可以调控涂层毛细管对溶菌酶的吸附和释放,在pH 7(I=10-5mol/L)条件下,毛细管可以吸附大量的溶菌酶,当条件变为pH 3(I=10-1mol/L)时,吸附的溶菌酶可以被释放出来。将这种具有溶菌酶可控吸附性能的涂层毛细管用于毛细管电泳在线富集溶菌酶,当PAA链长是PMOXA链长的2.2倍时,溶菌酶的灵敏度增强因子为17.69,检出限为8.7×10-5g/L;同一天内对溶菌酶连续测定5次以及连续测定5天,峰面积的日内、日间相对标准偏差(RSD)分别为2.9%和4.1%,迁移时间的日内、日间RSD分别为0.9%和2.1%。涂层的制备只需一步,简单易行,而且涂层具有很好的稳定性。本研究为毛细管电泳分析痕量蛋白质提供了一种简单有效的方法。  相似文献   

16.
Based on the self-polymerization and strong adhesion characteristics of dopamine in aqueous solution, a novel and convenient approach was developed to immobilize protein onto porous polyethylene (PE) membranes. A thin polydopamine (pDA) layer was formed and tightly coated onto PE membrane by dipping simply the membrane into dopamine aqueous solution for a period of time. Subsequently, bovine serum albumin (BSA) was bound onto the obtained PE/pDA composite membranes via the coupling between BSA and the reactive polydopamine layer. The firm immobilization of polydopamine layer and BSA was verified by attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). The results of water contact angle measurement showed that the hydrophilicity of PE membrane was significantly improved after coating polydopamine and binding BSA. The experiments of blood platelet adhesion indicated that BSA-immobilized PE membrane had better blood compatibility than the unmodified PE and the PE/pDA composite membranes. The investigations on hepatocyte cultures and cell viability revealed that the polydopamine coating endowed PE membrane with significantly improved cell compatibility. Compared to BSA surface, polydopamine surface is more favorable for cell adhesion, growth, and proliferation.  相似文献   

17.
In recent years, nanoparticles have gained more attention when used in separation science. In this study, chitosan‐modified silica nanoparticles were successfully synthesized and characterized by transmission electron microscopy, elemental analysis and zeta potential measurements, etc. When added into the running buffer solution as pseudo‐stationary phase in capillary electrophoresis, the separation of four representative auxins, i.e., indole‐3‐acetic acid, indole butyric acid, 2,4‐dichlorophenoxyacetic acid, 1‐naphthaleneacetic acid, was carried out. Some important factors, such as the nanoparticles concentration, the pH and concentration of the running buffer solution, were also investigated on the separation. Under optimized experimental conditions, all the auxins investigated can be baseline separated within 5 min with higher column performance. The method established can also be used for quantitative analysis. The relative standard deviations obtained for indole‐3‐acetic acid, indole butyric acid, 2,4‐dichlorophenoxyacetic acid, 1‐naphthaleneacetic acid were in the range of 1.6–5.7% for peak area and 0.53–1.60% for migration time. The calibration curves obtained from the peaks areas for auxins were linear in the range of 0.1–80 mg/L with the correlation coefficients of 0.994–0.999. The limit of detection (S/N = 3) was 11–75 μg/L. The developed method was also successfully used for the determination of auxins in fruits and vegetables samples with good recoveries.  相似文献   

18.
A rapid, simple, and efficient method for the fast determination of multiple phytohormones was developed in this work, based on single-drop liquid-liquid-liquid microextraction (SD-LLLME) combined with direct analysis in real-time mass spectrometry (DART-MS). Six phytohormones--indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), jasmonic acid (JA), salicylic acid (SA), abscisic acid (ABA), and gibberellin A(3) (GA(3))--were analyzed simultaneously using this method, and the conditions employed for DART-MS and SD-LLLME were optimized systematically. Satisfactory results were obtained in terms of linearity (R (2) values for all phytohormones were 0.991-0.996), sensitivity (limits of detection were 0.65-72 ng/mL), and repeatability (RSD values were 6.9-14%). In addition, the proposed method was applied to determine the endogenous phytohormones in three kinds of fruit juice. Different concentrations of phytohormones were detected with satisfactory recoveries, and the whole analytical procedure took no more than 30 min. Therefore, this combination of SD-LLLME and DART-MS was shown to be a suitable and effective approach for the fast analysis of targets present at trace level concentrations in complex matrices.  相似文献   

19.
In this work, a binary-mixed-brushes-coated (BBC) capillary with switchable protein adsorption/desorption properties was developed and applied for on-line preconcentration of proteins. Firstly, amine-terminated poly(2-methyl-2-oxazoline) (PMOXA-NH2) and thiolterminated poly(acrylic acid) (PAA-SH) were synthesized by using cationic ring-opening polymerization (CROP) and reversible addition fragmentation chain transfer (RAFT) polymerization, respectively. Then, the BBC capillary based on poly(2-methyl-2-oxazoline) (PMOXA) and poly(acrylic acid) (PAA) was prepared by sequentially grafting of PMOXA-NH2 and PAA-SH onto fused-silica capillary inner surface through poly(dopamine) (PDA) as an anchor. The obtained PMOXA/PAA coating formed on the capillary or capillary's raw material was characterized in terms of the thickness, surface chemical composition by using scanning electron microscope (SEM) and X-ray photoelectron spectrum (XPS). The switchable protein adsorption/desorption performance of the BBC capillary was investigated by using fluorescence microscope under di erent solutions with certain pH and ionic strength(I). The results showed that bovine serum albumin (BSA) could be adsorbed on BBC capillary at pH=5.0 (I=10-5 mol/L), and then the adsorbed BSA could be released at pH=9.0 (I=0.1 mol/L). This switchable protein adsorption/desorption property of coated capillary was then used to preconcentrate proteins on-line for increasing the detection sensitivity of BSA in capillary electrophoresis (CE). With this method, a sensitivity enhancement factor (SEF) more than 5000 for BSA detection was obtained.  相似文献   

20.
In this work, we developed a capillary column modified with zeolitic imidazolate framework‐8 as a novel stationary phase for open‐tubular capillary electrochromatography. To immobilize zeolitic imidazolate framework‐8 onto the inner surface of silica capillary, a bio‐inspired polydopamine functionalization was used to functionalize the capillary surface with polydopamine. First, a polydopamine layer was assembled inside the capillary. Second, due to noncovalent adsorption and covalent reaction ability, polydopamine could attract and anchor zeolitic imidazolate framework‐8 onto the inner surface of capillary. It has been demonstrated that zeolitic imidazolate framework‐8 was successfully grafted on the inner wall of the capillary by scanning electron microscopy, and Fourier transform infrared spectroscopy. The electro‐osmotic flow characteristics of capillaries were also investigated by varying the pH value and acetonitrile content of mobile phase. The zeolitic imidazolate framework‐8 coating not only increased the phase ratio of open‐tubular column, but also improved the interactions between tested analytes and the stationary phase. Three groups of isomers including acidic, basic, and neutral compounds were well separated on the zeolitic imidazolate framework‐8 bonded column, with theoretic plate numbers up to 1.9 × 105 N for catechol. The repeatability of the prepared columns was also studied, and the relative standard deviations for intra‐ and interday runs were less than 5%.  相似文献   

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