#dionex ion chromatography

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Global High Performance Liquid Chromatography (HPLC) Market 2019 - Partition Chromatography

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Structure and functional dynamics of the mitochondrial Fe/S cluster synthesis complex

Protein expression and purification for crystallography

The plasmid pZM2 harboring human NFS1 encoding residues 56–457 (∆1-55) with a N-terminal His6-tag and plasmid pZM4 containing full-length ISD11 (no tag) were gifts from Silke Leimkühler (Molekulare Enzymologie, Universität Potsdam, Germany; Supplementary Table 3). The NFS1 and ISD11 genes were co-expressed in One Shot BL21 Star (DE3) cells (ThermoFisher Scientific) by selection on solid LB-Agar media containing ampicillin (100 µg ml−1 and chloramphenicol (30 µg ml−1). An overnight culture (6 ml, 37 °C) was used to inoculate 2 L of Terrific Broth (TB) medium (Fisher) containing the two selection antibiotics and supplemented with 10 µM PLP (Sigma-Aldrich) and 10 µM FeSO4. After growth at 37 °C until OD600 0.6–0.8, the temperature was lowered to 22 °C. Protein expression was induced by 1 mM IPTG and cultured overnight. For expression of ISCU, the plasmid p24ac (gift from Dr. Kuanyu Li, Nanjing University, China; Supplementary Table 3) was used encoding ISCU1 residues 2–142 with a C-terminal His6-tag50. The protein sequence was altered in position 107 to yield ISCU1-M107I protein (termed ISCU; plasmid p24ISCU_MI) that was described before as FXN-independent32, in the expectation that frataxin would be dispensable as part of the core ISC complex. Cells were harvested by centrifugation for 20 min at 9000×g. The cell pellet was resuspended in purification buffer (10 mM BIS-TRIS pH 5.5, 200 mM NaCl, 20 mM KCl, 2 mM NaH2PO4, 2 mM Na2HPO4, 5% glycerol) and flash frozen in liquid nitrogen for storage at −80 °C.

The cell pellet containing co-expressed NFS1 and ISD11 was thawed at 42 °C and supplemented with 0.1 mM phenylmethylsulfonyl fluoride and 10 µM PLP. Resuspended cells were disrupted using the Constant Cell Disruption System (Constant System LTD., UK) under 35 MPa pressure. Cell lysate was spun down for 30 min at 20,000×g at 4 °C in Beckman Avanti J-26XP centrifuge. The supernatant was loaded on a Ni-NTA column (GE Healthcare) using Bio-Rad NGC FPLC system with an integrated sample pump. The column was washed with 20 column volumes of purification buffer supplemented with imidazole in a 0–75 mM gradient. The proteins were eluted with 2 column volumes of 125 mM imidazole in purification buffer containing 1 mM 1,4-dithiothreitol (DTT). Soluble NFS1-ISD11 complex was eluted as a single peak as followed by absorbance at 280 nm (aromatic amino acid residues) and 420 nm (PLP). Mass spectrometry of the purified protein fraction indicated, in addition to NFS1 and ISD11, the unexpected presence of E. coli ACP. The human NFS1-ISD11-ACP complex (termed (NIA)2) was concentrated to 12–15 mg ml−1, vitrified in liquid nitrogen, and stored in −80 °C.

To purify the human NFS1-ISD11-ACP-ISCU complex (termed (NIAU)2), plasmid pZM2, encoding NFS1 with N-terminal His6-tag, was modified to yield a NFS1 construct (56–457) without tag (Supplementary Tables 3 and 4). Proteins NFS1-ISD11 (no tag) and ISCU-His6 were expressed in separate cells, and pellets were combined prior to purification, which followed the standard protocol for isolation of the (NIA)2 complex. The (NIAU)2 complex was eluted from the Ni-NTA column with 75 mM imidazole in purification buffer, and was immediately supplemented with 1 mM DTT and with or without 1 mM EDTA. Proteins were concentrated to 17–20 mg ml−1 and vitrified in liquid nitrogen.

Dynamic Light Scattering and Multi Angle Light Scattering

Prior to crystallization experiments the polydispersity of the human (NIA)2 complex in solution was measured using Wyatt DLS Plate Reader II at 826.7 nm at a range of temperatures. The sample and buffer were pre-filtered using 0.22 μm syringe filter and 50 μl of solution was injected into a measurement well in 384-well plate from Greiner Bio-One (Monroe, NC, USA). The average polydispersity (%Pd) of the purified (NIA)2 complex was calculated using Dynamics V7 software. The best results were obtained at temperatures around 12 °C and the estimated polydispersity was in the 15–20% range. The average molecular masses of the complexes were determined using a multi-angle light scattering using size exclusion chromatography column (Wyatt SEC WTC 030S5) connected to GE Healthcare FPLC ÄKTA system in tandem with miniDAWN TREOS and Wyatt refractometer Optilab tREX. Fractions were collected and protein purity was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Molecular mass of the human (NIA)2 complex estimated as 125–130 kDa using Wyatt ASTRA 6 software, which suggested a dimeric state (calculated MMth = 146.7 kDa). The (NIAU)2 complex was subjected to the crystallization screening directly after size exclusion chromatography, without evaluating polydispersity.

Circular Dichroism

The secondary structure and folding of the (NIA)2 complex was investigated by CD using Chirascan Plus CD Spectrometer (Applied Photophysics, UK). Purified complex was diluted into buffer (10 mM BIS-TRIS pH 6, 50 mM NaCl), and the CD spectrum was measured between 280–195 nm at 20 °C. The spectrum was analyzed using Chirascan software to determine the secondary structure content. CD was also used to measure thermal stability of the complex. Spectra were measured at temperatures from 5–90 °C and analyzed with the Global3 software yielding a melting temperature of the complex between 25–30 °C.

Mass spectrometry of gel fragments containing proteins

Mass spectrometry analysis of the proteins separated on 15% SDS-PAGE gel was performed as follows. Gel fragments containing proteins were resuspended in 20 µL trypsin buffer (trypsin in 1 mM HCl and 200 mM NH4HCO3) followed by addition of 30 µL of 200 mM NH4HCO3 to each sample. Samples were incubated at 30 °C with shaking at 300 rpm. Trypsin digestion reaction was quenched by adding 1% trifluoroacetic acid (TFA) and tryptic peptides were extracted from gel samples using 100 µL of 0.1% TFA in 60% acetonitrile (ACN) and dried in the speed vac. Peptides were reconstituted in 10 µL of MS grade water:ACN:formic acid (97:3:0.1 v v−1) and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Spectral results were collected over a mass range of 250–1700 (mass*charge−1; m z−1) at a scan rate of 8 spectra per s. MS/MS data were collected over a range of 50–1700 m z−1 and a set isolation width of 1.3 atomic mass units. Spectral data were converted to a mass*charge−1 data format using Agilent MassHunter Qualitative Analysis Software (Agilent Technologies Canada Ltd., Mississauga, ON, CA) and were processed against the UniProt Escherichia coli database, using SpectrumMill (Agilent Technologies Canada Ltd., Mississauga, ON, CA) as the database search engine. Search parameters included a fragment mass error of 50 ppm, a parent mass error of 20 ppm, trypsin cleavage specificity, and carbamidomethyl as a fixed modification of cysteine and oxidized methionine as a variable modification.

Mass spectrometry of proteins in solution

Protein solutions containing purified (NIA)2 or (NI)2 were digested by the addition of Sequencing Grade Modified Trypsin (Promega) and incubated at 37 °C overnight. The mass spectrometric analysis of the samples was performed using an Orbitrap Velos Pro mass spectrometer (ThermoScientific). An Ultimate nanoRSLC-HPLC system (Dionex), equipped with a custom 20 cm × 75 µm C18 RP column filled with 1.7 µm beads was connected online to the mass spectrometer through a Proxeon nanospray source. 1–15 µL of the tryptic digest (depending on sample concentration) were injected onto a C18 pre-concentration column. Automated trapping and desalting of the sample was performed at a flow rate of 6 µL min−1 using water/0.05% formic acid as solvent. Separation of the tryptic peptides was achieved with the following gradient of water/0.05% formic acid (solvent A) and 80% ACN/0.045% formic acid (solvent B) at a flow rate of 300 nL min−1: holding 4% B for 5 min, followed by a linear gradient to 45%B within 30 min and linear increase to 95% solvent B in additional 5 min. The column was connected to a stainless steel nanoemitter (Proxeon, Denmark)) and the eluent was sprayed directly towards the heated capillary of the mass spectrometer using a potential of 2300 V. A survey scan with a resolution of 60,000 within the Orbitrap mass analyzer was combined with at least three data-dependent MS/MS scans with dynamic exclusion for 30 s either using CID with the linear ion-trap or using HCD combined with orbitrap detection at a resolution of 7500. Data analysis was performed using Proteome Discoverer (ThermoScientific) with SEQUEST and MASCOT (version 2.2; Matrix science) search engines using either SwissProt or NCBI databases. For quantitation of the protein abundance the “integrated peak area” of the spectra was used.

Protein crystallization

The crystallization screens for the (NIA)2 complex were dispensed into 96-well sitting drop plate using Gryphon crystallization robot (ArtRobbins Instruments, Sunnyvale, CA). Each drop contained 300 nL of the complex concentrated to 12 mg ml−1 and 300 nL of the mother liquor. The initial crystals appeared at 12 °C in a well containing the Protein Complex suite (Qiagen, Toronto) solution 100 mM HEPES pH 7.5, 0.2 M ammonium acetate and 25% isopropanol. Optimized crystals were obtained from the wells containing 100 mM 2-(N-morpholino)ethanesulfonic acid (MES) pH 6.5, 0.3 M ammonium acetate, 20 mM calcium acetate, 20 mM CaCl2, and 19% isopropanol. The crystallization plate was transferred to 4 °C, and the crystals were harvested immediately and stored in liquid nitrogen using 17% MPD as a cryo-protectant. The best crystals diffracted to 2.75 Å resolution, belonged to the trigonal system, space group R32, with cell dimensions a = 140.8 Å, b = 140.8 Å, c = 203.3 Å, α = β = 90˚, γ = 120˚, and contained one molecule of each protein in the asymmetric unit.

The (NIAU)2 complex solution was concentrated to 17–20 mg ml−1 and dispensed into 96-well sitting drop plate as described above. The initial crystals appeared at 20 °C in wells containing 0.1 M MES pH 6.5, 10% PEG-20,000 or 0.1 M HEPES pH 7.5, 20% PEG-8000 in Compas Suite (Qiagen, Toronto). The best diffracting crystals were grown in the optimized condition of 0.1 M MES pH 7.0, 15% PEG-3350 at 15 °C to initialize nucleation. After 3 h plates were transferred to 12 °C, and incubated until crystals were fully-grown. Crystals were harvested at 4 °C and stored in liquid nitrogen using 20% glycerol as a cryo-protectant. Crystals with Zn ((NIAU-Zn)2 complex) diffracted to 3.3 Å, space group P212121 with a = 97.5 Å, b = 121.2 Å, c = 151.5 Å, and without Zn ((NIAU)2 complex) to 3.15 Å, space group P212121 with cell dimensions a = 98.4 Å, b = 123.3 Å, c = 151.7 Å.

Data collection and structure refinement

Diffraction data were collected using 08ID beamline at the Canadian Light Source, CMCF Sector, Saskatoon, SK, using a Rayonix MX300 CCD detector. Data were integrated and scaled using XDS package51. Initial phases were derived by molecular replacement method with Phaser MR52 using E. coli IscS cysteine desulfurase (PDB code 3LVM)16 as a model. The ISD11 model was built into density using ARP/wARP53. Additional electron density was present in the map calculated at this stage. Based on the mass spectrometric identification of ACP as part of the complex the MolRep program54 was used to place the E. coli ACP model (PDB code 2FAE,31 and it fitted nicely into the unaccounted density. The structure was refined using the PHENIX package55, and manual rebuilding was carried with COOT56. A strong density in the 2mFo-DFc and difference maps extended from the residue Ser36 of ACP in all structures and was modeled as a lipid molecule with a chain containing 12 carbon atoms. Similarly, the PLP molecule was clearly visible in all three structures and was included in the model.

The final model of the (NIA)2 complex contains residues 65–84, 97–274, 295–359, 404–431 of NFS1, 3–77 of ISD11, and 3–73 of ACP. Three segments of NFS1 are disordered in the structure, namely 85–96, 275–294, and 360–403, the latter containing the active-site cysteine (Cys loop) (Supplementary Fig. 1d). The final Rwork and Rfree are 0.213 and 0.255. The model of (NIAU-Zn)2 contains NFS1 residues 54–452 (chains A), 54–454 (chain E), ISD11 5–85 (chain B) and 3–85 (chain F), ACP 4–74 (chain C), and 3–72 (chain G), ISCU 6–133 (chain D), and 10–133 (chain H). The NFS1 chain E has a partially disordered Cys loop for residues Cys381-Ser383. The final Rwork and Rfree are 0.196 and 0.255. The (NIAU)2 complex showed more distortion of the NFS1 chain A (C terminus). Both NFS1 chains lack residues of the flexible Cys loop (Ala381-Leu385) due to the lack of stabilization by Zn ions bound to ISCU. The ISCU chain D is less well ordered and while the main chain trace is sufficiently well defined, many side chains are only partially ordered and could not be reliably modeled. This model contains NFS1 residues 55–380 and 386–455 (chains A), 54–379 and 385–456 (chain E), ISD11 5–85 (chain B) and 4–85 (chain F), ACP 4–75 (chain C) and 3–73 (chain G), ISCU 15–132 (chain D) and 10–135 (chain H). The final Rwork and Rfree are 0.188 and 0.242. The model geometries were verified by MolProbity57. Data collection and refinement statistics are provided in Table 1.

Table 1: X-Ray diffraction data and refinement statistics

Protein expression and purification for biochemistry or SAXS

CtNfs1-CtIsd11(-His10)-ACP, CtIsu1-His6, or His6-CtYfh1 from C. thermophilum, and the NFS1-ISD11(-His10) complex from H. sapiens were expressed in E. coli and purified by Ni-Sepharose affinity chromatography and gel filtration as described previously28. Variations in liquid media led to different amounts of bound ACP. Human or C. thermophilum NFS1-ISD11(-His10) complexes purified from cells grown in LB medium did not bind any detectable amounts of ACP, while purification from cells grown in TB medium yielded substantial amounts of bound ACP. This is likely due to different lipid chain lengths. Recombinant human FDX227 and ISCU2 (Supplementary Table 3) was purified by anion and cation exchange chromatography, respectively, and subsequent gel filtration as described previously.

Reconstitution of de novo Fe/S cluster synthesis on ISCU

In vitro reconstitution assays6, 28 were prepared in a Coy anaerobic chamber using freshly prepared stock solutions. Protein solutions were stored under anaerobic conditions for at least 6 h prior to experiments. The 300 µL standard reaction contained 2.5 µM C. thermophilum or human NFS1-ISD11±ACP, 75 µM ISCU2, 3 µM FXN, 3 µM FDX2, 0.3 µM FdxR in reconstitution buffer (35 mM Tris pH 8.0, 150 mM NaCl, 200 µM MgCl2, 300 µM FeCl2, 800 µM Na-ascorbate, and 500 µM NADPH). The reaction was transferred to a CD cuvette, sealed tightly and incubated at 30 °C in a CD spectrometer (Jasco, J-815). The CD-signal at 431 nm was recorded and after 2 min the reconstitution reaction was initiated by the addition of 500 µM cysteine. After 20 min a full spectrum was recorded from 300 nm to 650 nm.

Protein interactions by Microscale Thermophoresis

MST58 was performed on a Monolith NT.115 (Nano Temper Technologies GmbH, Munich, Germany) at 21 °C (red LED power was set to 50% and infrared laser power to 75%). 20 µM human (NIA)2 complex was labeled with the dye NT 647 supplied by Nano Temper Technologies. Labeled (NIA)2 complex was titrated as indicated with ISCU2 in buffer T (35 mM KPi pH 7.4 and 150 mM NaCl). Prior to MST measurements ISCU2 was treated with 5 mM DTT, 1 mM KCN, and 10 mM EDTA and subsequently gel filtered using a HiLoad 16/60 Superdex 75 PG (GE Healthcare) either aerobically or anaerobically in a Coy anaerobic chamber. At least nine independent MST experiments were performed at 680 nm and processed by Nano Temper Analysis package 1.2.009 and Origin8 (OriginLab, Northampton, MA).

Dissociation of the NFS1-ISD11-ACP complex

25 µM CtNfs1-CtIsd11-His10-ACP in buffer D (35 mM Tris-HCl pH 7.4, 150 mM NaCl and 5% (v v−1) glycerol) was anaerobically incubated for 4 h at 15 °C with either 15 mM cysteine or 15 mM serine or 3 mM DTT. Samples were bound to Ni-NTA agarose (IBA Life Science) and incubated aerobically at 4 °C for 1 h. Samples were centrifuged at 1500×g at 4 °C for 5 min, and the supernatants were analyzed by SDS-PAGE and Coomassie staining. The remaining Ni-NTA beads were incubated for 10 min with buffer D containing 250 mM imidazole, and again centrifuged as described above. The supernatants were analyzed as above.

SAXS methodology and modeling of components in complexes

All complexes and monomeric proteins were prepared or mixed in buffer S1 (35 mM Tris-HCl pH 7.4, 150 mM NaCl with or without 5 mM Cys) in a Coy anaerobic chamber. Protein solutions were subsequently subjected to size exclusion chromatography. Prior to final measurements monomeric ISC proteins and the various ISC complexes were desalted twice by passing them through a PD10 column (GE Healthcare) in buffer S1. Samples were subsequently cleared by ultracentrifugation (20,000×g, 20 min) and placed in the sample holder. SAXS data were collected at minimum three protein concentrations ranging from 1 to 10 mg ml−1. The SAXS experiments were carried out on the BM29 BioSAXS beamline at ESRF (European Synchrotron Radiation Facility) equipped with a Pilatus 1 M detector at standard set-up. Ten frames were recorded for each concentration and sample.

SAXS data evaluation was carried out using the ATSAS software package (ATSAS 2.6.0)59. GNOM60 was used to obtain the Pair distribution function (P r ) and D max values. The radius of gyration (R g ) was determined using the Guinier approximation. For calculation of 20 ab initio models the output of GNOM was used together with DAMMIF61 and the respective 3D structures were subsequently fitted to the average shape using UCSF Chimera 1.1162. The location and orientation of CtFdx2 was adjusted to the known interaction surface within the ferredoxin-Isu1 heterodimer28. The location and orientation of CtYfh1 relative to CtNfs1 was based on NMR and SAXS solution structures of the bacterial counterparts (IscS-CyaY36) and the location of bacterial CyaY on IscS16. For final rigid body modeling of all proteins/complexes, SASREF was used63. Therefore, scattering amplitudes for human ISC complexes were calculated based on our crystal structures using CRYSOL64 and compared to experimental data. For the C. thermophilum ISC proteins and complexes, we used homology models based on E. coli or yeast structures. All samples yielded good or excellent χ2 values.

Analysis of ISD11 mutations in yeast and enzyme activities

Wild-type and mutant versions of S. cerevisiae ISD11 (nucleotides 1–375, synthesized by GenScript, Piscataway, USA) with a C-terminal FLAG-tag were inserted into vector p416MET25 (Supplementary Table 3). S. cerevisiae GalL-ISD11 cells (W303-1A; pISD11::pGALL-natNT2; this work) harboring the different versions of p416-ISD11-FLAG and pRS414-ACP1-HA25 (Supplementary Table 3) were cultivated in SD minimal medium for 40 h and mitochondria were isolated65. Mitochondria were lysed at a protein concentration of 1.5 µg µl−1 for 5 min at 4 °C in buffer E (50 mM Tris-HCl (pH 8), 50 mM NaCl, 0.2% (v v−1) Triton-X100). Debris was removed by centrifugation (20,000 xg, 20 min at 4 °C). Enzyme activities were measured according to Pierik et al.66. Desulfurase activity determination of NFS1 was carried out by detecting sulfide production with methylene blue67 using 20 µg of C. thermophilum or human purified NFS1-ISD11 with or without bound ACP. For determination of desulfurase activity in isolated mitochondria, 200 µg of mitochondrial lysates (see above) were used.

Co-immunoprecipitation of mitochondrial proteins

Mitochondria (250 µg of protein) were lysed for 5 min on ice in 250 µl buffer (10 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.1 mM DTT, 10 % v v−1 glycerol, 0.2% v v−1 Triton-X100) supplemented with 2 mM phenylmethylsulphonylfluoride. Membrane debris was removed by centrifugation (12000×g, 15 min, 4 °C) and the mitochondrial lysate was incubated with 30 µl FLAG-M2 Sepharose beads (Sigma Aldrich) for 1 h on a rotary shaker at 4 °C. Beads were pelleted by centrifugation (1000×g, 1 min, 4 °C) and washed 3 times with 500 µL buffer. Bound proteins were subjected to SDS polyacrylamide gel electrophoresis and identified by immunostaining using specific antibodies (α-Nfs1, α-Isu1, and α-porin: self-raised (1:5000; rabbit), α-HA: Santa Cruz (sc-7392) (1:15,000; mouse); α-FLAG: SIGMA (011M4789) (1:20,000; rabbit); secondary α-mouse: Bio-Rad (L1706516) (1:1000); secondary α-rabbit: Sigma-Aldrich (A1949) (1:1000)).

Miscellaneous methods

The following published methods were used: yeast cell technologies68; manipulation of DNA and PCR69; and immunological techniques70. Plasmids used in this study are listed in Supplementary Table 3. Primers used in this study are listed in Supplementary Table 4.

Data availability

Atomic coordinates and structure factors for ((NIA)2)2, ((NIAU)2)2, and (NIAU-Zn)2 complexes were deposited at the Protein Data Bank (http://www.rcsb.org/pdb/home/home.do) with the codes 5WGB, 5WKP, and 5WLW, respectively. The data that support the findings of this study are available from the corresponding authors upon reasonable request.

— Nature Communications

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Dionex CD25 Conductivity Detector

Conductivity Detector CD25 Dionex

Dionex Conductivity Detector CD25. Detects and quantifies ionic analytes in liquid and  ion chromatography. Range;0.01µS to    3000 uS, full scale.

[kimberlabtron]

Ion Chromatograph LPIC-A10 is a light weight, portable design model with powerful data processing system. The quick ion chromatograph columns are used for rapid 5 min detection. Adopted with built –in multi-channel solvent selection valve helps in gradient elusion, it shortens peak time, and improves efficiency of the chromatograph conducted. Upgraded with dual detection system (dual column for various applications), supports the needs of different industries.

[abhigunjal]

High Performance Liquid Chromatography

Wiseguyreports.Com Adds “High Performance Liquid Chromatography Market -Market Demand, Growth, Opportunities, Manufacturers, Analysis of Top Key Players and Forecast to 2022” To Its Research Database.

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 Covered In This Report:

In this report, the global High Performance Liquid Chromatography (HPLC) market is valued at USD XX million in 2016 and is expected to reach USD XX million by the end of 2022, growing at a CAGR of XX% between 2016 and 2022.

Geographically, this report is segmented into several key Regions, with production, consumption, revenue (million USD), market share and growth rate of High Performance Liquid Chromatography (HPLC) in these regions, from 2012 to 2022 (forecast), covering North America Europe China Japan Southeast Asia India Global High Performance Liquid Chromatography (HPLC) market competition by top manufacturers, with production, price, revenue (value) and market share for each manufacturer; the top players including Thermo Fisher Scientific Waters Shimadzu Agilent Technologies Dionex PerkinElmer Zeiss GE Healthcare Linde-gas (HiQ) Sharp Air Products Gilson Buck Scientific Sigma-Aldrich Bio-Rad Sunny Optical Technology Jasco Phenomenex On the basis of product, this report displays the production, revenue, price, market share and growth rate of each type, primarily split into Partition Chromatography Normal-phase Chromatography Displacement Chromatography Reversed-phase Chromatography (RPC) Size-exclusion Chromatography Ion-exchange Chromatography Bioaffinity Chromatography On the basis on the end users/applications, this report focuses on the status and outlook for major applications/end users, consumption (sales), market share and growth rate of High Performance Liquid Chromatography (HPLC) for each application, including Manufacturing Legal Research Medical

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Table of Contents

Global High Performance Liquid Chromatography (HPLC) Market Research Report 2017 1 High Performance Liquid Chromatography (HPLC) Market Overview 1.1 Product Overview and Scope of High Performance Liquid Chromatography (HPLC) 1.2 High Performance Liquid Chromatography (HPLC) Segment by Type (Product Category) 1.2.1 Global High Performance Liquid Chromatography (HPLC) Production and CAGR (%) Comparison by Type (Product Category) (2012-2022) 1.2.2 Global High Performance Liquid Chromatography (HPLC) Production Market Share by Type (Product Category) in 2016 1.2.3 Partition Chromatography 1.2.4 Normal-phase Chromatography 1.2.5 Displacement Chromatography 1.2.6 Reversed-phase Chromatography (RPC) 1.2.7 Size-exclusion Chromatography 1.2.8 Ion-exchange Chromatography 1.2.9 Bioaffinity Chromatography 1.3 Global High Performance Liquid Chromatography (HPLC) Segment by Application 1.3.1 High Performance Liquid Chromatography (HPLC) Consumption (Sales) Comparison by Application (2012-2022) 1.3.2 Manufacturing 1.3.3 Legal 1.3.4 Research 1.3.5 Medical 1.4 Global High Performance Liquid Chromatography (HPLC) Market by Region (2012-2022) 1.4.1 Global High Performance Liquid Chromatography (HPLC) Market Size (Value) and CAGR (%) Comparison by Region (2012-2022) 1.4.2 North America Status and Prospect (2012-2022) 1.4.3 Europe Status and Prospect (2012-2022) 1.4.4 China Status and Prospect (2012-2022) 1.4.5 Japan Status and Prospect (2012-2022) 1.4.6 Southeast Asia Status and Prospect (2012-2022) 1.4.7 India Status and Prospect (2012-2022) 1.5 Global Market Size (Value) of High Performance Liquid Chromatography (HPLC) (2012-2022) 1.5.1 Global High Performance Liquid Chromatography (HPLC) Revenue Status and Outlook (2012-2022) 1.5.2 Global High Performance Liquid Chromatography (HPLC) Capacity, Production Status and Outlook (2012-2022)

2 Global High Performance Liquid Chromatography (HPLC) Market Competition by Manufacturers 2.1 Global High Performance Liquid Chromatography (HPLC) Capacity, Production and Share by Manufacturers (2012-2017) 2.1.1 Global High Performance Liquid Chromatography (HPLC) Capacity and Share by Manufacturers (2012-2017) 2.1.2 Global High Performance Liquid Chromatography (HPLC) Production and Share by Manufacturers (2012-2017) 2.2 Global High Performance Liquid Chromatography (HPLC) Revenue and Share by Manufacturers (2012-2017) 2.3 Global High Performance Liquid Chromatography (HPLC) Average Price by Manufacturers (2012-2017) 2.4 Manufacturers High Performance Liquid Chromatography (HPLC) Manufacturing Base Distribution, Sales Area and Product Type 2.5 High Performance Liquid Chromatography (HPLC) Market Competitive Situation and Trends 2.5.1 High Performance Liquid Chromatography (HPLC) Market Concentration Rate 2.5.2 High Performance Liquid Chromatography (HPLC) Market Share of Top 3 and Top 5 Manufacturers 2.5.3 Mergers & Acquisitions, Expansion

3 Global High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue (Value) by Region (2012-2017) 3.1 Global High Performance Liquid Chromatography (HPLC) Capacity and Market Share by Region (2012-2017) 3.2 Global High Performance Liquid Chromatography (HPLC) Production and Market Share by Region (2012-2017) 3.3 Global High Performance Liquid Chromatography (HPLC) Revenue (Value) and Market Share by Region (2012-2017) 3.4 Global High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue, Price and Gross Margin (2012-2017) 3.5 North America High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue, Price and Gross Margin (2012-2017) 3.6 Europe High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue, Price and Gross Margin (2012-2017) 3.7 China High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue, Price and Gross Margin (2012-2017) 3.8 Japan High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue, Price and Gross Margin (2012-2017) 3.9 Southeast Asia High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue, Price and Gross Margin (2012-2017) 3.10 India High Performance Liquid Chromatography (HPLC) Capacity, Production, Revenue, Price and Gross Margin (2012-2017)

4 Global High Performance Liquid Chromatography (HPLC) Supply (Production), Consumption, Export, Import by Region (2012-2017) 4.1 Global High Performance Liquid Chromatography (HPLC) Consumption by Region (2012-2017) 4.2 North America High Performance Liquid Chromatography (HPLC) Production, Consumption, Export, Import (2012-2017) 4.3 Europe High Performance Liquid Chromatography (HPLC) Production, Consumption, Export, Import (2012-2017) 4.4 China High Performance Liquid Chromatography (HPLC) Production, Consumption, Export, Import (2012-2017) 4.5 Japan High Performance Liquid Chromatography (HPLC) Production, Consumption, Export, Import (2012-2017) 4.6 Southeast Asia High Performance Liquid Chromatography (HPLC) Production, Consumption, Export, Import (2012-2017) 4.7 India High Performance Liquid Chromatography (HPLC) Production, Consumption, Export, Import (2012-2017)

Continued...

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[hitechtrader]

Dionex CD25 Conductivity Detector

Dionex Conductivity Detector CD25. Detects and quantifies ionic analytes in liquid and  ion chromatography. Range;0.01µS to    3000 uS, full scale.

[hitechtrader]

Dionex CD25 Conductivity Detector

Dionex Conductivity Detector CD25. Detects and quantifies ionic analytes in liquid and  ion chromatography. Range;0.01µS to    3000 uS, full scale.