At BOC Sciences, we provide comprehensive 2D Chromatography services tailored to complex sample analysis across pharmaceuticals, biotechnology, fine chemicals, and natural products. Our expertise includes orthogonal separation using reversed-phase, HILIC, and ion-exchange columns, combined with multi-detector systems such as UV, DAD, fluorescence, and mass spectrometry. We design customized workflows to resolve overlapping peaks, map impurities, characterize protein isoforms, and profile metabolites with high resolution and reproducibility. Trusted by leading biopharmaceutical companies and research institutions, our solutions deliver rich, transparent, and reproducible data, helping clients accelerate development, optimize processes, and gain actionable analytical insights.
2D Chromatography is an advanced chromatographic technique that further develops traditional chromatography by combining two different separation mechanisms, typically orthogonal, to enhance separation power and data depth. This approach is particularly useful for analyzing complex samples, as it provides higher resolution and more comprehensive analytical information.
In 2D Chromatography, the sample is first separated on a primary column (first dimension) and then transferred to a secondary column (second dimension) for further separation. This two-dimensional separation allows components in complex mixtures to be resolved and identified more effectively. 2D Chromatography is widely applied across multiple fields, including pharmaceutical analysis, biochemistry, environmental science, and food science.
Our SEC×RP (Size Exclusion Chromatography × Reversed Phase) testing combines molecular size separation in the first dimension with hydrophobicity-based separation in the second dimension. This approach is highly effective for complex polymers and PROTAC impurities, allowing precise profiling of different molecular weight fractions and their corresponding chemical properties. The method ensures high-resolution separation, reliable quantitation, and reproducible analytical outcomes.
BOC Sciences' RP×RP testing employs two reversed-phase columns with pH modulation to achieve orthogonal separation. This technique is particularly suitable for small molecule pharmaceutical impurities where structural similarity can challenge single-dimension LC. Optimized gradient programs and pH switching allow clear differentiation of closely related compounds, delivering accurate and reproducible analytical results.
Our IEX×RP (Ion Exchange × Reversed Phase) testing targets proteins and biologics with charge variants or aggregation issues. The first dimension separates analytes based on charge, while the second dimension resolves hydrophobic differences. This workflow provides detailed characterization of protein heterogeneity, enabling confident interpretation of complex biotherapeutic profiles.
BOC Sciences' HILIC×RP (Hydrophilic Interaction Chromatography × Reversed Phase) workflow is optimized for polar metabolites and lipid analysis. The orthogonal separation ensures high-resolution profiling of compounds with diverse polarity, making it suitable for metabolomics studies, lipidomics, and complex natural product analysis.
Our NP×RP (Normal Phase × Reversed Phase) testing is designed for nonpolar mixtures such as oils, triglycerides, and their oxidation products. The first dimension separates based on polarity, followed by hydrophobic separation in the second dimension. This combination enables precise identification and quantification of minor components within complex lipid matrices.
BOC Sciences' GFC×RP (Gel Filtration × Reversed Phase) testing is tailored for antibody-drug conjugates (ADCs) and other large biomolecules. Gel filtration in the first dimension separates analytes by molecular size, while RP resolves hydrophobic differences, providing detailed DAR (drug-to-antibody ratio) analysis and thorough characterization of bioconjugates.
From unique separation challenges to specialized sample types, our scientists design 2D Chromatography methods tailored to your requirements. Request a quotation customized for your project.
Samples must dissolve completely in the first-dimension mobile phase. Avoid precipitation if solvent systems differ. Do not introduce non-volatile salts. Strong solvents like DMSO may be used only at ≤5 µL with proper dilution. For GC×GC, select low-boiling solvents such as hexane, ethyl acetate, or dichloromethane with minimal water.
Filter samples through 0.22 µm (LC) or 0.45 µm (GC) membranes; biological samples require protein precipitation and centrifugation before filtration. Suitable concentration ranges: 1–50 ppm for comprehensive 2D, or ≤2000 ppm for major components with trace ≥0.1 ppm. Preliminary 1D runs are recommended to confirm appropriate dilution and avoid column overloading.
Samples must be particle-free and stable. Light- or heat-sensitive materials should be stored at 4 ℃ in amber vials. For adsorption-prone biomolecules, add stabilizers such as Tween-20 or BSA. Injection volumes: 10–50 µL for comprehensive LC×LC, 100–1000 µL for heart-cutting, and ≤1 µL for GC×GC direct liquid injection.
Please provide your sample name, type, physical form, quantity, solubility, known impurities (if applicable), and any specific testing objectives or requirements. Our scientific team will carefully review your submission and design a tailored 2D Chromatography testing plan, along with a detailed quotation.
Raw data files include all one- and two-dimensional signals (UV, DAD, MS, etc.) and are delivered in open formats (.csv, .raw, .dx) or vendor-specific formats (.ch, .wiff), enabling future reprocessing or audit. Integration and quantitative tables provide 1D and 2D peak data with retention time, area/height, concentration, recovery, and RSD. Visualizations include 1D chromatograms, 2D plots, contour or 3D waterfall charts with labeled peaks, cut regions, and spectral extraction points for intuitive review of separation quality.
Reports include spectral purity and impurity assessment with UV match factors and threshold evaluation, indicating whether each peak is spectrally pure. Full method parameters (gradient, valve timing, flow rate, temperature) and system suitability test results, such as theoretical plates, tailing factor, resolution, and signal-to-noise ratio, are documented. Instrument diagnostics and EMF (Early Maintenance Feedback) counts are also provided to ensure traceable and reliable data for your analysis requirements.
Optional advanced analysis can include outputs from software such as GC Image or MassHunter, featuring PCA, Volcano plots, peak capacity, and 2D resolution metrics. Multi-batch statistical evaluation, confidence intervals, and significance testing are also supported, enabling comprehensive insights for complex studies, metabolomics, or other high-resolution, multi-component analyses. Custom calculations can be tailored to meet project-specific research objectives and analytical goals.
Pharmaceutical companies require high-resolution separation of drug substances, impurities, and degradation products. Our 2D Chromatography testing provides detailed chromatograms, peak integration, and spectral purity analysis. Multi-dimensional separation enables comprehensive profiling of complex formulations, supporting formulation optimization, stability studies, and quality assessment with reliable, traceable results for R&D and analytical decision-making.
Biotech scientists working with proteins, peptides, and biologics need precise separation of heterogeneous biomolecules. Our 2D Chromatography services deliver high-resolution mapping of protein isoforms, post-translational modifications, and aggregation states. Detailed chromatograms, purity evaluation, and advanced statistical analysis facilitate biotherapeutic development, process optimization, and batch-to-batch comparability.
Developers of specialty chemicals and multi-component materials face challenges in analyzing complex mixtures. Our 2D Chromatography solutions offer comprehensive separation, impurity profiling, and quantitative analysis of target and by-product components. High-resolution data and traceable reports support formulation development, process optimization, and quality monitoring of advanced materials.
Researchers in natural products, metabolomics, and complex biological extracts require detailed separation of multiple components with overlapping properties. Our 2D Chromatography testing provides high-resolution chromatograms, peak identification, and quantitative data, enabling compound profiling, metabolite mapping, and comparative studies across samples with actionable insights for research and discovery.
The client completes the sample information form, specifying matrix type, target analytes, concentration range, and special requirements. The lab evaluates separation difficulty, selecting 2D mode, estimated injections, and detector configuration (UV/DAD/MS).
Clients send ≥2 samples (1 primary, 1 backup). The lab conducts 1D screening to locate target windows and interference peaks, then designs the 2D valve event table and completes system suitability testing (RSD ≤2%, theoretical plates and tailing factors within specifications).
Samples are sequenced: blank → system suitability → client samples in duplicate → spiked recovery → QC. The 2D-LC system automatically performs 1D separation, valve switching, 2D separation, detector acquisition, and real-time pressure monitoring.
Software splits 2D slices into individual chromatograms. Peaks are identified with ±0.2 min retention time tolerance and spectral matching factor ≥990. Quantitative calculations provide content, recovery, and RSD statistics.
Analysis and review are completed, producing the final report including 1D/2D chromatograms, contour plots, method parameters, and system suitability results (SST triple injection), ensuring data completeness and traceability.
Raw data are securely stored, and clients may request reprocessing or data review. Technical support is provided, including adjustment of integration parameters and statistical recalculation.
Many leading biopharmaceutical companies choose to collaborate with us, relying on our 2D chromatography expertise for high-resolution analysis and reliable, professional data support.
We combine UV, DAD, fluorescence, and mass spectrometry detectors to provide complementary qualitative, quantitative, and structural information, delivering more comprehensive insights from a single workflow.
Our services support simultaneous analysis of multiple samples and offer multi-sample discounts, reducing costs while improving efficiency for high-throughput research and development.
Complete raw and processed data are provided, including peak integration and method parameters, ensuring traceable, reproducible results that support research and process decision-making.
Access essential chromatography resources, including guidance on column selection, mobile phase optimization, method setup, data interpretation, and detector usage tips.
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Client: A leading phytochemical research institute in North America
Challenge: The client needed to separate and quantify structurally similar bioactive compounds from complex plant extracts, where traditional 1D liquid chromatography showed co-elution issues.
Solution: BOC Sciences implemented a two-dimensional liquid chromatography workflow, combining reversed-phase and hydrophilic interaction chromatography (HILIC) columns. Our experts optimized modulation times, gradient programs, and detection methods to achieve high-resolution separation. Detailed chromatograms and quantitative reports were provided, enabling precise identification of minor components.
Outcome: The client achieved superior compound resolution, reliable quantification of trace components, and a streamlined workflow that reduced analysis time by 30%, improving overall laboratory efficiency.
Client: A global flavor and fragrance company
Challenge: The client required comprehensive profiling of volatile compounds in essential oils and fragrances, including trace-level components that were challenging to detect with conventional GC.
Solution: BOC Sciences deployed comprehensive two-dimensional gas chromatography (GC×GC) with dual flame ionization and time-of-flight mass spectrometry detection. Column combinations and modulation parameters were carefully optimized to maximize peak capacity and separation of closely eluting compounds. Our team delivered detailed contour plots and component tables for rapid interpretation.
Outcome: The client obtained a complete chemical profile of their products, enabling enhanced quality control, accurate batch-to-batch comparison, and confident decision-making. Analytical throughput improved without compromising resolution.
Client: A mid-size pharmaceutical manufacturer in Asia
Challenge: The client faced challenges in identifying and quantifying low-abundance impurities in complex drug formulations, where single-dimensional LC-MS/MS failed to provide sufficient separation.
Solution: BOC Sciences implemented 2D LC-MS/MS, combining reversed-phase and ion-exchange chromatography. Optimized multi-dimensional separation strategies, including orthogonal mobile phase selection and gradient programming, were applied to enhance resolution. Our team provided comprehensive impurity profiling with quantitative reports aligned with the client's internal standards.
Outcome: The client achieved accurate impurity identification at trace levels, improved process monitoring, and faster decision-making for formulation development. Analytical reliability and throughput were significantly enhanced, supporting product quality assurance.
2D chromatography is particularly effective for complex matrices where one-dimensional separation fails to resolve co-eluting compounds. Typical samples include natural product extracts, multi-component formulations, volatile mixtures, and pharmaceutical impurities. The orthogonal separation strategies enable high-resolution analysis of trace-level components that would otherwise remain undetected.
Proper sample preparation is critical to ensure reproducibility and accurate detection. Factors such as solvent selection, filtration, and sample concentration must be carefully optimized to avoid column overload, peak broadening, or matrix interference. For sensitive analyses, pre-fractionation or cleanup steps may be recommended to maximize resolution and signal-to-noise ratio.
Key challenges include selecting complementary columns for orthogonal separation, optimizing modulation or transfer times between dimensions, and fine-tuning gradient programs. Insufficient orthogonality or incorrect modulation parameters can lead to peak co-elution or reduced peak capacity. Our team routinely addresses these issues with methodical optimization to achieve reproducible high-resolution separations.
Detection sensitivity directly influences the ability to identify low-abundance or trace-level components. In 2D chromatography, peak dilution across dimensions can reduce signal intensity. High-sensitivity detectors, appropriate injection techniques, and careful method optimization are essential to maintain quantifiable signals while achieving maximal resolution.
Interpreting 2D chromatograms requires understanding both dimensions' retention behavior and the correlation between peaks. Contour plots or 3D representations are commonly used to visualize complex separations. Accurate integration, peak assignment, and reproducibility checks are critical for reliable quantitative analysis and for drawing meaningful conclusions from complex sample matrices.
The 2D chromatography service from BOC Sciences provided outstanding separation of complex mixtures that were previously difficult to resolve. Their expertise in method optimization and column selection allowed us to clearly identify minor components with confidence.
— Dr. S., Senior Scientist, Natural Products Research Institute
BOC Sciences' 2D LC-MS/MS testing consistently delivers comprehensive analytical reports, including high-resolution chromatograms and quantitative data. Their team's attention to detail and clear documentation have greatly enhanced our research workflow.
— Dr. M., Head of Analytical Chemistry, Pharmaceutical R&D Lab
Working with BOC Sciences on 2D GC×GC for volatile compound profiling has significantly improved our ability to analyze complex fragrance and flavor mixtures. The service's efficiency and accuracy allowed us to optimize product formulations more effectively.
— Ms. R., Product Development Manager, Global Flavor & Fragrance Company
The team at BOC Sciences provided expert guidance throughout our 2D chromatography projects. Their collaborative approach and technical insights ensured method development and data interpretation were smooth, helping us make informed research decisions.
— Dr. T., Principal Scientist, Biotech Analytical Department
