Liquid Chromatography Mass Spectrometry (LC-MS) Analysis

Liquid Chromatography-Mass Spectrometry (LC-MS) is a powerful analytical technique widely utilized at Emery Pharma for its exceptional sensitivity, selectivity, and versatility in analyzing complex samples within the pharmaceutical outsourcing and drug development pipeline. As a leading Contract Research Organization (CRO), we leverage LC-MS to provide clients with reliable data and accurate results crucial for regulatory submission and product development.

Types of Analyses Performed Using LC-MS

LC-MS integrates the separation capabilities of liquid chromatography (HPLC or UPLC) with the detection and identification power of mass spectrometry. This combination allows for both qualitative characterization and quantitative analysis across various matrices.

1. Qualitative Analysis (Characterization and Identification):
LC-MS, especially with High-Resolution Mass Spectrometry (HRMS) capabilities, is indispensable for:

• Structural Elucidation: Identifying unknown compounds, impurities, degradation products, and metabolites in drug substances and drug products. This is critical for understanding molecular structures.
• Impurity Profiling: Comprehensive identification of process-related impurities and degradation impurities, including potentially harmful nitrosamine impurities (NDSRI testing), to ensure product safety and FDA compliance.
• Biologic Characterization: Analyzing and characterizing complex biologics such as peptides, proteins, and Antibody-Drug Conjugates (ADCs), including their modifications and variants.
• Formulation Component Analysis: Identifying and characterizing excipients and other components, such as the quantitation and characterization of Polysorbate 80 or Poloxamers in various products.
• Biomarker Discovery: Identifying and confirming potential biomarkers in biological samples for disease research or therapeutic monitoring.

2. Quantitative Analysis (Quantitation):
LC-MS offers superior sensitivity and specificity for precise quantitation of target analytes. Key quantitative applications include:

• Bioanalytical Services: High-throughput quantitation of drugs and their metabolites in various biological matrices (plasma, urine, tissues) for pharmacokinetic (PK) and pharmacodynamic (PD) studies, supporting preclinical and clinical trials. This is a core part of the drug development pipeline.
• Impurity Quantitation: Accurate determination of the levels of identified impurities in accordance with ICH guidelines and cGMP/GLP standards for lot release testing and stability studies.
• Active Pharmaceutical Ingredient (API) Quantitation: Measuring the concentration of active pharmaceutical ingredients in drug products to ensure proper dosage and quality.
• Stability Studies: Monitoring drug degradation over time by precisely quantifying drug substance and its degradation products under various stress conditions.
• Dissolution Testing: Quantifying drug release from formulations over time.
• Antimicrobial Studies Support: While LC-MS doesn't directly perform MIC/MBC, it can quantify drug concentrations in media or biological samples to support antimicrobial effectiveness testing (AET) or other related studies.

At Emery Pharma, our scientific expertise includes robust Method Development and Method Validation for these applications, ensuring that the analytical methods are fit-for-purpose and compliant with all relevant regulatory requirements.

Detection Limits of LC-MS

Key Factors Influencing Detection Limits:

• Analyte Properties: The inherent chemical properties of the compound, such as its ionization efficiency in the mass spectrometer, significantly impact its detectability.
• Matrix Effects: The complexity of the sample matrix (e.g., biological fluids, highly formulated products) can interfere with ionization or signal suppression, affecting sensitivity.
• LC-MS System Configuration: The type of mass spectrometer (e.g., triple quadrupole for high sensitivity quantitation vs. HRMS for accurate mass and structural information) and the specific LC conditions (column, flow rate, mobile phase) are optimized during method development.
• Method Optimization: Thorough method development and validation are crucial for achieving the lowest possible detection and quantitation limits for a given analyte and matrix. Our scientific expertise ensures optimal conditions are met for your specific project scope.

Our project management team works closely with clients to understand their specific analytical needs, including required sensitivity for FDA compliance and overall drug development goals. We strive to achieve the lowest possible detection limits through tailored solutions, providing the accurate results and reliable data necessary for informed decision-making and successful regulatory submissions. For specific project quotes or to discuss rush analysis capabilities, please contact us to leverage our scientific expertise.

Emery Pharma specializes in providing robust LC-MS/MS bioanalytical services that are fundamental for pharmacokinetics (PK) and drug metabolism (ADME) studies. Our comprehensive support helps pharmaceutical and biotechnology companies generate the reliable data needed to advance their drug candidates through the drug development pipeline and meet FDA compliance requirements for regulatory submission.

Here’s how Emery Pharma supports your LC-MS/MS assay needs:

The Critical Role of LC-MS/MS in PK/ADME Studies

Pharmacokinetics (PK) studies are essential for understanding how a drug is absorbed, distributed, metabolized, and excreted by the body. Drug metabolism studies identify and quantify the metabolites formed from the parent drug. LC-MS/MS (Liquid Chromatography-Tandem Mass Spectrometry) is the gold standard for these studies due to its high sensitivity, selectivity, and versatility in quantifying drug compounds and their metabolites at trace levels within complex biological matrices (e.g., plasma, urine, tissues).

Our Comprehensive LC-MS/MS Bioanalytical Services

Emery Pharma leverages extensive scientific expertise and state-of-the-art instrumentation to deliver precise and accurate results for your bioanalytical projects.

1. Expertise & Instrumentation:

• We utilize advanced LC-MS/MS platforms, often coupled with UPLC (Ultra-Performance Liquid Chromatography) or HPLC, to achieve superior chromatographic separation and detection limits.
• Our capabilities extend to High-Resolution Mass Spectrometry (HRMS) for in-depth characterization and structural elucidation of metabolites, providing critical insights into drug metabolism pathways.

2. Method Development & Validation:

• Method Development: Our scientists design and optimize sensitive and selective LC-MS/MS methods tailored to your specific drug candidate, biological matrix, and analytical requirements. This involves optimizing sample preparation, chromatographic separation, and mass spectrometric parameters to ensure robustness and minimize matrix effects.
• Method Validation: We perform rigorous Method Validation in accordance with GLP (Good Laboratory Practice) principles and global ICH guidelines. This process thoroughly evaluates critical assay parameters such as accuracy, precision, linearity, range, sensitivity (LLOQ/ULOQ), and stability, ensuring the method's fitness for purpose and generating FDA-compliant data.

3. Sample Analysis & Reporting:

• With fully validated methods, we conduct high-throughput analysis of preclinical and clinical study samples. Our strict quality control procedures throughout the entire process ensure the integrity and reproducibility of all analytical data.
• We provide comprehensive and transparent reports that include all raw data, processed results, summary tables, and interpretations, facilitating your data review and subsequent regulatory submission.

Ensuring Quality and Client-Focused Partnership

As your trusted CRO, we are committed to providing exceptional service and supporting your pharmaceutical outsourcing needs.

• GLP Compliance: All bioanalytical activities are conducted under strict GLP regulations, ensuring the integrity and traceability of your data for regulatory purposes.
• Project Management: Our dedicated management team ensures clear communication, transparent project scope, and efficient execution from initial quote to final report.
• Turnaround Time (TAT): We understand the importance of timely data in drug development. We work diligently to optimize Turnaround Time (TAT) and can discuss options for rush analysis to meet your most critical timelines, without compromising on quality.

By partnering with Emery Pharma for your LC-MS/MS bioanalytical services, you gain a scientific ally dedicated to providing the reliable data and accurate results essential for successful drug development.

Gas Chromatography-Mass Spectrometry (GC-MS) is a powerful analytical technique ideally suited for the separation, identification, and quantitation of a wide range of volatile compounds and semi-volatile compounds. The suitability of a compound for GC-MS primarily depends on its thermal stability, volatility, and molecular weight.

Characteristics of Suitable Compounds:

Compounds best analyzed by GC-MS typically possess:

• Volatility: They must readily vaporize when heated without decomposing.
• Thermal Stability: They must remain chemically intact at the elevated temperatures required for the GC separation (typically up to 300-350°C).
• Lower to Moderate Molecular Weight: Generally, compounds with molecular weights up to approximately 600-800 Da are good candidates.

Key Applications of GC-MS in Pharmaceutical and Biotech Industries

At Emery Pharma, our scientific expertise in GC-MS plays a critical role in various stages of the drug development pipeline, ensuring reliable data and accurate results for our clients. Common applications include:

1. Residual Solvent Analysis: GC-MS is the gold standard for detecting and quantifying residual solvents (e.g., ethanol, methanol, acetone, toluene, dichloromethane) in pharmaceutical excipients, active pharmaceutical ingredients (APIs), and finished drug products, adhering to ICH guidelines and FDA compliance. This is a routine part of lot release testing.
2. Impurity Profiling and Degradation Product Analysis: We use GC-MS for identifying and quantifying volatile impurities and degradation products that may arise during synthesis, manufacturing, or storage of drug substances and products. This is often integrated into stability studies.
3. Extractables and Leachables (E&L) Testing: For drug packaging, medical devices, and manufacturing components, GC-MS is essential for identifying and quantifying volatile and semi-volatile extractables and leachables that could migrate into the drug product, affecting its safety or efficacy.
4. Raw Material Testing: Ensuring the quality and purity of incoming raw materials by detecting and quantifying volatile contaminants.
5. NDSRI Testing (Nitrosamine Drug-Substance Related Impurities): While many nitrosamines are analyzed by LC-MS or High-Resolution Mass Spectrometry (HRMS), certain volatile nitrosamines can be effectively characterized and quantified using GC-MS as part of comprehensive NDSRI testing.
6. Characterization of Volatile Components: From flavor and fragrance compounds in consumer health products to off-gassing from materials, GC-MS provides comprehensive characterization.

Emery Pharma's Comprehensive GC-MS Services:

As a leading Contract Research Organization (CRO), Emery Pharma offers end-to-end support for your analytical needs, leveraging our robust project management system. Our GC-MS capabilities include:

• Method Development: Our chemists specialize in developing customized and sensitive GC-MS methods for unique or challenging matrices, tailored to your specific project scope.
• Method Validation: We rigorously validate all methods according to cGMP and GLP standards, ensuring they are suitable for their intended purpose and meet regulatory submission requirements.
• Structural Elucidation: In conjunction with other techniques like NMR analysis (including qNMR) and HRMS, GC-MS aids in the structural elucidation of unknown volatile compounds.
• Quality and Turnaround Time (TAT): We are committed to delivering high-quality, accurate results with efficient Turnaround Time (TAT), including options for rush analysis when critical. We provide transparent quotes and detailed reports.

For compounds that are thermally labile, non-volatile, or have high molecular weights, Emery Pharma offers a full suite of complementary analytical techniques, including HPLC, UPLC, LC-MS, High-Resolution Mass Spectrometry (HRMS), and NMR analysis, ensuring we can meet all your characterization and bioanalytical services needs for pharmaceutical outsourcing.

At Emery Pharma, a leading Contract Research Organization (CRO), we adhere strictly to established international and national regulatory guidelines to ensure the robustness, reliability, and regulatory compliance of all our LC-MS Method Validation activities. Our commitment is to provide accurate results and reliable data essential for your drug development pipeline and successful regulatory submission.

Our comprehensive approach to LC-MS Method Validation is primarily guided by:

Key Regulatory Guidelines

1. International Council for Harmonisation (ICH) Guidelines:

• Specifically, ICH Q2(R2) "Validation of Analytical Procedures: Text and Methodology" serves as a foundational document for our Method Validation processes. This guideline provides a framework for evaluating parameters such as specificity, linearity, range, accuracy, precision, detection limit (LOD), quantitation limit (LOQ), and robustness.
• While ICH Q2(R2) is broad, we apply its principles meticulously to the unique demands of LC-MS applications, including those involving High-Resolution Mass Spectrometry (HRMS) for complex characterization and quantitation.

2. U.S. Food and Drug Administration (FDA) Guidance for Industry:

• We closely follow relevant FDA guidance documents, particularly the "Guidance for Industry: Bioanalytical Method Validation." This guidance is critical for studies supporting pharmacokinetic (PK), pharmacodynamic (PD), and toxicokinetic (TK) evaluations, ensuring our bioanalytical services meet the stringent requirements for clinical and non-clinical trial data used in regulatory submissions.

3. Good Laboratory Practice (GLP) and Good Manufacturing Practice (cGMP):

• Our Method Development and Method Validation activities are conducted within a quality system that incorporates principles of GLP and cGMP, as applicable to the specific project scope. This ensures the integrity, traceability, and reproducibility of data generated for studies ranging from early-stage research to stability studies and lot release testing. Adherence to these quality systems underscores our dedication to providing quality data throughout the pharmaceutical outsourcing process.

Our LC-MS Method Validation Process

Our validation process systematically evaluates the following key parameters to ensure the analytical method is fit for its intended purpose, delivering the scientific expertise and reliable data you need:

• Specificity/Selectivity: Demonstrating the ability of the LC-MS method to unequivocally assess the analyte in the presence of components that may be expected to be present (e.g., impurities, degradation products, matrix components).
• Sensitivity: Determining the Limit of Detection (LOD) and Limit of Quantitation (LOQ) to ensure the method can accurately measure the analyte at very low concentrations.
• Accuracy: Assessing the closeness of test results to the true value.
• Precision: Evaluating the degree of agreement among individual test results when the procedure is applied repeatedly to homogeneous samples (repeatability and intermediate precision).
• Linearity and Range: Establishing the range over which the analytical method provides results that are directly proportional to the concentration of the analyte.
• Recovery: For bioanalytical services, this includes assessing the efficiency of analyte extraction from complex biological matrices.
• Stability: Investigating the stability of the analyte in solutions and within the sample matrix under various storage and processing conditions, crucial for stability studies and ensuring accurate results over time.

Our project management team works closely with clients to define the project scope, ensuring that all LC-MS Method Validation efforts are tailored to their specific needs and regulatory pathway, aiming for optimal Turnaround Time (TAT) without compromising quality.

How LC-MS Helps in Identifying and Quantifying Drug-Related Impurities

Liquid Chromatography-Mass Spectrometry (LC-MS) is an indispensable analytical technique in the pharmaceutical industry, playing a critical role in the identification and quantitation of drug-related impurities. As a Contract Research Organization (CRO), Emery Pharma leverages advanced LC-MS platforms, including High-Resolution Mass Spectrometry (HRMS), to ensure the safety, quality, and efficacy of drug products throughout the drug development pipeline.

Why Impurity Analysis is Critical

Drug-related impurities, even at trace levels, can impact drug safety and effectiveness. Regulatory bodies like the FDA and global ICH guidelines mandate strict control over impurities. Our scientific expertise ensures thorough characterization and accurate quantitation to support FDA compliance and successful regulatory submission. This includes vital NDSRI testing to address recent regulatory requirements for nitrosamine impurities.

The LC-MS Process for Impurity Identification

LC-MS combines the powerful separation capabilities of Liquid Chromatography (HPLC or UPLC) with the highly sensitive and selective detection of Mass Spectrometry.

1. Chromatographic Separation (LC): The sample is first introduced into the LC system, which separates components based on their physicochemical properties. This isolates individual impurities from the active pharmaceutical ingredient (API) and other excipients, allowing for their distinct analysis.

2. Mass Spectrometric Detection (MS): Eluting compounds from the LC are then introduced into the mass spectrometer.

• Ionization: Molecules are ionized, converting them into charged particles.
• Mass Analysis: These ions are directed through a mass analyzer, which separates them based on their mass-to-charge ratio (m/z). This provides a unique "fingerprint" for each compound.
• High-Resolution Mass Spectrometry (HRMS): With HRMS, we obtain highly accurate mass measurements, enabling the determination of elemental composition and differentiation of compounds with very similar nominal masses.

3. Structural Elucidation: For unknown impurities, fragmentation patterns obtained through tandem MS (MS/MS) provide crucial information about their molecular structure. This data, combined with our scientific expertise and often supplemented by NMR analysis for complete structural elucidation, allows us to definitively identify the impurity.

The LC-MS Process for Impurity Quantitation

Once identified, LC-MS is also exceptionally powerful for quantitation, even at very low levels:

1. Specific Ion Monitoring: The mass spectrometer can be set to specifically monitor the unique mass-to-charge ratio of an identified impurity. This selective detection significantly reduces interference and increases sensitivity.
2. Calibration Curves: Known concentrations of impurities are analyzed to generate calibration curves. By comparing the signal intensity of the impurity in the sample to these curves, its exact concentration can be quantified.
3. Trace Level Detection: The high sensitivity of modern LC-MS systems, especially HRMS, allows for the accurate results and reliable data needed to detect and quantify impurities present at extremely low, sub-ppm levels, critical for ICH guidelines compliance.

Emery Pharma's Expertise in Impurity Analysis

At Emery Pharma, our comprehensive bioanalytical services encompass robust Method Development and rigorous Method Validation for impurity profiling and quantitation, adhering to cGMP and GLP standards. We specialize in:

• Stability Studies: Monitoring impurities over time under various conditions to understand degradation pathways.
• Lot Release Testing: Ensuring that each batch of drug product meets specified impurity limits before release.
• NDSRI Testing: Our specialized analytical methods are designed to identify and quantify complex nitrosamine impurities in drug substances and products, supporting clients in meeting the August 2025 FDA deadline.

Our dedicated project management team ensures efficient communication, clear project scope definition, and timely delivery of reliable data. We understand the importance of Turnaround Time (TAT) and are equipped to handle diverse analytical needs, from initial quote requests to rush analysis when required, making us a preferred partner for pharmaceutical outsourcing.

Yes, Emery Pharma routinely performs residual solvent testing using Headspace Gas Chromatography-Mass Spectrometry (HS-GC-MS) in strict adherence to USP <467> guidelines. This crucial analysis ensures the safety and quality of pharmaceutical products by detecting and quantifying volatile organic impurities that may remain from the manufacturing process.

Understanding USP <467> and Residual Solvents

Residual solvents are organic volatile chemicals used or produced in the manufacture of drug substances, excipients, or drug products. While some solvents are unavoidable, their levels must be carefully controlled to meet regulatory limits for patient safety. USP <467> (Residual Solvents) provides a comprehensive framework, classifying solvents into three classes based on their toxicity and outlining acceptable daily exposures (ADEs) and analytical methods. Our analyses comply with these requirements, aligning with ICH Q3C (R9) guidelines for residual solvents.

Our Expert Approach to Residual Solvent Analysis

At Emery Pharma, our scientific expertise and state-of-the-art laboratory are equipped to handle a wide array of matrices for residual solvent detection.

• Headspace GC-MS Technology: We leverage advanced Headspace Gas Chromatography-Mass Spectrometry (HS-GC-MS), which is the gold standard for this analysis. This technique efficiently extracts volatile compounds from samples and separates them for precise quantitation and characterization, providing accurate results.
• Regulatory Compliance: All testing is performed under cGMP (current Good Manufacturing Practices) and GLP (Good Laboratory Practices) compliant conditions, ensuring the data generated is suitable for FDA compliance and regulatory submissions.
• Method Development and Validation: For unique sample matrices or specific client requirements, we offer Method Development and Method Validation services tailored to your project scope. This ensures the analytical method is robust, reliable, and fit for its intended purpose, critical for lot release testing and throughout the drug development pipeline.

Project Workflow and Turnaround Time (TAT)

1. Initial Consultation & Quote: We begin with a detailed discussion of your specific testing requirements. A comprehensive quote and proposed timeline will be provided.
2. Sample Submission: Clients submit samples, along with any relevant documentation (e.g., Certificates of Analysis for starting materials, known solvents used).
3. Analysis Execution: Our experienced scientists perform the residual solvent analysis using validated HS-GC-MS methods.
4. Data Interpretation & Reporting: The generated data undergoes rigorous review by our experts, followed by the issuance of a detailed report, including all detected solvents, their concentrations, and compliance against USP <467> limits.

Turnaround Time (TAT):
Standard Turnaround Time (TAT) for routine residual solvent testing typically ranges from X to Y business days, though this can vary.

• Factors influencing TAT: The complexity of the sample matrix, the need for new Method Development or Method Validation, the number of samples, and current laboratory workload can all impact the timeline.
• Rush Analysis: For urgent needs, we offer rush analysis options to expedite your project. Please discuss your specific timelines with our project management team during the initial consultation.

Your Trusted CRO Partner

As a leading Contract Research Organization (CRO) specializing in pharmaceutical outsourcing, Emery Pharma is committed to providing comprehensive analytical services that support your entire drug development pipeline. Our dedication to quality, scientific expertise, and client-focused project management ensures you receive reliable data and accurate results critical for product safety and regulatory success.