MADAN SINGH’s Post

🔬Gas Chromatography: Your Ultimate Guide - Part I🔬 Gas Chromatography (GC) stands tall as a cornerstone technique in analytical chemistry, empowering scientists to dissect complex mixtures with precision. Let's delve deeper into its inner workings: 0️⃣ Inert Carrier Gas (Mobile Phase): Serving as the vehicle for sample transportation, the inert carrier gas propels the sample through the chromatographic column. (Helium, nitrogen, and hydrogen). 1️⃣ Gas Filters & Purifiers: Guaranteeing the purity of the carrier gas is paramount. Gas filters and purifiers scrub impurities from the gas stream, safeguarding against contamination. 2️⃣ Auto Samplers/Manual Syringes: Control the sample introduction into the GC system. Whether automated or manual, their precision and consistency in injecting samples directly impact the reliability of results. 3️⃣ Septum: Standing at the injection port, the septum forms a tight seal, preventing sample loss and contamination during injection. 4️⃣ Inlet Liners: protecting the chromatographic column from sample impurities, inlet liners aid in sample vaporization and minimize interactions with column surfaces. 5️⃣ Ferrules: Found at the column inlet and outlet, ferrules secure a leak-tight seal between the column and the inlet liner. 6️⃣ Column: It's where compounds are separated as they travel through, allowing for their identification and quantification. 7️⃣ Detector: At the end of the column, the detector captures separated sample components, converting them into measurable signals. (Flame Ionization Detector (FID) or Mass Spectrometer (MS)) 8️⃣ Chromatogram: The visual masterpiece of GC analysis, the chromatogram unveils the composition of the sample with striking clarity. Each peak represents a distinct compound, characterized by its retention time and peak area. Learn more at: https://lnkd.in/dGr2-45p. Send us your queries: inquiry@imeditec.com. #GasChromatography #gcms #gc #AnalyticalChemistry #Science #ChemicalAnalysis #Chromatography #Laboratory #Research #Chemistry #AnalyticalInstruments #Innovation #Technology #QualityControl #SeparationScience #ChemicalComposition #DataAnalysis #ResearchAndDevelopment #ScientificDiscovery #LabLife #STEM #ChemicalEngineering Trajan Scientific and Medical #innovativemeditech

🔬📊 HPLC Method Validation: A Step-by-Step Guide (Part 4) After explaining specificity, accuracy and precision in HPLC method validation we will continue in this post with Sensitivity (LOD & LOQ). The method detection limit (DL), also known as the limit of detection (LOD) is the smallest amount or concentration of analyte that can be detected. There are several ways to calculate LOD, as discussed in ICH Q2 (R1). The simplest way to calculate LOD is to determine the amount (or concentration) of an analyte that yields a peak height with a S/N of three. This can be performed for the materials of specified identified impurities that are available or a surrogate. During validation, the LOD should be verified with at least three injections of the diluted solution to meet at least three times the S/N. The quantitation limit (QL), also known as the limit of quantitation (LOQ), is the lowest level of an analyte that can be quantitated with some degree of certainty (with a precision of 5–10%). The simplest way to calculate LOQ is to determine the amount (or concentration) of an analyte that yields a peak with a S/N >10. Thus, LOQ is roughly equal to three times LOD. The LOQ must be less than or equal to the reporting threshold. It is customary to include a blank, sensitivity solution (by spiking the API at 0.05% or 0.1% level into the sample diluent), and a retention marker solution in the SST sequence to verify method sensitivity and specificity performance. There is another way to calculate LOD and LOQ by knowing the standard deviation of the response and the slope obtained from the calibration curve. you can find the way of calculations explained attached in this post. Share your chromatographic experiences below! 👇 💡📝

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🔬📊 HPLC Method Validation: A Step-by-Step Guide After we have discussed HPLC method development in our previous posts, now it is the time to validate our developed method. As analytical chemists, we understand the importance of robust and reliable methods. Whether you’re analyzing pharmaceuticals, environmental samples, or complex mixtures, method validation ensures that your HPLC results are accurate, precise, and reproducible. Let’s break down the validation process: 1. Linearity and Range: The Calibration Curve - Prepare a series of standard solutions covering the expected concentration range. Inject these standards into your HPLC system. - Plot peak area (y-axis) against concentration (x-axis). Ensure a linear relationship (R² ≥ 0.99) for accurate quantification. 2. Sensitivity: Detection Limit and Quantitation Limit - The method detection limit (DL), also known as the limit of detection (LOD) is the smallest amount or concentration of analyte that can be detected. - The quantitation limit (QL), also known as the limit of quantitation (LOQ), is the lowest level of an analyte that can be quantitated with some degree of certainty (with a precision of 5–10%) 3. Accuracy and Precision: The Twin Pillars - Accuracy: Compare your measured results to a known reference (e.g., certified standards). - Precision: Evaluate repeatability (intra-day) and intermediate precision (inter-day). Calculate %RSD (relative standard deviation) for peak areas and retention times. 4. Specificity and Selectivity: The Sherlock Holmes Test - Analyze your sample for potential interference from matrix components. - Use forced degradation studies (e.g., heat, acid, base) to assess stability-indicating power. 5. Robustness and System Suitability: Stress Testing - Vary method parameters (e.g., flow rate, column temperature) within acceptable limits. - Evaluate the impact on peak resolution, retention times, and peak areas. 6. Documentation: The Paper Trail - Document every step of your validation process. - Include acceptance criteria, deviations, and corrective actions. In our next posts we will take each item of validation procedure and discuss its importance and how to conduct them. Remember, method validation is like calibrating your analytical compass—essential for accurate navigation in the world of chromatography. Share your validation tips and experiences below! 👇💡🕺 #chromatography #hplc #researchanddevelopment #methoddevelopment #analyticalchemistry #methodvalidation #pharmaceuticalindustry #laboratory #iso17025 #method #metho #analyticalchemistry #hplcgc #gcms #lcms #kf #lcms

🔬📊 HPLC Method Validation: Understanding Specificity In the intricate world of high-performance liquid chromatography (HPLC), specificity is a critical validation parameter. It ensures that your method can accurately distinguish and quantify the analyte of interest in the presence of potential interferences. Let's delve into the specifics of specificity and how to assess it: 1. What Is Specificity? - Definition: Specificity refers to the ability of an HPLC method to selectively detect and quantify the target analyte without interference from other components in the sample matrix. - Importance: Specificity ensures that your method provides reliable results even in complex samples. 2. Conducting Specificity Studies: A Step-by-Step Approach 1. Selectivity Assessment: - Compare chromatograms of: - Blank: Mobile phase or sample matrix without the analyte. - Standard Solution: Analyte at a known concentration. - Sample Solution: Real sample containing the analyte. - Placebo Solution: Placebo is same as sample but without the desired analyte. - Look for any additional peaks in the sample solution that are absent in the blank and standard chromatograms. - If additional peaks are observed, investigate their origin (e.g., impurities, matrix components). 2. Forced Degradation Studies: - Subject the analyte to stress conditions (e.g., heat, acid, base, oxidation). - Analyze the degraded sample by HPLC. - Confirm that the analyte peak remains distinct and unaffected by degradation products. 3. Matrix Effects Evaluation: - Analyze the sample matrix without the analyte. - Spike the matrix with the analyte at a known concentration. - Compare peak areas between the spiked and unspiked samples. - Matrix effects should not significantly alter analyte quantification. 4. Specificity Acceptance Criteria: - The analyte peak should be well-resolved from other peaks. - No interference from impurities, degradation products, or matrix components. - Retention time and peak shape consistency across replicates. 3. Reporting Specificity Results: - Document the specificity study in your validation report. - Include chromatograms, peak purity assessments, and any relevant observations. Remember, specificity ensures that your HPLC method dances gracefully with precision, separating the analyte from the crowd. Share your chromatographic experiences below! 💃💡📝

Using gene expression profiling, our PM3CX test avoids challenges associated with immunostaining and biopsies of delicate tissue. Learn more about this unique test in this test-specific episode of our "Answers From the Lab" podcast. https://bit.ly/3U7Hc2L

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" Diving deep into the world of analytical chemistry, I embarked on a journey to construct a calibration curve using the timeless technique of manual injection. Armed with nothing but a syringe, vials, and a steady hand, each sample was meticulously introduced into the system, a testament to the artistry of scientific craftsmanship. 🌟 With each injection, I carefully observed, ensuring consistency and accuracy in every data point. No automation, no shortcuts - just the pure essence of hands-on experimentation. And the results speak volumes - an awe-inspiring R-squared value of 0.9995, a testament to the meticulous care and precision invested in the process. 💼 In a world dominated by automation, this experience served as a reminder of the power of human touch in scientific endeavors. It's not just about the data; it's about the journey, the dedication, and the sheer passion for discovery. Here's to manual mastery and the pursuit of scientific excellence! 💪 #ManualInjectionMastery #CalibrationCraftsmanship #ScientificExcellence"#thermofisherscientific