Inside O2H: Medicines, Molecules & More

Mobile Phase in HPLC – Types, Preparation & Role in Separation 💧 In High-Performance Liquid Chromatography (HPLC), the mobile phase is the liquid that moves through the column and carries the sample with it. Choosing the right mobile phase is critical for good separation, sharp peaks, and accurate results. In this post, you'll learn about the types of mobile phases used in HPLC, how to prepare them, and their impact on the analysis. What is the Mobile Phase in HPLC? The mobile phase is the solvent or mixture of solvents that flows through the HPLC column. It interacts with the stationary phase and the sample, helping separate the compounds based on polarity, charge, or size. Types of Mobile Phases 📌 Mobile phases are generally divided into two types: 1. Aqueous Phase (Water-Based) 💦 Usually includes water with a buffer (e.g., phosphate, acetate) Used in: Reverse-Phase HPLC pH control: Important for ionizable compounds 2. Organic Phase (Solvent-Base...

HPLC Detectors – Types and Their Applications in Pharma 🔍 After a sample is separated by the HPLC column, it needs to be detected and measured. This is where HPLC detectors come in. Detectors help us know what compound is coming out of the column and how much of it is present. Different detectors are used depending on the chemical nature of the compound. 1. UV-Visible Detector (UV/Vis & DAD) 🌈 This is the most commonly used detector in pharmaceutical analysis. It measures how much UV or visible light a compound absorbs. Single Wavelength UV: Measures absorbance at one fixed wavelength (e.g., 254 nm). DAD – Diode Array Detector: Records absorbance across a range of wavelengths. It helps in identifying peak purity and overlapping compounds. Best for: UV-absorbing compounds like drugs with aromatic rings or conjugated systems. Example: Paracetamol, aspirin, ibuprofen, antihistamines 2. Fluorescence Detector ✨ This detector measures the light emitted (fluor...

Understanding HPLC Columns: Types, Selection & Maintenance 🔬 The HPLC column is the most critical part of the chromatography system. It holds the stationary phase and is responsible for separating the compounds in your sample. The choice of column directly impacts the quality, speed, and reproducibility of your analysis. What is an HPLC Column? An HPLC column is a stainless steel tube packed with tiny particles called the stationary phase . When the liquid mobile phase pushes the sample through the column, interactions between the analytes and stationary phase cause them to separate and elute at different times. Types of HPLC Columns Columns are categorized based on the type of stationary phase they contain: Reverse-Phase Columns (RP): These are the most widely used columns in pharmaceutical analysis. They use non-polar stationary phases, commonly octadecyl (C18) or octyl (C8) bonded silica. Reverse-phase columns are excellent for separating non-polar to moderately ...

Types of HPLC – Explained with Examples 🔬 High-Performance Liquid Chromatography ( HPLC ) has different types depending on how the separation is done. Each type is designed to handle specific compounds and sample properties . In this post, you’ll learn the main types of HPLC used in the pharmaceutical industry and how they work. 1. Reverse-Phase HPLC (RP-HPLC) 🔄 Reverse-phase HPLC is the most commonly used type in pharma labs. In this method, the stationary phase is non-polar (hydrophobic), and the mobile phase is polar (water-based solvents). Used for: Non-polar or moderately polar compounds Example: Testing drug substances like paracetamol, ibuprofen, or vitamins 2. Normal-Phase HPLC (NP-HPLC) ⚖️ This method uses a polar stationary phase (like silica) and a non-polar mobile phase (like hexane). It separates polar compounds better than RP-HPLC. Used for: Separation of isomers, sugars, amino acids Example: Identification of sm...

Principle of HPLC – How It Works High-Performance Liquid Chromatography ( HPLC ) is a powerful analytical technique used in laboratories to separate mixtures into their individual chemical components . It helps scientists and pharmaceutical analysts understand the composition of complex samples by making each compound emerge one by one. What is the Stationary Phase? 🧱 Inside the HPLC system is a small tube called a column . This column is packed with tiny solid particles called the stationary phase . These particles remain fixed during analysis and provide the surface where chemical interactions happen. What is the Mobile Phase? 💧 The mobile phase is a liquid solvent or a mixture of solvents that flows continuously through the column. It acts as the carrier to push the sample components through the stationary phase, allowing the separation of compounds based on their chemical properties. How Does Separation Happen? 🔄 When the sample moves through the column:...

What is HPLC ? A Simple Introduction for Beginners High-Performance Liquid Chromatography (HPLC) is a powerful analytical technique used to separate, identify, and quantify the components in a mixture. It is widely applied in pharmaceutical, environmental, food, and chemical industries for quality control and research. In the pharmaceutical industry, HPLC is essential for ensuring the safety and efficacy of drugs by analyzing the purity, potency, and stability of active pharmaceutical ingredients (APIs) and formulations. Why is HPLC Important? Even minor variations in drug composition can lead to serious health risks. HPLC provides precise and reliable data that help manufacturers comply with regulatory standards and deliver safe medications to patients. It is also used for: Detecting impurities and degradation products Quantifying drug release rates in formulations Monitoring stability under different environmental conditions Basic Principle: HPLC separates c...