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Chromatography | Interactive Continuous Simulation | Paper Chromatography

Chromatography: Principles, Methods

Separation technique — stationary phase, mobile phase, retention factor, types of chromatography

🧪 Real-time Paper Chromatography

Solvent rises continuously. Components separate by different Rf values. Simulation loops automatically.

Red / Fast Yellow Blue / Slow
Solvent front rising — components separating by Rf
Water (polar) Ethanol (moderate) Hexane (non-polar)
Dye mixture Marker inks

Chromatography is a powerful separation technique used to separate, identify, and purify components of a mixture. It exploits differences in how substances partition between a stationary phase (solid or liquid) and a mobile phase (liquid or gas). Components migrate at different rates, allowing separation. It is fundamental in chemistry, biology, forensics, environmental science, and pharmaceutical analysis.

Principle of Chromatography

The mixture is dissolved in the mobile phase, which carries it through the stationary phase. Each component has a different affinity for the two phases → different migration speeds → separation. The retention factor (Rf) is a key parameter:

Rf = distance travelled by compound / distance travelled by solvent front

Rf is characteristic for a given compound under fixed conditions. It helps in identification.

Types of Chromatography

📄 Paper Chromatography

Stationary phase: paper (cellulose). Mobile phase: solvent. Used for separating dyes, amino acids, simple mixtures.

🧪 Thin Layer Chromatography (TLC)

Stationary phase: silica gel or alumina coated on glass plate. Fast, high resolution, widely used in organic chemistry.

🔬 Column Chromatography

Stationary phase packed in a vertical column. Mobile phase flows through, fractions collected. Used for preparative separations.

💨 Gas Chromatography (GC)

Mobile phase: inert gas (He, N₂). Stationary phase: liquid coated on solid support. For volatile compounds.

💧 High Performance Liquid Chromatography (HPLC)

High pressure forces mobile phase through column. High resolution, quantitative analysis.

🧬 Ion Exchange Chromatography

Separates ions and polar molecules based on charge. Used in protein purification, water softening.

Retention Factor (Rf) and Identification

After development, the distances are measured. Rf values between 0 and 1 indicate separation. Unknown compounds can be identified by comparing Rf with standards.

Example: Rf (blue dye) = 3.2 cm / 6.0 cm = 0.53

Applications of Chromatography

  • Pharmaceuticals: Purity testing, drug analysis, metabolite detection.
  • Forensics: Analysis of ink, dyes, drugs, and toxicological samples.
  • Environmental: Monitoring pesticides, pollutants, water contaminants.
  • Food industry: Quality control, preservatives, vitamin analysis.
  • Biochemistry: Protein purification, DNA sequencing (HPLC, GC).

📝 Chromatography – Quiz

1. What is the stationary phase in paper chromatography?

2. The formula for retention factor (Rf) is:

3. Which type of chromatography uses an inert gas as mobile phase?

4. In TLC, if a compound travels 4.2 cm and solvent front travels 6.0 cm, Rf is:

5. Which technique is best for separating heat-sensitive, non-volatile compounds?

🎥 Complete Lecture: Chromatography (Urdu/Hindi)
📥 Download Complete Notes (PDF)

Click to download comprehensive notes on chromatography.

© 2025 — Complete guide to Chromatography with continuous simulation. Principle, methods, interactive simulation, quiz, and video lecture.

(function () { var canvas = document.getElementById(‘chromaCanvas’); var ctx = canvas.getContext(‘2d’); var solventSelect = document.getElementById(‘solventSelect’); var mixtureSelect = document.getElementById(‘mixtureSelect’); var speedSlider = document.getElementById(‘speedSlider’); var resultBox = document.getElementById(‘chromaResult’); var resetBtn = document.getElementById(‘resetBtn’); var pauseBtn = document.getElementById(‘pauseBtn’); var playBtn = document.getElementById(‘playBtn’);var W = 520, H = 400; var PAPER_L = 90, PAPER_R = 430; var PAPER_TOP = 30, PAPER_BOT = 370; var BASELINE = 330; var SOLVENT_START = 355;var spots = []; var solventY = SOLVENT_START; var running = true; var last = 0; var simTime = 0; var loopDelay = 0;function getRfData() { var solvent = solventSelect.value; var mixture = mixtureSelect.value; var data = [];if (mixture === ‘dyes’) { if (solvent === ‘water’) { data = [ { name: ‘Red’, color: ‘#e74c3c’, rf: 0.72 }, { name: ‘Yellow’, color: ‘#f1c40f’, rf: 0.88 }, { name: ‘Blue’, color: ‘#2980b9’, rf: 0.42 } ]; } else if (solvent === ‘ethanol’) { data = [ { name: ‘Red’, color: ‘#e74c3c’, rf: 0.55 }, { name: ‘Yellow’, color: ‘#f1c40f’, rf: 0.75 }, { name: ‘Blue’, color: ‘#2980b9’, rf: 0.38 } ]; } else { data = [ { name: ‘Red’, color: ‘#e74c3c’, rf: 0.22 }, { name: ‘Yellow’, color: ‘#f1c40f’, rf: 0.30 }, { name: ‘Blue’, color: ‘#2980b9’, rf: 0.12 } ]; } } else { if (solvent === ‘water’) { data = [ { name: ‘Cyan’, color: ‘#1abc9c’, rf: 0.58 }, { name: ‘Magenta’, color: ‘#e84393’, rf: 0.48 }, { name: ‘Yellow’, color: ‘#f1c40f’, rf: 0.78 } ]; } else if (solvent === ‘ethanol’) { data = [ { name: ‘Cyan’, color: ‘#1abc9c’, rf: 0.52 }, { name: ‘Magenta’, color: ‘#e84393’, rf: 0.40 }, { name: ‘Yellow’, color: ‘#f1c40f’, rf: 0.68 } ]; } else { data = [ { name: ‘Cyan’, color: ‘#1abc9c’, rf: 0.16 }, { name: ‘Magenta’, color: ‘#e84393’, rf: 0.12 }, { name: ‘Yellow’, color: ‘#f1c40f’, rf: 0.20 } ]; } } return data; }function initSpots() { var data = getRfData(); spots = []; for (var i = 0; i PAPER_TOP + 12) { solventY -= riseSpeed * dt; if (solventY 1.4) { initSpots(); } return; }var solventTravel = BASELINE – solventY; if (solventTravel < 0) solventTravel = 0;for (var i = 0; i 3.5) { s.trail.push(s.y); if (s.trail.length > 35) s.trail.shift(); } }simTime += dt; updateResult(); }function updateResult() { var solventTravel = BASELINE – solventY; if (solventTravel < 1) solventTravel = 1; var parts = []; for (var i = 0; i < spots.length; i++) { var dist = BASELINE – spots[i].y; var rf = (dist / solventTravel).toFixed(2); parts.push(spots[i].name + ': Rf ' + rf); } resultBox.innerHTML = 'Time ' + simTime.toFixed(1) + ' s  |  ' + parts.join('  ·  '); }function draw() { ctx.clearRect(0, 0, W, H); ctx.fillStyle = '#f5f0e6'; ctx.fillRect(0, 0, W, H);// paper area ctx.fillStyle = '#faf6ee'; ctx.beginPath(); if (ctx.roundRect) ctx.roundRect(PAPER_L, PAPER_TOP, PAPER_R – PAPER_L, PAPER_BOT – PAPER_TOP, 6); else ctx.rect(PAPER_L, PAPER_TOP, PAPER_R – PAPER_L, PAPER_BOT – PAPER_TOP); ctx.fill();// texture ctx.strokeStyle = 'rgba(180,160,120,0.12)'; ctx.lineWidth = 0.6; for (var y = PAPER_TOP + 10; y < PAPER_BOT; y += 12) { ctx.beginPath(); ctx.moveTo(PAPER_L + 5, y); ctx.lineTo(PAPER_R – 5, y); ctx.stroke(); }// baseline ctx.strokeStyle = 'rgba(80,60,40,0.4)'; ctx.lineWidth = 1.2; ctx.setLineDash([4, 3]); ctx.beginPath(); ctx.moveTo(PAPER_L + 6, BASELINE); ctx.lineTo(PAPER_R – 6, BASELINE); ctx.stroke(); ctx.setLineDash([]); ctx.fillStyle = 'rgba(80,60,40,0.55)'; ctx.font = '10px sans-serif'; ctx.textAlign = 'left'; ctx.fillText('baseline', PAPER_L + 8, BASELINE – 5);// solvent reservoir ctx.fillStyle = 'rgba(90,160,240,0.28)'; ctx.fillRect(PAPER_L, SOLVENT_START – 2, PAPER_R – PAPER_L, PAPER_BOT – SOLVENT_START + 2);// solvent front + wet zone if (solventY < SOLVENT_START – 1) { var grad = ctx.createLinearGradient(0, solventY, 0, BASELINE + 5); grad.addColorStop(0, 'rgba(100,175,255,0.2)'); grad.addColorStop(1, 'rgba(100,175,255,0.03)'); ctx.fillStyle = grad; ctx.fillRect(PAPER_L + 2, solventY, PAPER_R – PAPER_L – 4, BASELINE – solventY + 6);ctx.strokeStyle = 'rgba(50,130,220,0.75)'; ctx.lineWidth = 2; ctx.beginPath(); ctx.moveTo(PAPER_L + 3, solventY); ctx.lineTo(PAPER_R – 3, solventY); ctx.stroke();ctx.fillStyle = 'rgba(50,130,220,0.85)'; ctx.font = '10px sans-serif'; ctx.textAlign = 'right'; ctx.fillText('solvent front', PAPER_R – 6, solventY – 5); }// spots var centerX = (PAPER_L + PAPER_R) / 2; var offsets = [-42, 0, 42];for (var i = 0; i < spots.length; i++) { var s = spots[i]; var x = centerX + offsets[i];// trail ctx.globalAlpha = 0.22; for (var t = 0; t < s.trail.length; t++) { ctx.fillStyle = s.color; ctx.beginPath(); ctx.arc(x, s.trail[t], 3.5 + (t / s.trail.length) * 3, 0, Math.PI * 2); ctx.fill(); } ctx.globalAlpha = 1;// main spot ctx.shadowColor = s.color; ctx.shadowBlur = 10; ctx.fillStyle = s.color; ctx.beginPath(); ctx.arc(x, s.y, 9, 0, Math.PI * 2); ctx.fill(); ctx.shadowBlur = 0;ctx.fillStyle = 'rgba(255,255,255,0.35)'; ctx.beginPath(); ctx.arc(x – 2.2, s.y – 2.2, 2.8, 0, Math.PI * 2); ctx.fill();// label ctx.fillStyle = '#334155'; ctx.font = '10px sans-serif'; ctx.textAlign = 'center'; ctx.fillText(s.name, x, s.y – 14); }// faded origin marks ctx.globalAlpha = 0.3; for (var i = 0; i < spots.length; i++) { ctx.fillStyle = spots[i].color; ctx.beginPath(); ctx.arc(centerX + offsets[i], BASELINE, 4.5, 0, Math.PI * 2); ctx.fill(); } ctx.globalAlpha = 1; }function loop(now) { if (!last) last = now; var dt = Math.min((now – last) / 1000, 0.05); last = now; update(dt); draw(); requestAnimationFrame(loop); }function softReset() { initSpots(); updateResult(); }resetBtn.addEventListener('click', function () { softReset(); running = true; }); pauseBtn.addEventListener('click', function () { running = false; }); playBtn.addEventListener('click', function () { running = true; last = performance.now(); }); solventSelect.addEventListener('change', softReset); mixtureSelect.addEventListener('change', softReset);initSpots(); updateResult(); requestAnimationFrame(loop);// Quiz var submitQuiz = document.getElementById('submitQuizBtn'); var quizResultDiv = document.getElementById('quizResult'); var questions = document.querySelectorAll('.question');submitQuiz.addEventListener('click', function () { var score = 0; var total = questions.length; for (var idx = 0; idx < questions.length; idx++) { var q = questions[idx]; var correctVal = q.getAttribute('data-correct'); var selected = q.querySelector('input[type=radio]:checked'); var labels = q.querySelectorAll('.options label'); var expDiv = q.querySelector('.explanation'); for (var li = 0; li < labels.length; li++) { labels[li].classList.remove('correct'); labels[li].classList.remove('wrong'); } if (selected) { if (selected.value === correctVal) { score++; selected.parentElement.classList.add('correct'); } else { selected.parentElement.classList.add('wrong'); } } for (var li = 0; li = 80) msg += ‘Excellent mastery of chromatography!’; else if (percent >= 60) msg += ‘Good! Review the Rf calculation and types.’; else msg += ‘Please review the principles and simulation again.’; quizResultDiv.innerHTML = msg; quizResultDiv.style.padding = ’12px’; quizResultDiv.style.background = ‘#e6f0f5′; quizResultDiv.style.borderRadius = ’20px’; submitQuiz.textContent = ‘Retake Quiz’; submitQuiz.onclick = function () { location.reload(); }; }); })();

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