Franklin Ifeanyichukwu Uba

CHEM 7750 - Separation science

 LC detectors

• Refractive index Detector (RID)

• Fluorescence detector

• Evaporative light
scattering detector (ELSD)

• UV-Detector
• Electrolytic conductivity detector • Electrochemical detectors
http://hplc.chem.shu.edu/HPLC/index.html
 ELSD principle
• Elution
• Droplet formation
• Solvent Evaporation
• Light scattering
(Rayleigh)
• Detection at 45O
• Mass detector
R.P.W.Scott, Chrom-Ed Book series, Book 5, 2003.
 Rayleigh scattering
• Elastic collision

ɵ
High energy Particle
photon

ɵ = Scattering angle http://upload.wikimedia.org/wikipedia/en/6/69/Rayleigh_sunlight_scattering.png

• KE conservation
• Particles smaller than wavelength of light
ELSD Instrumentation

http://www.sielc.com/images/ELSD_Schematic.gif
http://www.chem.agilent.com/
 Cholesterol and COPs

http://www.scielo.cl/fbpe/img/bres/v36n3-4/fig01.gif
 Previous methods
• Thin Layer Chromatography
• Qualitative
• Loading capacity restrictions
• Prone to contamination
• Gas Chromatography
• High temperatures (inlet/oven)
• Volatile derivatives (quantitative efficiency)
• High Pressure Liquid Chromatography
• No derivatives
• Faster
• Better accuracy than GC and TLC
 Elution conditions
• Normal phase column – 250 mm x 46 mm i.d.
• 16 % Alumina / 84% Silica ( 5μm )
• Guard Column – 20 mm x 2 mm

• Gradient elution – 1.0 mL/min

• Dichloromethane (A) / Acetonitrile : Water (99.05:0.95) (B)
• 94:6 / A:B for 0 – 6 min
• 79:21/ A:B for 27 min
• Linear Gradient to 94:6 for 5 min

• ELSD detector : 75 OC; N2 1.5 L/min

ELSD optimization
Results
 Linearity
Area = k (conc.)m

m = coefficient of curvature in
conc./peak area curves

ln(area) = ln K + m ln(conc.)

Normalized peak areas versus

concentration (0.1 – 1.0 g)
 Conclusions

• Fast analytical technique

• MDL of 75 ng/injection
• Stable and reproducible detector
• No baseline shift due to solvent changes
 Acknowledgement
• Dr Gilman

• CHEM 7750 Classmates

Hashim Treyvon Danielle

Jon Akanksha Nyote

Suzana Paul Sung

Kumuditha Sudha