Regression analysis

Aim: - To plot the calibration curve of sulphanilamide and determine the slope, intercept and correlation coefficient. Procedure:- Dissolved 100 mg of given sample of sulphanilamide in sufficient water to produce 100 ml. Diluted 10 ml of this solution to 100 ml with water. From this pipette out 5 ml, 7.5 ml, 10 ml, 12.5 ml and 15 ml separately and transfered in 100 ml flask. Made the volume with water. The above solution had given 5, 7.5, 10, 12.5, 15 /ml respectively. Recorded the absorbance of all prepared solution at 254 nm in a 1 cm path length using water as reference solution. Ploted the calibration curve, absorbance Vs concentration. Find out the slope, intercept and correlation coefficient by using following formula.

Y = mx +b
Where y= Response (absorbance) x= concentration ,m= slope ,b= intercept

1) 2)

Result:1) Slope 2) Intercept, b 3) Correlation coefficient, r Observation Table Sr.no Concentration (/ml) x Absorbance y x2 y2 xy

1. 2. 3. 4. 5. 6.

n=

x=

y=

x2=

y2=

xy=

4) Equation y

EXPERIMENT NO: 07
Study of Effect of substituents on the absorbance spectra of Benzene chromophore

Aim: To study the effect of substituents like amino (-NH2) carboxyl(-COOH) separately on benzene ring and together (p-amino benzoic acid) on the absorption spectrum of benzene. Principle: Absorption of light in UV and visible region of spectrum is due to the presence of chromophore in the absorbing molecule. Chromophoric group include multiple bonds (-ene, -yne, carboxyl group) etc in which electrons are loosely held ( electrons). Changes in absorption spectra of the molecule can be produced by fully saturated groups called auxochromes. They do not have characteristic absorption on their own but can modify the absorption of chromophore. Its presence brings about a shift of absorption band towards the longer wavelength of spectrum (bathochromic shift) eg; amino, sulphur, carboxylic group. Procedure: Following solutions are prepared in water: 1. 0.005% w/v of aniline (50 mg in 100 ml, 1 ml of this solution to 10 ml) 2. 0.001% w/v of benzoic acid (100 mg in 100 ml, 1 ml of this to 100 ml) 3. 0.005% w/v of p-amino benzoic acid (50 mg in 100 ml, 1 ml of this to 10 ml) Record the absorption spectra over the range of 210-320 nm using 1 cm cell and water as reference solution. Result:

Compound Benzoic acid Aniline p-amino benzoic acid Cinnamic acid

max

Inference:

EXPERMENT NO: 08
Estimation of Benzene in Isopropyl Alcohol by second derivative spectroscopic method
Aim:- To determine the amount of benzene present in given sample of isopropyl alcohol by second derivative spectroscopic method.

Principle:- Derivate spectroscopy involves transformation of absorption spectra, into first ,second or higher order derivative spectra. A first derivative spectrum is a plot of gradient of the absorption curve ( rate of change of absorbance with wavelength dA/d) against wavelength . A second derivative spectrum is a plot of curvature of absorption spectra (d2A/d2) against wavelength. If absorption follows Beer-Lambert relationship ,the second derivative at any wavelength is related to concentration by following expression . (d2A/d2) = d2A (1%, 1 cm) x c d/d2 where, A= Absorbance at wavelength . A(1%,1cm) = Specific absorbance at wavelength . c = Concentration of absorbing solute expressed as percentage w/v. d = Thickness of absorbing layer in concentration. Procedure:Solution 1=> Sample being examined (isopropyl alcohol) Solution 2=> Preparation of 0.0002% v/v benzene in sample. 2 ml of benzene is diluted to 100 ml with given sample of isopropyl alcohol. 1 ml of this solution diluted to 100 ml with given sample (IPA), 1 ml of the resultant is further diluted to 100 ml with given isopropyl alcohol. Record the absorbance of both solution 1&2 over a range of 230 to 300 nm in 2 nm intervals by using 1 cm cell. Plot curvature of absorbance (d2A/d2) against wavelength .Measure the amplitude in spectrum obtained with solution 1 and 2.

Result:-

Observation:

230 232 234 236 238 240 242 244

Abs

Abs

Abs/ nm

2 Abs

2 Abs/ nm2

2 Abs/ nm2 X 100

246 248 250 252 254 256 258 260 262 264 268 270 272 274 276 278 280 282 284 286 288 290 292 294 296 298 300 Abs Abs Abs/ nm 2 Abs 2 Abs/ nm2 2 Abs/ nm2 X 100