This website is written by experts in Pharmaceutical Industry Manufacturing Research and Development and Regulatory Affairs Since 2008 and in 2011 Awarded as one of top 10 websites across world in Pharmaceutical Manufacturing Regulatory
USFDA WHO GMP Guidelines Pharmaceutical Regulations Information about Pharma Quality Managment System Validations Process Validation Cleaning Validation Equipment Qualification Equipment Process Validation Computer System Validation Sterile Product Manufacturing Guidelines Media fill guideline Aseptic Techniques in Sterile dosage form Clean Room Clean Room Classification Guidelines on Area Qualification HVAC AHU HEPA Filter HEPA Filter integrity testing and leak test

Sunday, December 13, 2009

Chromatographic Separation Of Purin Bases

How to Do Chromatographic Separation Of Purin Bases
Chromatography is an analytical method and technique of separation in which the ingr. Are separated on basis of their difference in migration in a system of two phases. One is fixed on a stationary phase and other is a mobile phase. The difference in migration rates of ingr. Are based on different adsorption partition on exchange molecular sieving effect chief subst. to be separated.
The stationary phase is solid absorbent or a lia. Feed on a solid carrier. Mobile phase travel through stationary phases it carries with it ingr. From sample at different rates. The technique is widely used for separation of amino acids, lipids, alkaloids etc. TLC of Purina bases viz. adenine and Uralic stationary phase: silica gel supported as glass plate.


Mobile phase: - n-butane, n-methanol, H2O, NH3 in ratio 60:20:2:1.
a) Purina bases show weak flu ore séance in U.V.light.
b) Spray of 0.2% cosign in ethanol sate. C mercuric chloride which forms a complex with bases is appearing as black spot in U.V.light.


Procedure:- Preparation of plate: Prepare 10% slurry of silica gel in H2O and spread it evenly on a glass plate and allow it to set – dry it at 1100c and wol.


Soln of chamber: - Preparation a solvent ys. And pour about 20 ml in a TLC chamber and put a pica of filter paper in it close in air tight container keep it from nearly 30 mins.

Preparation of sample: - Prepare 1% w/w soln of adenine and Uralic in hot water and sy of them.
Spotting: Spot about 0.05 ml of each sample and each std. soln on TLC and dry in air at r.t.note that spots don’t spread two much are sufficient apart.
Developing: - Pat the plate. In previously sold chamber and allow to develop till the solvent runs to a distance of 20 cms. Mark the solvent front, take out the plates and keep as flat sur. Dry in air.
Location: - Observe the plates as such under UV or spray the plate c reagent. Allow to stand for 15 min and observe under UV the spots and calculate RP value.


Observation:-
Dist. Traveled by A = 10 cms.
Dist. Traveled by B = 8 cms.
Dist. Traveled by solvent = 15 cms.


Rf = Dist.travelled by samples .
Dist.travelled by solvent front.


Therefore Rf of A = 10/15 = 0.66


Rf of B = 5 /15= 0.53


Result:-
Rf of A = 0.66
Rf of B = 0.53
========================================================================
This website is a guide for for pharmacy students , pharmaceutical professional working in pharmaceutical regulatory affairs , pharmaceutical manufacturing , quality assurance , pharmaceutical formulation development , research and development.
This week we are publishing a series of articles on biochemistry, over this website , for our pharma students world wide .
We recomend you to sebscribe to this website and receive our articles directly by email when ever we update this website.

This website is most favorite Amongst proffesionals in almost all pharmaceutical companies in USA , Pharmaceutical companies in UK , Pharmaceutical companies in Japan , Canada , UAE , Pharmaceutical companies in India 

Following are some of articles which;will be useful for you in further understanding of aspects of sterile dosage form manufacturing

Pharmaceutical Validation

Cleen Room Classification

Pharmaceutical Industry

Pharmaceutical Manufacturing

Clinical Trials

Media fill

Regulatory Affairs in Pharmaceutical Industry

No comments:

Pharma Guidelines Most Read

.

Readers Visit Number

This website is Authentic Source of Pharmaceutical Guidelines

This website is Authentic Source of Pharmaceutical Guidelines

Pharma Validations Quality Management System USFDA Guidelines WHO GMP Guidelines New Drug Update





This website consist of detail Information about Pharma Guidelines Pharma Regulatory Affairs USFDA Guidelines WHO Guidelines Pharmaceutical Quality Management System Pharma Validations Process Validation Cleaning Validation Equipment Qualification Equipment Process Validation Computer System Validation Sterile Product Manufacturing Guidelines Media fill guideline Aseptic Techniques in Sterile dosage form Clean Room Clean Room Classification Guidelines on Area Qualification HVAC AHU HEPA Filter HEPA Filter integrity testing and leak test