No.

930035-01


Test Procedure Page 1 of 4
Remarks This method corresponds to Q-C-2053.0 and to method CP-1185
Grnau



Author






- confidential information -














Content of - bisabolol in Hydagen B
by a gas chromatographic method
















No. 930035-01

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Test Procedure
1 Purpose and field of application

The method is used to determine -bisabolol in Hydagen. It is suitable for the range 0-
100%.



2 Principle

The sample is dissolved in pyridine, separated by means of capillary gas
chromatography, and detected and quantified with the FID. The quantification is in terms
of percentage area.



3 Reagents

Pyridine, p.a. (e.g. Merck art. no. 7463)



4 Apparatus

Gas chromatograph (e.g. Siemens Sichromat 1)
Gas sampler (e.g. Siemens AS 200)
FID (e.g.: Siemens
Injector split (e.g. Siemens)
Column DB 5, 30m 0.32mm ID, 0.1m film thickness (e.g. J&W)



5 Experimental


5.1 Sample preparation

Mix the sample thoroughly, then weigh approx. 20mg accurately to within 0.1mg in an
ampoule and dissolve in 1ml pyridine.

Dilute the sample solution 1:5 and inject into the gas chromatograph with the help of a
gas sampler.
Carry out a double determination of each sample.
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Test Procedure
5.2 GC parameters

Carrier gas: Hydrogen 5.0
Carrier prepressure: 1 bar (approx. 7ml/min)
Injector temperature: 340C
Detector temperature: 350C
Split: 190ml/min
Temperature program: Initial temperature: 80C
Start time: 0 min
Rate: 6C/min
End temperature: 340C
End time: 0 min
FID amplifier: Det. 3
Make up GAS: Synth. air 55ml/min
Fuel gas: Synth. air 50ml/min (preliminary pressure 2.2 bar)
Integrator: Shimadzu CR 3A/Cr 4A SP 4290 with "Winner" system



6 Evaluation

The - bisabolol content is calculated as a percentage area.

A * 100
% - bisabolol = ------------------------
B

A = Peak area of - bisabolol
B = Sum of all peak areas



7 Result

The result is rounded to one decimal place and mentioning this method.



8 Safety information

The method described involves handling hazardous substances. For this reason your
attention is drawn to the numerous regulations concerning the handling of hazardous
substances. Technical, organisational and personal protective measures must be
observed.
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Test Procedure


Figure 1: Typical chromatogram