PKKK/300/UP/062 Determination of Volatile Compounds in Cosmetic Products using GCMS

GCMS

Ultrasonic bath

Glass conical flasks with screw cap

Amber volumetric flask (25 mL, 50 mL)

Nylon filter, 47 mm diameter, 0.45 µm pore size or equivalent

GC autosampler vial

Methanol

Menthol, Methyl Salicylate, Camphor, Thymol and Eugenol Reference Standard

Chromatographic Parameters

◼ Column: DB/BP-624, 6% Cyanopropylphenyl 94% Dimethylpolysiloxane, 30 m x 0.25 mm, 1.4 µm or equivalent

◼ Oven temperature:

Mode : EI

Injection volume : 0.5 ⎧L

Split ratio : 30:1

Column flow : 1.7 mL/min

Inlet temperature : 230°C

Interface temperature : 230°C

MS source temperature : 200°C

Full Scan Range : m/z 50 to m/z 200

Gas : Helium

SIM Mode settings : As below

Standard preparation:

Internal Standard (IS) Stock Preparation

Reference Standard Stock Preparation

Note: The weight of reference standard can be modified to minimize the usage of the reference standard.

Limit of Detection (LOD) Solution Preparation

Pipette 2.5 µL of Mixed Standard Solution and 200 µL of internal standard stock solution to 2 mL amber vial. Add 797.5 uL methanol. (LOD solution - Concentration of each compound 0.001mg/mL (1 ppm)

Test Method:

Standard Calibration Curve Preparation

Note: Alternative suitable dilutions to achieve desired concentrations can be used.

Sample Preparation (Liniment/Ointment/Cream/Solution)

Note: Appropriate dilution of sample should be made if the concentration of sample falls above the linear range of the calibration curve.

Cut sample into small pieces. Weigh 100 mg sample and add 15 mL methanol into a 50 mL conical flask, vortex for about 1 minute and sonicate with heat on a sonicator bath at 55 °C for 15 minutes. Cool the solution at room temperature and add 5 mL with IS Solution and make up to 25 mL with 5 mL methanol. Filter the sample solution using 0.45 µm Nylon filter into a 2 mL vial and cap. Prepare duplicates.

Note: Appropriate dilution of sample should be made if the concentration of sample falls above the linear range of the calibration curve.

Measurement

System Suitability Test (SST) Quantitation Test:

The Tailing Factor of peaks not more than 2.0

The Theoretical Plate not less than 2000

R2, Coefficient of Determination from calibration curve is not less than 0.995.

Results Interpretation

Identification of active compound(s) is based on the following specification(s):

The results are interpreted as following according to specification(s) fulfilled:

If peak area ratio of compound to IS in sample solution is greater than the peak area ratio of standard solution at LOQ level, the amount of compound needs to be confirmed using extrapolation of calibration curve and assay calculation as below.

Assay amount of menthol, camphor, methyl salicylate and thymol (%w/w). The amount of menthol, camphor, methy salicylate and thymol present in the sample will be reported as percent weight/weight (%w/w). Determine the amount of menthol, camphor, methyl salicylate and thymol in the portion of sample taken using the following formula: