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GB/T 35954-2018 English PDF (GBT35954-2018)
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GB/T 35954-2018: Determination of 10 whitening and freckle-removing components in cosmetics -- High performance liquid chromatography
GB/T 35954-2018
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 71.100.70
Y 42
Determination of 10 whitening and freckle-removing
components in cosmetics - High performance liquid
chromatography
ISSUED ON. FEBRUARY 06, 2018
IMPLEMENTED ON. SEPTEMBER 01, 2018
Issued by. General Administration of Quality Supervision, Inspection and
Quarantine;
Standardization Administration of the People's Republic of
China.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative references ... 4
3 Method summary ... 4
4 Reagents and materials ... 5
5 Instruments and equipment... 6
6 Analytical procedures ... 6
7 Calculation of result ... 8
8 Recovery and precision ... 9
9 Allowable difference ... 9
Annex A (informative) Detection limit and limit of quantitation of 10 whitening
and freckle-removing components ... 10
Annex B (informative) Chinese name, INCI name, CAS accession number,
molecular formula, structural formula and relative molecular mass of the
reference substance ... 11
Annex C (informative) Liquid chromatogram of standard solution of 10
whitening and freckle-removing components ... 14
Determination of 10 whitening and freckle-removing
components in cosmetics - High performance liquid
chromatography
1 Scope
This Standard specifies reagents and materials, instruments and equipment,
analytical procedures, calculation of results, recovery and precision, allowable
difference, etc. for high performance liquid chromatography of 10 whitening and
freckle-removing components (magnesium ascorbyl phosphate, ascorbyl
glucoside, β-arbutin, kojic acid, nicotinamide, 3-O-ethylascorbic acid,
potassium methoxysalicylate, raspberry glucoside, dipotassium glycyrrhizinate,
4-butyl resorcinol) in cosmetics.
This Standard applies to the determination of whitening and freckle-removing
components in water-based, lotion, cream, gel, wax-based, powder-based
cosmetics.
For the detection limits and quantification limits of this Standard method, see
Table A.1 in Annex A.
2 Normative references
The following referenced documents are indispensable for the application of
this document. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any
amendments) applies.
GB/T 6682, Water for analytical laboratory use - Specification and test
methods
3 Method summary
The sample takes potassium dihydrogen phosphate solution as extraction
solvent. Perform direct ultrasonic extraction of water-based samples and oil-in-
water samples. Water-in-oil samples, wax-based, and powder-based samples
are dissolved and dispersed in dichloromethane, and then extracted with
ultrasonic solvent. Perform centrifugation, filtration through a 0.22μm water filter.
Inject solution into a liquid chromatograph equipped with a diode array detector
20.0mg/L, 50.0mg/L, 100.0mg/L.
5 Instruments and equipment
5.1 High performance liquid chromatograph equipped with diode array detector
(DAD).
5.2 Centrifuge. >9500r/min.
5.3 Analytical balance. resolutions are 0.0001g and 0.001g.
5.4 PH meter.
5.5 Vortex mixer.
5.6 Ultrasonic cleaner.
5.7 Water microporous membrane. 0.45μm.
6 Analytical procedures
6.1 Sample processing
6.1.1 Preparation of water-based, oil-in-water samples
Weigh 0.5g of (to the nearest of 0.001g) sample in a 50mL colorimetric tube.
Add 30mL of potassium dihydrogen phosphate solution (4.4). Conduct vortex
oscillation until the sample is completely dispersed. Perform ultrasonic
extraction for 20min. Transfer to a 50mL volumetric flask. Use potassium
dihydrogen phosphate solution (4.4) to set volume to the scale. After mixing
evenly, take an appropriate amount of solution into the centrifuge tube.
Centrifuge at 9500r/min for 5min. Take the upper aqueous phase solution after
centrifugation. Filter through a 0.45μm water filter, the filtrate is to be
determined.
6.1.2 Preparation of water-in-oil, wax-based, powder-like samples
Weigh approximately 0.5g of (to the nearest of 0.001g) sample in a 50mL
stoppered centrifuge tube. Add 2.5mL of dichloromethane (4.2). Vortex until the
sample is completely dispersed (if a small amount of sample is not dispersed,
it can be sonicated to complete dispersion). Add 20mL of potassium dihydrogen
phosphate solution (4.4). Perform ultrasonic extraction for 20min, vortexing for
1min, centrifugation at 9500r/min for 5min. Remove the supernatant into a
50mL volumetric flask. Add 20mL of potassium dihydrogen phosphate solution
(4.4). Vortex for 1min, centrifuge at 9500r/min for 5min, and combine the
supernatant into a 50mL volumetric flask. Use potassium dihydrogen
Take the mixed standard working solution (4.8) and determine according to the
chromatographic conditions (6.2.1). Taking the peak area (Y) as the ordinate
and the standard solution mass concentration (X, mg/L) as the abscissa, draw
a standard working curve. For the liquid chromatogram of the standard solution
of whitening and freckle-removing components, see Figure C.1 in Annex C.
6.2.3 Specimen determination
The sample solution to be tested (6.1) is sequentially injected into the liquid
chromatograph. Determine according to chromatographic conditions (6.2.1).
Record the retention time and peak area of the chromatographic peak.
Calculate the content of the target compound in the sample solution according
to the standard curve. The response of the measured object in the sample
solution shall be within the linear range of the standard curve. Specimen
exceeding the linear range may be diluted with potassium dihydrogen
phosphate solution (4.4) and then determined.
NOTE. In order to ensure the accuracy of specimen quantification, try to complete the test process within
12h.
6.2.4 Qualitative confirmation
Perform qualitative determination of sample by liquid chromatograph. When the
sample is being determined, if the retention time of the peak of the tested
whitening and freckle-removing component is consistent with the standard, and
the chromatogram of the sample after subtracting the background, the UV
absorption of the substance is consistent with the UV absorption of the standard,
it can be preliminarily determined that the sample contains the measured
whitening and freckle-removing component. If necessary, other methods are
needed to confirm.
6.3 Blank test
Except for not weighing samples, they are carried out according to the above
measurement conditions and procedures.
6.4 Parallel test
According to the above steps, perform parallel test determination of the same
sample.
7 Calculation of result
The content of whitening and freckle-removing component in the sample is
calculated by the following formula.
Get QUOTATION in 1-minute: Click GB/T 35954-2018
Historical versions: GB/T 35954-2018
Preview True-PDF (Reload/Scroll if blank)
GB/T 35954-2018: Determination of 10 whitening and freckle-removing components in cosmetics -- High performance liquid chromatography
GB/T 35954-2018
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 71.100.70
Y 42
Determination of 10 whitening and freckle-removing
components in cosmetics - High performance liquid
chromatography
ISSUED ON. FEBRUARY 06, 2018
IMPLEMENTED ON. SEPTEMBER 01, 2018
Issued by. General Administration of Quality Supervision, Inspection and
Quarantine;
Standardization Administration of the People's Republic of
China.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative references ... 4
3 Method summary ... 4
4 Reagents and materials ... 5
5 Instruments and equipment... 6
6 Analytical procedures ... 6
7 Calculation of result ... 8
8 Recovery and precision ... 9
9 Allowable difference ... 9
Annex A (informative) Detection limit and limit of quantitation of 10 whitening
and freckle-removing components ... 10
Annex B (informative) Chinese name, INCI name, CAS accession number,
molecular formula, structural formula and relative molecular mass of the
reference substance ... 11
Annex C (informative) Liquid chromatogram of standard solution of 10
whitening and freckle-removing components ... 14
Determination of 10 whitening and freckle-removing
components in cosmetics - High performance liquid
chromatography
1 Scope
This Standard specifies reagents and materials, instruments and equipment,
analytical procedures, calculation of results, recovery and precision, allowable
difference, etc. for high performance liquid chromatography of 10 whitening and
freckle-removing components (magnesium ascorbyl phosphate, ascorbyl
glucoside, β-arbutin, kojic acid, nicotinamide, 3-O-ethylascorbic acid,
potassium methoxysalicylate, raspberry glucoside, dipotassium glycyrrhizinate,
4-butyl resorcinol) in cosmetics.
This Standard applies to the determination of whitening and freckle-removing
components in water-based, lotion, cream, gel, wax-based, powder-based
cosmetics.
For the detection limits and quantification limits of this Standard method, see
Table A.1 in Annex A.
2 Normative references
The following referenced documents are indispensable for the application of
this document. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any
amendments) applies.
GB/T 6682, Water for analytical laboratory use - Specification and test
methods
3 Method summary
The sample takes potassium dihydrogen phosphate solution as extraction
solvent. Perform direct ultrasonic extraction of water-based samples and oil-in-
water samples. Water-in-oil samples, wax-based, and powder-based samples
are dissolved and dispersed in dichloromethane, and then extracted with
ultrasonic solvent. Perform centrifugation, filtration through a 0.22μm water filter.
Inject solution into a liquid chromatograph equipped with a diode array detector
20.0mg/L, 50.0mg/L, 100.0mg/L.
5 Instruments and equipment
5.1 High performance liquid chromatograph equipped with diode array detector
(DAD).
5.2 Centrifuge. >9500r/min.
5.3 Analytical balance. resolutions are 0.0001g and 0.001g.
5.4 PH meter.
5.5 Vortex mixer.
5.6 Ultrasonic cleaner.
5.7 Water microporous membrane. 0.45μm.
6 Analytical procedures
6.1 Sample processing
6.1.1 Preparation of water-based, oil-in-water samples
Weigh 0.5g of (to the nearest of 0.001g) sample in a 50mL colorimetric tube.
Add 30mL of potassium dihydrogen phosphate solution (4.4). Conduct vortex
oscillation until the sample is completely dispersed. Perform ultrasonic
extraction for 20min. Transfer to a 50mL volumetric flask. Use potassium
dihydrogen phosphate solution (4.4) to set volume to the scale. After mixing
evenly, take an appropriate amount of solution into the centrifuge tube.
Centrifuge at 9500r/min for 5min. Take the upper aqueous phase solution after
centrifugation. Filter through a 0.45μm water filter, the filtrate is to be
determined.
6.1.2 Preparation of water-in-oil, wax-based, powder-like samples
Weigh approximately 0.5g of (to the nearest of 0.001g) sample in a 50mL
stoppered centrifuge tube. Add 2.5mL of dichloromethane (4.2). Vortex until the
sample is completely dispersed (if a small amount of sample is not dispersed,
it can be sonicated to complete dispersion). Add 20mL of potassium dihydrogen
phosphate solution (4.4). Perform ultrasonic extraction for 20min, vortexing for
1min, centrifugation at 9500r/min for 5min. Remove the supernatant into a
50mL volumetric flask. Add 20mL of potassium dihydrogen phosphate solution
(4.4). Vortex for 1min, centrifuge at 9500r/min for 5min, and combine the
supernatant into a 50mL volumetric flask. Use potassium dihydrogen
Take the mixed standard working solution (4.8) and determine according to the
chromatographic conditions (6.2.1). Taking the peak area (Y) as the ordinate
and the standard solution mass concentration (X, mg/L) as the abscissa, draw
a standard working curve. For the liquid chromatogram of the standard solution
of whitening and freckle-removing components, see Figure C.1 in Annex C.
6.2.3 Specimen determination
The sample solution to be tested (6.1) is sequentially injected into the liquid
chromatograph. Determine according to chromatographic conditions (6.2.1).
Record the retention time and peak area of the chromatographic peak.
Calculate the content of the target compound in the sample solution according
to the standard curve. The response of the measured object in the sample
solution shall be within the linear range of the standard curve. Specimen
exceeding the linear range may be diluted with potassium dihydrogen
phosphate solution (4.4) and then determined.
NOTE. In order to ensure the accuracy of specimen quantification, try to complete the test process within
12h.
6.2.4 Qualitative confirmation
Perform qualitative determination of sample by liquid chromatograph. When the
sample is being determined, if the retention time of the peak of the tested
whitening and freckle-removing component is consistent with the standard, and
the chromatogram of the sample after subtracting the background, the UV
absorption of the substance is consistent with the UV absorption of the standard,
it can be preliminarily determined that the sample contains the measured
whitening and freckle-removing component. If necessary, other methods are
needed to confirm.
6.3 Blank test
Except for not weighing samples, they are carried out according to the above
measurement conditions and procedures.
6.4 Parallel test
According to the above steps, perform parallel test determination of the same
sample.
7 Calculation of result
The content of whitening and freckle-removing component in the sample is
calculated by the following formula.
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