PKKK/300/UP/050 Screening of ID Hydroquinone by HPLC 10 April 2026
- Tapis semua fasa bergerak (akueus dan organik) menggunakan penuras membran 0.45 µm dan degas dalam kukus ultrasonik selama sekurang-kurangnya 15–20 minit.
- Lakukan Auto Purge / Manual Purge setiap kali pelarut ditambah atau ditukar jenis bagi menyingkirkan buih udara.
- Pastikan garisan dasar (baseline) dan tekanan pam stabil (RSD < 2%) sebelum memulakan suntikan sampel kelompok.
- Bagi sampel matriks kapsul lembut (softgel), gunakan Chloroform kerana gelatin/minyak tidak larut dalam Methanol.
|
NPRA PUSAT KAWALAN KUALITI |
ARAHAN KERJA: SCREENING OF ID HYDROQUINONE BY HPLC 10 APRIL 2026 | |
| No. Dokumen: PKKK/300/UP/050 | Terbitan / Semakan: Terbitan 3 Semakan 0 | |
| Tarikh Kuatkuasa: 10 April 2026 | Bahagian / Unit: SPPK · Unit Penyaringan | |
0.0 SEJARAH PINDAAN & SEMAKAN
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
1 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
HISTORY OF REVISION
Issue
Revision
Written by
Reviewed by
Approved by
Effective date
1
0
Nor Izyani Hanafi
Ooi Suat Hong
Dr Noraida Mohd
Zainoor
15 November
2016
2
0
Nor Izyani Hanafi
Ooi Suat Hong
Wan Nurul Aina
Mior Abdullah
10 February
2020
3
0
Nor Izyani Hanafi
Ooi Suat Hong
Wan Nurul Aina
Mior Abdullah
15 July 2020
3
1
Nor Izyani Hanafi
Ooi Suat Hong
Wan Nurul Aina
Mior Abdullah
15 Disember
2020
4
0
Gopinatha
Ganesh
Nor Izyani Hanafi
Wan Nurul Aina
Mior Abdullah
1 November
2022
5
0
Leong Lip Wai
Gopinatha Ganesh
Nor Izyani Hanafi
1 April 2024
5
1
Leong Lip Wai
Gopinatha Ganesh
Nor Izyani Hanafi
1 Julai 2024
5
2
Nur Syazana
Ahmad Nordin
Leong Lip Wai
Nor Izyani Hanafi
2 Januari 2025
5
3
Leong Lip Wai
Nur Syazana
Ahmad Nordin
Nor Izyani Hanafi
23 Jun 2025
5
4
Leong Lip Wai
Nur Syazana
Ahmad Nordin
Nor Izyani Hanafi
13 Ogos 2025
6
0
Nurul Nadiah Noh
Gopinatha Ganesh
Ida Syazrina
Ibrahim
10 April 2026
REFERENCE
Document No.
Title
Standard Operating Procedure for HPLC Agilent
PKKK/300/KOS/013
Standard Operating Procedure for HPLC Shimadzu Series LC-
20AT
Journal of Food and Drug Analysis, Vol.12,No.1,2004
AMENDMENT:
This procedure is a new document in line with upgrading the institution’s certification to ISO
9001:2015 version and renaming of the institution from National Pharmaceutical Control
Bureau (NPCB) to National Pharmaceutical Regulatory Agency (NPRA). Therefore, the
original document is obsolete and this document is published as Issue 1.
HISTORY OF REVISION :
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
2 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
Issue 2 Revision 0 :
i.
Update on document number from PKK/200/KOS/019 to PKK/200/HMS/007
ii.
Update on effective date from 15 November 2016 to 10 February 2020
iii.
Update on responsibility.
Issue 3 Revision 0 :
i.
Document change from testing procedure PKK/200/HMS/007 to work instruction
PKK/300/HMS/062
Issue 3 Revision 1 :
i.
Update on name of agency from National Pharmaceutical Regulatory Agency
(NPRA) to Bahagian Regulatori Farmasi Negara (NPRA)
Issue 4 Revision 0 :
i.
Update title of document to Screening for Identification of Hydroquinone in Cosmetic
Products using HPLC technique
ii.
Update document number from PKK/300/HMS/062 to PKKK/300/HMS/062
iii.
Update paragraph 1.0 Objective
iv.
Update paragraph 2.0 Scope
v.
Update paragraph 3.0 Definition
vi.
Update paragraph 4.0 Flow Chart
vii.
Update paragraph 5.0 Responsibility
viii.
Update paragraph 6.0 Procedure
ix.
Update paragraph 7.0 Quality Records
x.
Update paragraph 8.0 Attachments
Issue 5 Revision 0 :
i.
Update document number from PKKK/300/HMS/062 to PKKK/300/KOS/019
ii.
Update paragraph 5.0 Responsibility
iii.
Update paragraph 7.0 Quality Records
Issue 5 Revision 1 :
i.
Update paragraph 6.4.2
ii.
Update paragraph 6.5.2 (a)
Issue 5 Revision 2:
i.
Amend document number from PKKK/300/HMS/053 to PKKK/300/KOS/013 and add
information in reference
ii.
Amend paragraph 5.0 Responsibility
iii.
Amend paragraph 6.0 Procedure
iv.
Omit flow chart in title Summary
Issue 5 Revision 3:
i.
Update paragraph 6.3.2 and 6.3.3
Issue 5 Revision 4:
i.
Update paragraph 6.5
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
3 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
Issue 5 Revision 4:
i.
Update effective date from 13 August 2025 to 10 April 2026
ii. Amend document number from PKKK/300/KOS/013 to PKKK/300/UP/050
1.0 OBJECTIVE
The test is performed to identify the presence of Hydroquinone in cosmetic cream
products by using HPLC.
2.0 SCOPE
The screening method is used for identification of Hydroquinone in cosmetic
cream products received from customers such as Surveillance & Complaint
Section, Pharmacy Enforcement and others.
IQC – Internal Quality Control
LOD – Limit of Detection
OOS – Out of Specification
Blank sample - Sample which has been tested before and does not contain
Hydroquinone
4.0 FLOW CHART
None
Responsibility
Head of Section
comply with specification.
Pharmacist in charge
procedure.
specification.
Pharmacist
Supervise testing conducted by the analysts.
Check test results and key in testing results in QUEST system.
does not comply with the specification.
Analysts
Perform sample testing according to the approved procedure
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
4 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
Prepare test reports
6.0 PROCEDURE
High Pressure Liquid Chromatograph with photo diode array detector
Ultrasonic bath
20 mL Volumetric flasks
50 mL capped Erlenmeyer flasks
Measuring cylinder
Nylon filter, 47 mm diameter, 0.45 µm pore size or equivalent
Methanol (HPLC grade)
Potassium Dihydrogen Phosphate (KH2PO4)
Water (Ultrapure water, 18 mega ohm)
Orthophosphoric acid 85% (H3PO4)
Diluent: 0.05 M Phosphate Buffer Solution pH 2.5
Reference material: Hydroquinone Reference Standard
Column
: C18 (Cosmosil 5 C18 AR-II), 250 mm x 4.6 mm i.d., 5 µm
System : Isocratic
Mobile phase
: 0.05M Phosphate buffer solution pH 2.5/ methanol with
99:1 (v/v)
Temperature
: 30 oC
Flow rate
: 0.9 mL/min
Detection
: UV, 280 nm
Injection volume
: 20 L
beaker and dissolve it with 300 mL of water. Transfer the solution into a 2 L
beaker. Make up to volume with water. Adjust the pH to 2.5 with
Orthophosphoric acid.
make up to volume 50 mL with diluent. (0.05 mg/mL of Hydroquinone).
Note : The reference standard stock solution is stable for 2 months when kept
in temperature between 2-8 °C
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
5 / 12
__________________________________________________________________________________________________________
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
Pipette 10 mL of Hydroquinone stock solution, make up to volume 20 mL with
diluent. (0.025 mg/mL of Hydroquinone). Filter the solution before injecting
into the HPLC.
Note : The reference standard working solution is stable for 2 months when
kept in temperature between 2-8 °C
Pipette 200 µL of Hydroquinone stock solution to 0.5 g of blank sample and
make up to volume 20 mL with diluent. (0.0005 mg/mL or 0.002 %w/w).
Note: The weight of reference standard can be modified to minimize the usage of the
reference standard.
flask. Measure 20 mL of diluent using measuring cylinder and pour into
Erlenmeyer flask. Sonicate for 30 minutes. Filter the solution before injecting
into the HPLC.
No. of
Injection
Injection ID
1
Blank / diluent
20
2
LOD solution
20
3
Reference standard
working solution
(Check for SST)
20
4
LOD solution
20
9
Sample solution
20
10
LOD solution
20
11
Sample solution
20
15
Sample solution
20
16
LOD solution
20
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
6 / 12
__________________________________________________________________________________________________________
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
ii) Theoretical plates should be NLT 1500
LOD solution (6.4.1) is injected in every 5 samples and at the end of analysis
according to its respective dosage form. Percentage difference of retention
time for LOD Solution is calculated by taking the retention time of before and
end of LOD Solution injected in every batch of 5 samples/end of analysis. The
percentage difference of retention time for LOD Solution should not be more
than 3 %.
sample chromatogram with that of Hydroquinone working standards
chromatogram.
solution.
standard.
7.0 QUALITY RECORD
Borang Tatacara Pengujian menggunakan kaedah HPLC : KOS/X004
ii)
Laporan Siasatan Sampel Tidak Lulus Ujian Kromatografi : KOS/002A
iii)
Borang Keputusan Ujian Sampel Kosmetik
: KOS/X009
% Difference = Difference in retention time of 2 LOD Solution x 100
Average of retention time of the respective 2 LOD Solution
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
7 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
8.0 ATTACHMENTS
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
8 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
ii)
Example of chromatogram for diluent, LOD solution, Hydroquinone working
standard & sample solution
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
9 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
iii)
Example of peak purity & spectrum for Hydroquinone working standard
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
10 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
11 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE
iv)
Example of peak purity & spectrum for Hydroquinone positive sample
NPRA
Document No:
PKKK/300/UP/050
Screening for Identification of
Hydroquinone in Cosmetic Products
using HPLC Technique
12 / 12
__________________________________________________________________________________________________________
CHECK ‘MASTER LIST’
ENSURE THAT THIS DOCUMENT IS THE CORRECT VERSION PRIOR TO USE