🇲🇾 NPRA — Pusat Kawalan Kualiti · ISO/IEC 17025 Accredited

◈ Seksyen Pengujian Produk & Kosmetik · Unit Penyaringan

PKKK/300/UP/052 Screening of Clindamycin by HPLC

Arahan Kerja Rasmi (Level 300 SOP) · Kategori: HPLC / LC

1. Reagen & Piawai

Penyediaan fasa bergerak, stok & kalibrasi

➔

2. Pengekstrakan

Timbang, sonikasi, sentrifug & turas 0.45µm

➔

3. Persediaan Alat

Purging saluran, autotune & baseline

➔

4. Suntikan Batch

Blank, SST (n=6), Sampel & IQC

➔

5. Verifikasi & Rekod

Padanan RT, spektrum UV/MS & Laporan

⚡ Spesifikasi & Parameter Kromatografi / Analitikal (Official Parameters):

Instrumen UtamaHPLC Shimadzu Prominence-i LC-2030C

Turus / ColumnZorbax SB-C18 (150 × 4.6 mm, 5 µm)

Fasa Bergerak / PelarutBuffer Fosfat pH 3.0 : Acetonitrile (55:45 v/v)

Kadar Alir & Suhu1.0 mL/min @ 30 °C

Pengesan / Gelombang / SIM210 nm

Kriteria Kesesuaian Sistem (SST)SST %RSD ≤ 2.0% (n=6), Tailing T ≤ 2.0, Plates N ≥ 2000.

🎯 Sebatian Sasaran Pengujian:

Clindamycin

💡 Petua & Perhatian Penting Makmal (Bench Notes):

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.

Analyst Benchtop Checklist Progress

0 / 12 Langkah Selesai (0%)

NPRA PUSAT KAWALAN KUALITI	ARAHAN KERJA: SCREENING OF CLINDAMYCIN BY HPLC
	No. Dokumen: PKKK/300/UP/052	Terbitan / Semakan: Terbitan 3 Semakan 0
	Tarikh Kuatkuasa: 10 April 2026	Bahagian / Unit: SPPK · Unit Penyaringan

0.0 SEJARAH PINDAAN & SEMAKAN 1.0 OBJECTIVE 2.0 SCOPE 4.0 FLOW CHART 6.0 PROCEDURE 6.0 PROCEDURE 7.0 QUALITY RECORDS 8.0 ATTACHMENTS 📊 Jadual Data (2)

📋

0.0 SEJARAH PINDAAN & SEMAKAN

AMENDMENT :

This procedure is a new document in line with upgrading the institution’s certification to the ISO 9001: 2015 version and the renaming of the institution from National Pharmaceutical Control Bureau (NPCB) to Bahagian Regulatori Farmasi Negara (NPRA). Therefore, the original document is obsolete and this document is published as Version 1.

HISTORY OF REVISION :

Issue 2 Revision 0 :

Update on document number from PKK/200/KOS/022 to PKK/200/HMS/010

Update on effective date from 15 November 2016 to 10 February 2020

Update on responsibility.

Issue 3 Revision 0 :

Document change from testing procedure PKK/200/HMS/010 to work instruction PKK/300/HMS/065

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 :

Update document number from PKK/300/HMS/065 to PKKK/300/HMS/065

Update paragraph 5.0 Responsibility

Issue 5 Revision 0 :

i. Update document number from PKKK/300/HMS/065 to PKKK/300/KOS/022

ii. Update paragraph 5.0 Responsibility

Update paragraph 7. Quality Records

Issue 5 Revision 1:

i. Update document number from PKKK/300/KOS/022 to PKKK/300/UP/052

📋 Sejarah Pindaan & Semakan Dokumen (5 entri) ▸

ii. Pinda tarikh kuatkuasa 1 April 2024 kepada 10 April 2026.

iii. Kemaskini para 7.0 Quality Records :

KOS/X003 kepada UP/X009

KOS/X009 to UP/X013

KOS/002A kepada UP/002B

🎯

1.0 OBJECTIVE

◼ To screen the presence of clindamycin in cosmetic products using HPLC.

📐

2.0 SCOPE

◼ This test method is only to be used for screening of clindamycin in cosmetic products which is suspected to contain the substance.

3.0 Definition

3.1 HPLC : High Pressure Liquid Chromatography

3.2 PTFE : Polytetrafluoroethylene

3.3 Analyst: Pharmacist/Pharmacy assistant/Science Officer

🔀

4.0 FLOW CHART

▹ - None -

Responsibility

Head of Section

5.1.1 Validate analysis report of failed sample and approve on QUEST System

Head of Unit / Pharmacist in Charge

Distribute samples for testing to Pharmacist through QUEST system

Validate and approve analysis report on QUEST system

Review investigation report of failed samples

Pharmacist

Distribute samples for testing among analysts

Ensure tests are conducted according to laboratory protocols

Review analysis reports and update into the QUEST system

Investigate and report failed samples

Perform sample analysis (when necessary) and report

Pharmacy assistant / Analyst

Perform sample analysis and report

🔬

6.0 PROCEDURE

6.1 Equipment/ Materials

High Pressure Liquid Chromatograph with a variable visible wavelength detector

Whatman PTFE disposable syringe filter, 13 mm diameter, 0.45 µm pore size or equivalent

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

Ultrasonic bath

Langkah 1 Water bath

10ml volumetric flask

20ml volumetric flask

Langkah 2 Filter paper

Methanol (HPLC grade)

Sodium Dihydrogen Phosphate (NaH2PO4)

Water used must be Ultrapure water, 18 mega ohm

Orthophosphoric acid 85% (H3PO4)

0.01M Phosphate Buffer Solution adjusted with orthophosphoric acid to pH 2.5

Solvent: Mobile phase

Reference material: Clindamycin phosphate standard

Chromatographic Parameters

◼ Column : uBondapak C18 125A, 300 mm x 3.9 mm i.d., 10 µm

◼ System : Isocratic

◼ Mobile phase : 0.01M Phosphate buffer solution pH 2.5/methanol with 50:50 (v/v)

◼ Temperature : 40oC

◼ Flow rate : 0.8 mL/min

◼ Detection : UV, 205 nm

◼ Injection volume : 20 μl

Preparation of 0.01M Phosphate buffer solution ph 2.5

Langkah 3 Weigh ~10.54g NaH2PO4 and dilute with 500mL of water. Adjust pH to 2.5 with orthophosphoric acid

🔬

6.0 PROCEDURE

Reference Standard Preparation

Langkah 1 Weigh accurately 5 mg of the Clindamycin Phosphate Reference Standard and make up to volume 20 mL with solvent. Sonicate for 5 minutes. Filter the solution before injecting into the HPLC.

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

the usage of the reference standard.

Sample Preparation

Langkah 2 Weigh accurately 0.5 g of the sample into a 20 mL volumetric flask and fill the mark with solvent. Sonicate 30 minutes. Filter the solution before injecting into the HPLC.

Note: The weight of sample can be modified due to the availability and situation of the sample received.

Measurement

Langkah 3 Inject 20 μl Reference Standard solutions and sample preparation into the HPLC and check for the chromatogram.

Results

Compare the retention time obtained from the sample chromatogram with that of the Clindamycin Phosphate standards.

The identity test is positive if the retention time of the sample is identical to that obtained from the standard chromatogram.

📝

7.0 QUALITY RECORDS

Rumusan Laporan Pengujian Kromatografi : UP/X009

Keputusan Ujian Kosmetik : UP/X013

Laporan Siasatan Sampel Tidak Lulus Ujian Kromatografi : UP/002B

📎

8.0 ATTACHMENTS

Chromatogram of Reference Standard

Chromatogram of System Suitability test

Chromatogram of Reference Standard

Chromatogram of System Suitability Test

📊

Jadual Kromatografi & Rujukan Data

JADUAL 1 PKKK/300/UP/052

HISTORY OF REVISION	HISTORY OF REVISION	HISTORY OF REVISION	HISTORY OF REVISION	HISTORY OF REVISION	HISTORY OF REVISION
Issue	Revision	Written by	Reviewed by	Approved by	Effective date
1	0	Nor Izyani Hanafi	Ooi Suat Hong	Dr Noraida Mohamad 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	Gopinatha Ganesh	Tan Lu Yi	Ida Syazrina Ibrahim	10 April 2026

JADUAL 2 PKKK/300/UP/052

REFERENCE	REFERENCE
Document No.	Title
	USP Monograph 33