1.0 OBJECTIVE:To lay down the procedure for issuance, maintenance & dispose of HPLC / GC Column.
2.0 SCOPE:This SOP shall be applicable for use of new column and, maintenance and retrieval of old HPLC/GC columns.
3.0 RESPONSIBILITY:3.1 QC Executives/officer indent and retrieval of column
4.0 ACCOUNTABILITY:HOD QC/ Head QA/QC
5.1 Indent & usages of new HPLC column5.1.1 QC executive shall indent the required HPLC/GC column and send to purchase department for procurement after approval of Head QC/QA.
5.1.2 QC person shall received the new column & verified the physical verification of column against indent, bill .
5.1.3 QC person shall conditioning the column as per define procedure & check the performance of new column as follow step after conditioning of column:
220.127.116.11 Wash the Column with 100% with degas Solvent such as HPLC grade Methanol /Acetonitrile for 30 min.
18.104.22.168 Wash the Column 100% with degas HPLC /filtered & degas distilled water up to 30 min.
22.214.171.124 Wash the Column with 60:40 ratio of degas TKA/Milli Q water and solvent up to 30 min.
126.96.36.199 Now check the performance of column as per given method prescribe & directed by manufacturer provide with column OR check the performance of column with first injection of standard, should be meet the system suitability criteria as given in respective method of analysis of product.
188.8.131.52 Chiral Column to be condition as per manufactures guidelines.
184.108.40.206 Washing or conditioning of proteinus, silica based or chiral column should not used water, ratio of low polarity solvent & water can be use for conditioning of this type of column or as per method directed by manufacturer.
5.1.4 QC person shall assign I.D.No. of column if performance of column found satisfactory .
5.1.5 QC person shall take the entry of new column in issuance and destruction register & acknowledge receipt and send the duplicate copy to purchase dept if performance of column found satisfactory .
5.1.5 If performance of column does not meet with in-house/ manufacturer criteria then immediate inform to QC HOD, purchase department and supplier to replace the column.
5.2 Performance and re- generation of in-use column5.2.1 Analyst shall rewash the column with 100% degas water 1hr & then ratio of water & solvent (60:40 water: solvent) 1 to 2 hour if performance or system suitability of column fails during routine analysis .
5.2.2 QC person shall check the performance of column after washing with first injection of standard as per given method of analysis, should be meet the system suitability criteria as given in method of analysis of respective product.
5.2.3 Column to be regenerated if found poor performance after rewash with Acetonitrile/Methanol/THF/Butanol/IPA as per requirement & description of column and then wash with 60:40 Ratio of degas water: solvent 1-2 hr.
5.2.4 For Protein us column regenerated with 50:25:25 ratio of Water: IPA: Ethanol up to 30 min and then 80:20 ratio of water: acetonitrile up-to 20min and chiral column to be re-generate with Ethanol/Hexane or as directed by manufacturer/supplier
5.2.5 QC Head shall retrieve and dispose failure old column with detail entry in issuance and Destruction register and take in own custody to avoid reuse of column if it is not meeting the criteria of tailing factor, theoretical plates, resolution, fronting, splitting / damage etc.
5.2.6 After disposal of old column QC head issue the new column with given identification number of new column.
5.3 For GC column5.3.1 Follow the same procedure for indent & issue of column as mentioned in receipt of HPLC column.
5.3.2 Condition the issued new column up to 4-5 hour at 220°C oven temperature and calculate the theoretical plate and resolution of column should comply as given criteria in the method of analysis .
5.3.3 Immediate inform to QC manager if not meet the performance given by manufacture /supplier.
5.4 Cleaning and maintenance of column during analysis5.4.1 Analyst shall wash the in-use column with degas distilled water after every analysis up to 45 min and then 30 min wash with ratio of 60:40 water: solvent (solvent use which ever using in testing).
5.4.2 If using high concentration of buffer during analysis then wash the column by filtered 50°c hot water & then 100ml with degas solvent.
5.4.3 Analyst shall ensure that column should wash with mixture of methanol / acetonitrile or acetonitrile & methanol or as per the specification recommended by manufacturer if store for long time.
5.5 PrecautionsHow to protect LC column
- Degassed solvents, water and mobile phase should be used .
- Use clean filters.
- Flush several volumes of 100% methanol (or similar) through the HPLC system after water washing a prior to shutdown.
- Clean pump, Injector loop and syringe with hot water in twice a week.
- Filter sample to be injected to avoid deposition of contamination at column end.
- Column should store at room temp.
- Protect the column from knocks and bumps
5.6 Discard of HPLC / GC column5.6.1 Discard the column if it is not meeting the criteria of Tailing factor, theoretical plates, Resolution, Fronting, Splitting / damage etc.
5.6.2 QC person shall make the entry in the Column Issuance and Destruction record
5.6.3 Discarded columns shall kept separately and affixed the label as “Discarded column-Do not Use”.
6.0 ABBREVIATIONS:6.1 QA: Quality Assurance
6.2 QC: Quality Control
6.3 HPLC: High Pressure Liquid Chromatography
6.4 GC: Gas Chromatography
6.5 HOD: Head of Department
6.6 SOP: Standard Operating Procedure
6.7 IPA: Iso propyl Alcohol
Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
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