Analysis Of USP Method <467> Residual Solvents On The Agilent 8890 GC .
Application NoteResidual SolventAnalysis of USP Method 467 Residual Solvents on the Agilent 8890GC SystemAuthorLukas Wieder, Jie Pan, andRebecca VeenemanAgilent Technologies, Inc.2850 Centerville RoadWilmington DE 19808AbstractThis Application Note highlights the use of the Agilent 8890 GC and the Agilent J&WDB-Select 624 UI for 467 and Agilent J&W HP-INNOWax columns in the detectionand confirmation of 467 residual solvents. The system meets all specificationsrequired in USP Method 467 , and demonstrates excellent repeatability acrossseveral injections.
IntroductionClass 1 and class 2 residual solventsmust be monitored and regulated, andthe method for the analysis of thesesolvents involves three procedures: HSS transfer lineProcedure A: Initial identificationand limit test using a G43 phasecolumn (in this case, Agilent J&WDB-Select 624 UI for 467) Procedure B: If above limit inprocedure A, perform a confirmationof peak identity and secondary limittest using a G16 phase column (inthis case, Agilent J&W HP-INNOWax) Procedure C: If above limit inprocedures A and B, perform aquantification using whichevercolumn provided fewer coelutionsThis Application Note analyzed residualsolvents listed in USP Method 467 withthe Agilent 8890 GC. The J&W DB-Select624 UI for 467 and J&W HP-INNOWaxcolumns were used in this analysis, andconfigured with dual flame ionizationdetectors (FIDs). Therefore, proceduresA and B could be run simultaneously withone injection.ExperimentalEquipmentAn 8890 GC was configured with asplit/splitless inlet (SSL) and dual FIDs,and sampling was performed usingan Agilent 7697A headspace sampler.An inert tee was used to split the flowequally between the two columns,with both columns leading directly toFIDs. Figure 1 shows the completeconfiguration.Chemicals and reagentsDimethyl sulfoxide (99.9 %) and water(HPLC grade) were purchased fromSigma-Aldrich.2FIDCH 1S/SInletFIDCH 2Column 1J&W DB-Select 624 UIfor 467Deactivatedfused silicaInert tee7697Aheadspace samplerColumn 2J&W HP-INNOWAX8890 GCFigure 1. Experimental setup using a dual-column, dual-FID configuration for the analysis of USP 467 residual solvents.ConsumablesTable 1. Consumables and part numbers.ConsumableDescriptionVials10-mL Clear crimp top headspace vials (p/n 5190–2285)SeptaAdvanced Green nonstick inlet septa (p/n 5183–4759-100)SplitterInert tee for capillary flow technology (p/n G3184-60065)FerrulesShort graphite for 0.1 to 0.32 mm columns, 10/pk (p/n 5080-8853)UltiMetal Plus flexible metal, for 0.32 mm fused silica tubing, 10/pk (p/n G3188-27502)Inlet liner2 mm, Splitless, straight, deactivated (p/n 5181–8818)Headspace transferline/pre-CFT columnDeactivated Fused Silica, 30 m 0.25 mm id 0.35 mm od (p/n 160-2255-30)Column 1J&W DB-Select 624 UI for 467, 30 m 0.32 mm, 1.8 µm (p/n 123-0334UI)Column 2J&W HP-INNOWax, 30 m 0.32 mm, 0.25 µm (p/n 19091N-113I)
Sample preparationSample preparation for the residualsolvent samples was performedaccording to the USP 467 protocol.Experimental parametersTable 2. System parameters for the analysis of residual solvents.GC system parameter8890 GCThree stock solutions of residualsolvents in DMSO were used:Carrier gasHelium constant flow mode, 2 mL/min on column 1Inlet typeSplit/splitless Residual Solvent Revised Method 467 Class 1 (p/n 5190-0490)Inlet temperature140 CModeSplit mode, split ratio 5:1Residual Solvent Revised Method 467 Class 2A (p/n 5190-0492)Oven40 C (hold 5 min) to 180 C at 18 C/min (hold 3 min)Column 1 flow2 mL/min in constant flow mode, column 2 flow controlled by column 1Residual Solvent Revised Method 467 Class 2B (p/n 5190-0491)FID (both channels)250 CAir400 mL/minH230 mL/min The sample preparation proceduresfor each of the three classes are listedbelow:Makeup (N2)25 mL/minHeadspace parameter7697A headspace samplerSample loop1 mLOven temperature85 CLoop temperature85 CTransfer line temperature100 CVial equilibration time40 minutesInjection duration0.5 minutesVial size10 mLVial shakingOn, level 2 (25 shakes/min)Vial fill modeDefault: flow to pressureVial fill pressure15 psiLoop ramp rate20 psi/minClass 2A solventsFinal loop pressure0 psi1.Loop equilibration time0.05 minutesClass 1 solvents1.One milliliter of stock solution vialplus 9 mL of DMSO diluted to 100mL with water2.One milliliter from step 1 diluted to100 mL with water3.Ten milliliters from step 2 diluted to100 mL with water4.2.One milliliter from step 3 with 5 mLof water into headspace vialOne milliliter of stock solution vial,diluted to 100 mL with waterSoftwareAgilent OpenLab CDS – Version 2.2One milliliter from step 1 with 5 mLof water into headspace vialClass 2B solvents1.One milliliter of stock solution vial,diluted to 100 mL with water2.One milliliter from step 1 with 5 mLof water into headspace vials3
Results and discussionFigures 2 to 7 illustrate the analysis ofclasses 1, 2A, and 2B residual solventmixes on the J&W DB-Select 624 UI for467 and J&W HP-INNOWax GC columns.Analysis of class 1 solvents meets thesignal-to-noise ratio (S/N) and resolutionrequirements on both the J&W DB-Select624 UI for 467 and J&W HP-INNOWaxcolumns.In addition to showing clearchromatography on both columns foreach class of solvents and consistentresults across many runs, there arerequirements described in USP 467 the analysis must meet.The area and retention time repeatabilitymeasurements (RSD%) were evaluatedon a set of 10 headspace vials. Tables 3to 5 list the RSD% obtained on theJ&W DB-Select 624 UI for 467 andJ&W HP‑INNOWax columns for class 1,2A, and 2B residual solvent mixes.The resulting RSD% values were lessthan 5.0 %, indicating high repeatabilityand stability of the column, the 7697Aheadspace sampler, and the 8890GC/FID system.Response (pA)Class 1 4.03.93.83.73.63.53.43.33.23.13.0Compound1. 1,1-Dichloroethene2. 1,1,1-Trichloroethane3. Carbon tetrachloride4. Benzene5. 1,2-Dichloroethane45213012345678910Retention time (min)11121314151617Figure 2. Chromatogram of the USP residual solvents class 1 standard solution resolved on a J&W DB-Select 624 UI for 467 GC column.4RT4.0187.5907.7818.0268.094
Response hloroethene1,1,1-TrichloroethaneCarbon 2.9523.7346.4582,31012345678910Retention time (min)11121314151617Figure 3. Chromatogram of the USP residual solvents class 1 standard solution resolved on a J&W HP-INNOWax column.Table 3. Repeatability (n 10) for class 1 residual solvents obtained on the J&W DB-Select 624 UI for 467 and J&W HP-INNOWaxcolumns.Area RSD (%) on J&WDB‑Select 624 UI for 467RT RSD (%) on J&WDB‑Select 624 UI for 467Area RSD (%) on J&WHP‑INNOWaxRT RSD (%) on oethane3.70.314.20.0921.43.610.057Carbon tetrachloride2.90.060Coelutes with1,1,1-trichloroethaneCoelutes ,2-Dichloroethane3.20.0593.20.018Compound5
Class 2A solvents 3.23.02.82.6Response leMethylene 21.0120.8110.60.215540.4862 3190012345678910Retention time (min)11121314151617Figure 4. Chromatogram of the USP residual solvents class 2A standard solution resolved on a J&W DB-Select 624 UI for 467 GC column. 1033.0Compound1. Methanol2. Acetonitrile3. Methylene chloride4. trans-1,2-Dichloroethene5. cis-1,2-Dichloroethene6. Tetrahydrofuran7. Cyclohexane8. Methylcyclohexane9. 1,4-Dioxane10. Toluene11. Chlorobenzene12. Ethylbenzene13. m-Xylene14. p-Xylene15. o-Xylene142.82.62.42.2Response ,6290012345678910Retention time (min)111213141516Figure 5. Chromatogram of the USP residual solvents class 2A standard solution resolved on a J&W HP-INNOWax 076.3696.8598.6797.3687.4987.6098.193
Table 4. Repeatability (n 10) for class 2A residual solvents obtained on the J&W DB-Select 624 UI for 467 andJ&W HP-INNOWax columns.Area RSD (%) on J&WDB‑Select 624 UI for 467RT RSD (%) on J&WDB‑Select 624 UI for 467Area RSD (%) onJ&W HP‑INNOWaxRT RSD (%) onJ&W .0782.40.034Methylene 039Tetrahydrofuran2.30.029Coelutes with methylenechlorideCoelutes with ene4.40.00564.70.026p-XyleneCoelutes with m-xyleneCoelutes with m-xylene4.40.016o-Xylene4.10.00544.10.31Response (pA)Class 2B solvents on time (min)11121314151617Figure 6. Chromatogram of the USP residual solvents class 2B standard solution resolved on a J&W DB-Select 624 UI for 467 GC column.7
4.7958.6796.73212.123Response (pA)80151237344526768910Retention time (min)11121314151617Figure 7. Chromatogram of the USP residual solvents class 2B standard solution resolved on a J&W HP-INNOWax GC column.ConclusionThe 8890 GC system, equipped with a7697A headspace sampler and inerttee, provides an excellent method forseparating, identifying, and quantifying allof the relevant residual solvents outlinedby USP 467 . Beyond the expectedcoelutions, the peaks in all three classesare well resolved from each other,showing sufficient S/Ns, and can bequantified repeatably.Reference1.USP 32-NF 27, General Chapter USP 467 Organic volatile impurities,United States Pharmacopeia.Pharmacopoeia Convention Inc.,Rockville, MD, USA.www.agilent.com/chemThis information is subject to change without notice. Agilent Technologies, Inc. 2019Printed in the USA, May 16, 20195994-0442ENTable 5. Repeatability (n 10) for class 2B residual solvents obtained on the J&W DB-Select 624 UI for467 and J&W HP-INNOWax columns.Area RSD (%) onJ&W DB-Select624 UI for 467RT RSD (%) onJ&W DB-Select624 UI for 467Area RSD (%) onJ&W HP-INNOWaxRT RSD (%) onJ&W .80.015Tetralin3.70.00523.80.085Compound
Three stock solutions of residual solvents in DMSO were used: Residual Solvent Revised Method 467 Class 1 (p/n 5190-0490) Residual Solvent Revised Method 467 Class 2A (p/n 5190-0492) Residual Solvent Revised Method 467 Class 2B (p/n 5190-0491) The sample preparation procedures for each of the three classes are listed below:
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