32 5 206 5 Vol.32,No.5 May2 0 6 DOI:0.3652/j.issn.003-5788.206.05.02 HPLC OptimizationonHPLCdetectionandpre-treatment conditionsofpatulininfreshapple SUN Li-hua CHEN Ji-luan 2,3 QIAN Hui-ping LI Zhi-wen 2,3 4 2,3 PANG Xue-qun 4 YAN Shi-jie 2,3 (., 8320032.,300384 3.,3003844. 50642) (.Food ColegeShihezi University,Shihezi,Xinjiang832003,China 2.Colegeof Food Scienceand BiologicalEngineering,Tianjin Agricultural University,Tianjin300384,China 3.Tianjin Engineering and Technology Research Centerof AgriculturalProducts Procesing,Tianjin300384,China 4.Colegeof Biotechnology,South China Agricultural University,Guangzhou,Guangdong50642,China) : rateforhplc whichalwaysafectthesensitivityandanalysisefi-, HPLC ciencyofthemethodwereoptimizedsystemicaly.theresultsindica- tedthatsolidphaseextraction(spe)techniquesusingfunctionalized : polystyrene-divinylbenzenecolumn (PEP)exhibitedsatisfactoryre- coveriescomparingwithotherextraction methods.undertheopti- =0 90, 0.8 ml/min HPLC, mizedchromatographicconditions,patulincouldbe welseparated fromotherimpuritieswithin4 min,anddetectedwithawidelinear, (8.9%), rangebetween0-25000μg/landadetectionlimitof0μg/l (S/ PEP-SPE N=3)usingacetonitrile/water(90:0,v/v)asmobilephase,0.8 (82.42%~95.57%) (3.26% ~4.4%), ml/minasflowrate.theproposedpep-spe-hplc methodwas, PEP-SPE furtherappliedtothedeterminationofpatulininfreshapplesinthe HPLC rangeof82.42%-95.57%,indicatingitmightserveasaneficient toolfortheaccuratequantificationofpatulininfoodsamples. Keywords:high performanceliquid chromatographysolid-phase : Abstract:Arapidandsensitivemethodwasestablishedfortheaccu- ratedeterminationofpatulininfreshapples.experimentalparame- tersincludingextractionmethods,mobilephasecompositionandflow, : ( :205BAD6B07) ( : 5JCYBJC5300),, :,, : (97-),,, E-mail:yanshijie@26.com :206 0 3 patulinfreshapple [], 53
206 5 (5μm,4.6mm 50 mm),, 276nm, 20 μl,, 30,, 0.8mL/min, ( 2080,585,090) (patulin) (2) : AltimaTM C8 (5μm, 4.6mm 50mm),, 276nm, 20μL 30, [2], 090, (.0,0.8, 0.6mL/min) [3],.2.2 5.0 mg 50μg/kg [4] [9], 50000μg/L SN/T2008 2007, 0,50,, 00,250,500,000,5000,0000,25000μg/L, 0 [5-6],.2.3,, 0.0g, ml000μg/l (SPE HPLC) A [7] ( A 25 ml, 25 Mycrosep 228AflaPat 40kHz 300 W 30 min) 78%~7%,RSD 2.~8.2%, ( A 3 ml, 2h, 2mL,, 25 ml, ) ( Wu [8] A 25mL,, ) ( (), 80.7% ~ 25 ml,, ), 8.45%, 8000r/min 5min, HPLC [0], HPLC.2.4,, PEP () (LLE): [0], :..2.3 4 2.5 ml,.. 5mL 0mL, min :85( 85 mm),, ( 0 ml),, 3 : 98%, TRC, 5 ml : 2600PG/mL, 5mL, :, Fisher, 5g :,, 5 ml 2..2 3, 40 :LC-200,,mL,0.45μm :RE-2000 ml HPLC :Visiprep TM SPE-2, Supelco (2)PEP (PEP-SPE): 2 ml :KQ-500DE,2mL,,.2.3 4 2.5mL, :NY3022L, Origin Membranes.2.2. 54, 00 ml [] ml ml [2] ml, ml () : AltimaTM C8 ml,
32 5 : HPLC,, ml 090,0.45μm ml 2..2, HPLC.2.5,,, 0.0g, 50,00,000μg/L3, 0.8 ml/min, ml,.2.4 2 6.372min 3, 2..3 0~25000μg/L ( 3) 2 2 :, 2. Y =0.897 2X +4.49, R = 2.. 0.99998(n=9) 000μg/L,, 6, 0.53RSD 0.072%, [3] : 3, 0μg/L [4] Agilent00ODS 20% 0% 3.850~6.372 min, (5μm,50mm 4.6mm ), 20 595, ml/min,zaied 80,585 [5] SpherisorbODI, 090, ml/min Figure Efectofdiferentmobilephase 2 ratioonseparation Figure2 Efectofdiferentflowratioonseparation 55
206 5 3 Figure3 Standardcurveofpatulin 2.2 4 : 8.90% 65.38% (P>0.05), (P<0.05), [6], 5 HPLC 65.37% OH H, Figure5 Efect of diferent purification techniques on, VA HPLCseparationforpatulininfreshapple VC, [7]2-22 [8] 2.3.2 [7]23-24 :LLE PEP-SPE PEP-,, SPE 82.42%~95.57%,,,, LLE 78.54% ~80.73%,PEP-SPE 3 (P<0.05) 2.3 LLE PEP-SPE LLE, 2.3. HPLC 5 :PEP-SPEC 2.3.3 2 :PEP-SPE LLE,,,PEP-SPE 6.372min LLE 7.090 min /7, [9], 96mL, 56 PEP 82.42% ~95.57%,,Beltran [20] LLE 7%~75%,Gaspar [2] LLE PEP-SPE Table LLEandPEP-SPErecoveriesandcoeficient ofvariation / LLE PEP-SPE ( μ g L - ) /% /% /% /% 50 80.73 a 6.30 95.57 b 4.34 4 00 78.54 a 5.74 83.65 b 3.26 Figure4 Efectofdiferentextractionmethodson 000 79.35 a 6.58 82.42 b 4.4 recoveries
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