Mushroom Tissue2based Flow2injection Fluorescence System for the Determination of Noradrenaline and L2Dopa

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1 ACTA CHIMICA SINICA Vol No a Ξ a Ξ a b a a ( a ) ( b ) g ml g ml g ml g ml - 1 (3 S/ k) 14 d g ml g (R S D ) 3 % ( n = 11) ml - 1 Mushroom Tissue2based Flow2injection Fluorescence System for the Determination of Noradrenaline and L2Dopa WANG Zhou2Ping a ZHANG Zhu2Jun Ξ a FU Zhi2Feng a FANG Lu2Qiu b CHEN Ding2Long a ZHANG Xiao a ( a Department of Chemistry Analytical Science Institute Southwest China Normal University Chongqing ) ( b Department of Chemistry Fuling Teacher s College Chongqing ) Abstract It was found that noradrenaline and L2dopa could be oxidized by soluble oxygen under the catalysis of polyphenol oxidase ( PPO) in mushroom tissue column to produce corresponding tri2 or di2hydroxyindole derivatives in basic condition which can emit strong fluorescence radiation Based on it a novel simple and reagentless mushroom tissue2based fluorescence system combined with flow injection analysis for the determination of noradrenaline and L2 dopa is proposed The fluorescent intensity is linear with the concentrations of noradrenaline in the range of g ml - 1 and L2dopa in the range of g ml - 1 respectively The detection limits in turn are g ml - 1 (3 S/ k) and g ml - 1 (3 S/ k) with relative standard deviations of less than 3 % ( n = 11) The reactor was stable for about 14 d and a complete analysis including sampling and washing could be performed in 1 5 min Various interferences from common ions and antioxidants were examined carefully The proposed system has been applied to the determination of studied substances in pharmaceutical preparations and results are in accordance with those values by official methods The experimental results demonstrated the feasibility and reliability of the proposed method Keywords fluorescence flow injection mushroom tissue noradrenaline L2dopa [1] Ξ E2mail : com ; com ; Tel & Fax : Received May ; revised September ; accepted November (No )

2 No 5 : ) ( ) [2 4] ( ) 1 2 [5 9] - HL22 ( ) F24500 ( Hitachi ) 1 PTFE (0 8 mm i d ) [10] [11 13] [14 15] [16 18] [19] [20 22] ( ) Figure 1 Schematic diagram of the fluorescence flow tissue2based personal computer 1 3 ; g ml mol L 21 (ph g ml ) 7 cm 0 8 cm ( ) g ml g ml - 1 (3 S/ k) 30 min (4 ) mol L - 1 (ph 6 5) 10 ( 30 mg) 100 ml 0 01 mol L - 1 (ph 6 5) 10 ml 100 ml 1 1 biosensor for the determination of noradrenaline and L2dopa P1 P2 peristaltic pump ; V valve ; T tissue reactor ; D detector ; F quartz flow cell ; F24500 F24500 fluorescence spectrophotometer ; Ex Excitation ; Em Emission ; PC (Agaricus bisporus linn ) ( [23 24] ) ( g ml - 1 ) ( ( 2) ) ( g ml - 1 ) [ 2 (4) ] [20 25] 100 ml (4 402 nm 495 nm ) 0 01 mol L - 1 (ph 6 5) 3 mol L - 1 NaOH( ) Na 2 HPO 4 2 [ 2 (4) ] [22] 354 nm NaH 2 PO 4 ( ) (0 01 mol L - 1 ph 6 5) 485 nm (

3 510 Vol (Noradrenaline) (L2dopa) Noradrenaline ; L2dopa ; PPO Figure 2 Schematic representation of the catalytic oxidation of noradrenaline and L2dopa by PPO of mushroom tissue column 1 10 min NaOH P1 (0 5 ml min - 1 ) P2 (3 0 ml min - 1 ) 50 L (NaOH ) 402 nm( ) 354 nm( ) 495 nm( ) 485 nm( ) 10 nm (a) (b) [ ] 2 Figure 3 Fluorescence spectra of the products of noradrenaline (a) and 402 nm L2dopa (b) under the catalysis of polyphenol oxidase in mushroom tissue 495 nm( 3a) 354 nm 485 nm( 3b) NaOH ( 0 3 ) ph 6 5 ( 4) : ml min ml min NaOH ( mol L - 1 ) mol L - 1 NaOH NaOH 3 0 mol L (23 25 ) mol L ph ph ph [ 0 01 mol L - 1 ( ) ( ) (9 0) ] 5 ph 6 5

4 No 5 : ph 50 L [23 24] 0 01 mol L - 1 (ph 6 5) ; 50 L 50 L g ml g ml g ml g ml ( RSD) 2 3 % 2 8 % ; IUPAC 4 (3 S/ k) g ml Figure 4 Effect of temperature on the fluorescence intensity 10-8 g ml ph Figure 5 Effect of ph value on the fluorescence intensity Figure 6 Typical recorder output for different concentrations of noradrenaline made with the mushroom tissue2based flow injection fluorescence system g ml - 1 ; g ml - 1 ; g ml - 1 ; NaOH ml min - 1 : g ml ml min ml min mol L - 1 ; buffer and sample solution ph 6 5 ; temperature d 90 % L Analyte Nordrenaline L2dopa 1 Table 1 Parameters of calibration graphs for determination of noradrenaline and L2dopa Linear range/ (g ml - 1 ) Regression equation Correlation coefficient n HV/ V I = C I = C I = C I = C I = C

5 512 Vol [25] ( 3) 2 3 [23 24] ( 4) 3 ; g ml % 2 14 d Table 2 Tolerable ratios of foreign compounds for g ml - 1 noradrenaline and L2dopa Interferent Tolerable ratio Interferent Tolerable ratio Ascorbic acid > 100 MgSO 4 > 100 Uric acid 100 NH 4 Cl 10 Urea 100 NaCl 100 Glucose 200 Na 2 SO 3 50 Lactate 50 Na 2 CO Oxalate 100 KI 50 2cyclodextrin 100 CuSO Starch 1000 FeCl Sodium bitartrate 10 Ca (NO 3 ) Sucrose 500 Adrenaline 1 Fructose Table 3 Results of the determination of nordrenaline and L2dopa in commercial formulations mg ml mg ml - 1 (mg ml - 1 or mg) a 0 49 ( 1 4 %) 0 49 ( 2 2 %) 0 48 ( 1 7 %) 0 49 ( 1 2 %) mg 9 7 ( 2 4 %) 9 8 ( 1 8 %) a Values are mean of three replicates ( R S D ) mg 9 9 ( 2 1 %) 9 9 ( 2 0 %) 4 Table 4 Results of addition2recovery experiments using nordrenaline and L2dopa with three different standard concentrations Sample Initially presented/ ( g ml - 1 ) Added/ ( g ml - 1 ) Found/ ( g ml - 1 ) Recovery/ % ( n = 3)

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