ANION CHROMATOGRAPHY WITH POLYSPHER® IC AN-1 MERCKPolyspher® IC AN-1 column was devel- oped for use in ion chromatography for anionic separation. Packed with an ion ‘exchange resin made by bonding a quater- nary ammonium group to hydrophilic gel, the colurnn is best suited for high speed ion chromatography without use of a suppres- sor. SPECIFICATIONS 1.) Column dimension: Main column; Prepacked column RT 100-4.6 Guard column: Prepared column AT 10-4.6 2.) Sorbent: lon exchange resin made by bonding a quaternary ammonium group to hydrophilic polymethacrylate-type gel. 3) Particle size: 12 ym 4.) pH stability: 2-10 5.) Detection: Conductivity or indirect uv. Highly sensitive conductivity detection is possible. 1.0ng (2 ppb) of NOz, 1.0 ng (2 ppb) of NOg or 1.5 ng (3 ppb) of SO.” are the minimum detection limits when 500 pil of sample is injected. MOBILE PHASE Since Polyspher® IC AN-1 has a wide range of usable pH's excellent separations ‘of most inorganic anions is attainable. in general, the mobile phase is an aqueous: solution of phthalic acid, p-hydroxybenzoic acid, tartaric acid, oxalic acid or boric acid. ‘Change from one concentration, pH or kind of the mobile phase to another will change the length of time and the order in. which the anions will be eluted (Figure 18). All the listed eluents can be used for con- ductivity detection, Mobile phases based on phthalic acid or p-hydroxybenzoic acid can be used for conductivity and indirect UV detection (UV = 254 nm). 1.) 1.5 mM phthalic acid + 1.38mM tris(hydroxymethy!) _aminomethane (= TRIS) +300 mM boric acid, pH 4.0: This eluent is best suited to routine analysis of the most important anions H,PO,;, F, Cr, NOz, Br, NOs’, SO. in one single run (Figure 1a, *b and 2) and can be used also for the separa- tion of organic anions such as malate, oxalate, fumarate, citrate, tartrate, acetate, pyruvate or lactate (Figure 3), 2) 2.6mM phthalic acid, pH 4.0 (adjusted with TRIS) Most of the important anions such as F of H,PO,, CI’, NOz’, Br, NOs" and $0,* can be separated in a very short run, Therefore, this eluent is suited to routine analysis (Figure 4, 5, 6). 3.) 2.5 mM phthalic acid, pH 4.2 (adjusted with TRIS): The nine anions F, Cl’, N02 .Br, NOs, S047, $20,*, J, SCN" are separated (Figure 7) in a very short run. 4.) 0.9mM_ sodium 1-octanesulfonate + 20 mM boric acid, pH 4.6: This mixture of octanesultonic and boric acid is used as the eluent for the separation of F, CI, NO, Br’, NOg’, J, SO,4*, $2037 in a single run (Figure 8). This eluent is good for obtaining a low background and a stable baseline even when a large quantity of sample is injected. There- fore, it is suited to low concentration samples (Figure 9). The minimum detection limits with this eluent are 1 ng NOs" (2 ppb NOs"), 1ng NOx (2 apd NOs), 1.5ng SO? (ppb SO.“), when 500 pi of sample is injected. No5) 6) 7) 9) 10.) 0.25 mM p-hydroxybenzoic system peak will appear with this eluent 3mM trimesic acid + 0.04 mM EDTA- Nap, pH 3.3: This eluent is suited to the separation of three kinds of acids, phosphoric acid (H_PO,)), pyrophosphoric acid (HjP,077) and tripolyphosphoric acid (HgP30j5%) in a single run (Figure 11). 1 mM tartaric acid, pH 3.3: Anions that elute before chloride, such as acetic acid, succinic acid, HaPO,", HyPOs, HzPO2, ClOz, BrOs, are separated (Figure 12). 2 mM oxalic acid, pH 2.9: This eluent can be used especially for the analysis of F* and HPO," (Figure 13). 1.5 mM p-hydroxybenzoic acid + 10 % methanol, pH 7.8 (adjusted with N,N- diethylethanolamine = DEEA) This eluent is used for the separation of the standard anions F, CI’, NO;’, Br, NOs, HPO,4?", SO," (Figure 14) in a single run. But special anions such as HASO4*, U”, CrO4?, S203, WO, Mo0,*", SCN” can also be separated. 1.5mM p-hydroxybenzoic acid+ 5 % acetonitrile, pH 8.0 (adjusted with DEEA). The standard anions F, Cl’, NO2”,Br’, NO,", HPO,?", SO," can be separated in a single run. acid + 10 % methanol, pH 8.7 (adjusted with DEEA). This special eluent is suited to the separation of S04”, S03”, S203”, SCN in a single run (Figure 15). / J 11,)2.5 mM prhydroxybenzoic acid + 10% methanol, pH9.7 (adjusted with DEEA): Anions such as SO,*, S,0,? and polythionic acids (S30,%, S40,*, etc.) are separated (Figure 16), 12.) 1 mM KOH + 0.1 mM p-hydroxybenzoic acid: This eluent is suited to the separ of anions having low dissociation con- stants, such as cyanogens and silicic acids (Figure 17). 13.) 4.5 mM boric acid + 13.5 mM manni- tol+ 3.8mM TRIS + 2% acetonitrile, pH 7.85: This eluent can be used for the sepa- ration of the standard anions F, Cr, NOz, Br, NOs", HPO,”, SO,” in a single run CONCENTRATION OF ANIONS The prepacked column RT 10-4.6 Poly- spher® IC AN-1 (Guard column) can be used for the concentrations of arions. The guard column has to be placed in the loop of an injector. The sample has to be injected into the guard column with the injector at "Load" position. The sample anions will thus be concentrated in the guard column. The concentrated sample will be transported into the analytical column with the injector at “inject’ position. Even after injecting large sample volumes with low concentrations of anions, good separations can be obtained (Figure 10). /_ RT 10-4,6 Polyspher IC AN-1 fain sample a lee ump colume‘APPLICATIONS WITH PREPACKED COLUMN RT 100-4,6 POLYSPHER® IC AN-1 Figure 18: Standard anions. (Conductivity detection). ‘Semple: 20 pt 1.HgPOg 10 ppm are 10pm 3 s.ct 10 ppm 4.NOg —— 10pom 5.87 10pm B.Nos iOpen 7.807" 10 ppm S10 tS mn. Sluent: 4.5 mM phthalic acid + 1.88 M TRIS + 300 mM boric acid, pH 4.0 4.3 mimi Conductivity 35°C. Figure 2: Tap water (enriched with 1 ppm anion standard). ‘Sample: 20 1othPOe ar cr 4.NOy 5a 2 7]| Nos: 780% 1048 min 4.5 mM phthalic acid + 1.38 mM TRIS + 300 mM boric acid, pH 4.0 Flow rate: 4.3.mlirin Detection: “Indirect UV (254 nm) Column temp: 35°C. Figure 1b:- Standard anions. (Indirect UV detection). Sarria: 20 A.HgPOy 1 pom 2F° Spm scr ppm 4.Nop —10ppm ser 1Opem BNO 10pm TiL* 7.0, — top—m 23 rr Eluent 4.5 mM phthalic acic + 1.38 mM TRIS + 300 mM boric acid, pH 4.0 Flow rate: 4.3.mUimin Detection: _indiract UV (264 nm Column temp: 35°C Figure 3: Beer. ja Sample: ber uted 100, 20 ji Tr acetic aid 14 9pm 2. HyPO¢, succinic aid rosie aio 3. laewe see, pyruvie ald 4cr 83 pem 5 mall acid 85 pe 6 2 NO 1.1 ppm ovale seid 1.2 99m 8 civicscid 82 pom 2 9.50" ¢3pem 20 fin, Eluent: 41,5. mM phthalic acid + 1.38 mM TRIS + 300 mM boric acid, pH 4.0 : Flow rate 1.2 milimin Detection: Conductivity Column temp: 40°CFigure 4: Standard anions. Semple: 30x 1. H,POX 30 99m Zot 5 pom 3. NO 10.p0m 48° 10p0m 5. NOY 10 ppm 6 So; 10 99m 2.5 mM phthalic acid, pH 4.0 (adjusted with TRIS) 4.5 miimin Detection: Conductivity Column temp.: 40°C Figure 6: Chinese cabbage, a) 1. HyPOC 80pm Zor | 70m 3. NOs” 140,p0m $50 200 pom 1. HyPOe 70 pom 2cr "160 pom 2 3. NOx 300 pom 4.50% 32000 HL Mi é 12 min Eluent: 2.5 mM phthalic seid, pH 4.0 (adjusted with TRIS) Flow rate 1.2. mmin Detection: Conductivity Column temp.; 40°C. ‘Sample: Leaves (A) and staiks (B) of chinese ‘cabbage are analyzed. lons were extracted using ‘a juices-mixer and then the liquid was fiterered. Figure 5: Sea water. cto 028 et wien Eluent 2.5 mM phthalic acid, pH 4.0 (Gdjusted with TRIS) Flowrate; 1.2 m/min Detection: Conductivity Column temp.: 40°C Sampe pre treatm: 800 chapter E.5 Figure 7: Standard anions. seit 0 2 © 8 1alli2 min Eluent 2.5 mM phthalic acti, pH 4.2 (adjusted with TRIS) Flow rate: 1.5 mimin Detection: Conductivity Column temp: 40°CFigure 8: Standard anions. Semple: standard 15 wr 5 pom 201 5 pom 3.80 100m 48 tO ppm S.NOy i0ppm ar 20 ppm 7.802" Y0pom 8 S:05" 10 pom 8 6 é 2 emir Eluent: 0.9 mM sodium 1-octane- sulfonate + 20 mM borie acid, pHAs Flow rate: 1.2.mimin Detect Conductivity Column temp: 40°C Figure 10: Concentration of anions. ‘Semple: Or pure water spike! with TF Sppb 2er Spb 3.NO 10 ppb 4.8 spp 5. NOY 10pe 2 6.500 10p00, 3 , 4s ; f\ ( 6 oo tom Concentrator, RT 10-4,6 Polyspher IC AN-1 0.9 mM sodium t-octanasutfo- hate + 20 mM boric acid, pH 4,6 4.2 mimi Condtuetivity Column temp.: 40°C Figure 9: Pure water. Semple: purified wate, 500 2.3 pp> 2.5 poo 4.808 e908 1 | 23 4 6 12 min nt: ° Bue 0.9 mM sodium 1-octane- sulfonate +20 mM! boric acid Flow rate: 1.2 miimin Detection: Conductivity Column temp.: 40°C Figure 11: Standard anions. Som 1 HaPO." 20 0m 2. HyP:03° 20 com 3. HsPi0%s 50 pom Eluent: 3 mM trimesic acie + 0.04 mM EDTA-Neo, pH 3.3 Flow rate: 1.5 mumin Detection: Conductivity Column temp: 40°CFigure 12: Standard anions. Sample seat acid succinic acid HPO. PO, HP0, Clos, 5105" cr 8 10 12 14 16 18 min, 1 mM tartaric acid, pH 3.3 4.8 m/min Conductiviey Column temp. 40°C Figure 14: Standard anions. 16 min 1.5 mM p-hydronybenzoic acid +10 % methanol, pH7.B (adjusted wit DEEA) 4.2.mmin Conductivty Column temp: 40°C Figure 13: River water. 18) Sample: (8) Sarat: Unpolsted rier water eluted iver water Lee Se Fe 22096 BPO." Sopp 2 myPO" 980pp 204 om 2 2 2 ° 2 Eluent: 2 mM oxalic acid, pH 2.9 Fiow rate: 4.5 mlimin Detection: Conductivity Column temp. 40°C Figure 1: Standard anions. ‘f 3 DUK Sample: 1001 Oz" S ppm S037 10 ppm 5,03" 10p9m SCN” 10pom 2 20min, 0.25 mM p-hydroxybenzoie acid + 10% methancl, pH 9.7 (adjusted with DEA) 4.2 mimi Conductivity 50°CFigure 16: Standard anions. : Sample: 50.1 150," 100m 2. $:0,3° 10ppm 2. 8,0¢" 4000 48.04" 80 ppm : A ul pray Etuent 2.5 mM p-hydroxybenzoie acid + 10% methanol, pH 8.7 (adjusted with DEA) Flow rate: 1.2 mimi Detection: Conductivity Column temp.: 40°C Figure 18: influence of pH on retention time. 2H of motile sham 2.5 mM phthalic acid, PH variable 1.5 m/min Conductivity Column teme.: 40°C: Event: Flow rate: Detection: Column temp. : Standard anions. Semole: 40 1. WSO. 590m cr ppm 2.0N" 10pm 4.8" 10pem 5. NOs" 10pm 6.50, 10 9pm prhyorony benzole eid “ 1 mM KOH + 0.1 mM, Pehydroxybenzois acid 1.5 mimin Conductivity 45°C