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Determination of Maltitol, Isomaltitol, and Lactitol by High-Ph Anion-Exchange Chromatography with Pulsed Amperometric Detection

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Determination of Maltitol, Isomaltitol, and Lactitol by High-Ph Anion-Exchange Chromatography with Pulsed Amperometric Detection J. Agric. Food Chem. 1999, 47, 157−163 157 Determination of Maltitol, Isomaltitol, and Lactitol by High-pH Anion-Exchange Chromatography with Pulsed Amperometric Detection Tommaso R. I. Cataldi,* Cristiana Campa, Innocenzo G. Casella, and Sabino A. Bufo‡ Dipartimento di Chimica, Universita` degli Studi della Basilicata, Via N. Sauro, 85-85100 Potenza, Italy Disaccharide alditols (DAs) such as maltitol, isomaltitol, and lactitol are increasingly being employed in food industry by virtue of their low hydroscopicity, high stability, and good bulking properties. Still, these compounds are reduced-calorie sweeteners, so they are successfully employed in many dietetic foods, like candies, chocolates, baked products, ice creams, and beverages. Here we describe the determination of maltitol, isomaltitol, and lactitol, along with other common carbohydrates, in some foodstuffs such as toffees, biscuits, creams, sponge cakes, chocolates, roasted malt, and chicory leaves. Separations were accomplished by high-pH anion-exchange chromatography (HPAEC) with pulsed amperometric detection using 40 mM NaOH + 1 mM Ba(CH3COO)2 as the mobile phase. The optimal detection potential (EDET )+0.10 V) was established in voltammetric experiments carried out in batch and flowing stream solutions. Under optimized conditions there was no need for both postcolumn addition of strong bases to the eluent and, even more important, column regeneration between runs. A pellicular column with a relatively low ion-exchange capacity was adopted, which allows a rapid separation of sorbitol, isomaltitol, lactitol, maltitol, glucose, fructose, sucrose, and lactose. The presence in the alkaline mobile phase of barium ions improved selectivity and reproducibility besides shorter analysis times as well. Limits of detection were on the order of 10-20 pmol injected. The contents of DAs and other free sugars in some dietetic foods were evaluated by calibration graphs. Keywords: Alditols; chromatography; amperometric detection; dietetic foodstuffs INTRODUCTION Table 1. Structures of Disaccharide Alditols Studied Traditional synthetic sweeteners such as saccharin and aspartame provide a very low amount of calories and, at the same time, have a sweetening power 200- 300 times higher than that of sucrose. Despite these features, they possess only limited bulking properties and cannot be largely employed in the confectionery industries. Disaccharide alditols (DAs), also known as disaccharide sugar alcohols, are the reduced forms of the corresponding disaccharide compounds (Table 1); they are characterized by lower caloric values but similar sweetening power compared to that of sucrose. Maltitol, 4-O-R-D-glucopyranosyl-D-sorbitol, exhibits re- duced laxative effects but similar high tolerance for humidity as D-mannitol, making it largely preferred in many dietetic products (Rapaille et al., 1995). Isomalt or palatinit is a white and crystalline mixture prepared from sucrose, which is composed of 6-O-R-D-glucopyra- nosyl-D-sorbitol, also known as isomaltitol (GPS), and 6-O-R-D-glucopyranosyl-D-mannitol (GPM). Isomalt has only 45-65% of sucrose’s sweetening power and is then often used in combination with other sweeteners (Willi- bald-Ettle and Schiweck, 1996). The even milder sweet- ening power of lactitol (4-O-â-D-galactopyranosyl-D- sorbitol), which is equivalent to ca. 40%, makes it a very Together with traditional sweeteners or other alditols, versatile ingredient in many low-calorie products (Grim- it is especially employed for confectionery manufactures ble et al., 1988; Van Velthuijsen and Blankers, 1991). (Blankers, 1995; Armstrong, 1998). Conceivably, these compounds are provoking a growing interest in the food * To whom correspondence should be addressed (fax, +39- industry because they are characterized by low hydro- 0971-474223; e-mail, [email protected]). scopicity, high stability, and good bulking properties ‡ Dipartimento di Produzione Vegetale. (McNutt et al., 1996; Koutsou et al., 1996). Indeed, DAs 10.1021/jf9807059 CCC: $18.00 © 1999 American Chemical Society Published on Web 12/08/1998 158 J. Agric. Food Chem., Vol. 47, No. 1, 1999 Cataldi et al. are easily incorporated into processed foods and bever- Voltammetric Apparatus. Voltammetric data were ages and successfully used in many reduced-calorie obtained at a rotating disk electrode (RDE) using a products, such as candies, lollipops, chocolates, cough model EDI101 rotator from Radiometer Analytical drops, baked products, and ice creams. Additionally, (Copenhagen, Denmark) and a model 263A potentiostat such compounds do not promote dental caries probably (EG&G Princeton Applied Research, Princeton, NJ). because they are not fermented significantly in the Data acquisition and potentiostat control were ac- mouth (Grenby et al., 1989). The wide variety of complished with a computer running the M270 electro- foodstuffs in which DAs can be used is particularly chemical research software (EG&G) version 4.11. Elec- helpful for diabetics, because alditols induce a reduced trochemical experiments were made using a conventional increase of insulin and glucose in the blood. single-compartment glass cell (PAR). A gold disk elec- The weak acidic properties of carbohydrates allow trode (2.0-mm diameter; Radiometer Analytical) was their chromatographic separation through ion-exchange used for all voltammetric investigations, and the gold mechanisms. Indeed, high-pH or high-performance anion- surface was polished with 0.05 µm of alumina oxide exchange chromatography coupled with pulsed ampero- powder on microcloth using water as the lubricant prior metric detection (HPAEC-PAD) is a very powerful tool to each series of experiments. A Pt auxiliary electrode for the determination of sugars and alditols (Johnson andaAg|AgCl (saturated KCl) reference electrode were and LaCourse, 1990, 1995; Lee, 1996). Sensitivity, used. High-purity N2 gas was bubbled through the cell selectivity, and reliability of this technique are relevant to remove dissolved O2 prior to scan sweeps, and an aspects for the analysis of food products, often charac- atmosphere of N2 was maintained over the cell solutions terized by complex matrixes. Sugar alcohols owing to during voltammetric experiments. The current-poten- their even weaker acidity are generally well-separated tial (i-E) curves reported correspond to the stable with a macroporous anion-exchange column using highly profiles obtained after four potential cycles. All experi- basic mobile phases, that is, 0.4-0.6 M NaOH (Dionex, ments were carried out at room temperature, ∼20 °C. 1994a,b; Corradini et al., 1997; Cataldi et al., 1997a). HPAEC-PAD System. Carbohydrates analyses were Interestingly, in this work a pellicular anion-exchange performed using a Dionex system (Dionex Corp., Sunny- column, characterized by a relatively lower ion-exchange vale, CA) composed of a metal-free isocratic pump model capacity (Dionex, 1995), was successfully employed for IP20 with on-line degas, a pulsed amperometric detector the separation of some relevant alditols and sugars. (model ED40), and a metal-free rotary injection valve Good resolution of commonly occurring carbohydrates equipped with an injection loop of 10 µL. A Dionex in dietetic foodstuffs was obtained upon optimization CarboPac PA100 analytical column (250-mm × 4-mm of the alkaline eluent composition. Efficiency and re- i.d.) preceded by a guard CarboPac PA100 column (50- producibility of the chromatographic results were sig- mm × 4-mm i.d.) was adopted for the optimized separa- nificantly improved by addition of barium ions to the tions. Such a column is composed by sulfonated eth- mobile phase as has been previously described (Cataldi ylvinylbenzene-divinylbenzene particles agglomerated et al., 1997b). We determined the linearity, sensitivity, with 350 µm of Micro Bead quaternary amine-function- reproducibility of response, and limits of detection. The alized latex. Alternatively, a Dionex CarboPac MA1 applicability of the method has been demonstrated by column, 8.5-µm bead diameter (250-mm × 4-mm i.d.), the analysis of some typical dietetic food products, where coupled with a guard CarboPac MA1 column (50-mm × maltitol, isomalt, and lactitol are used as sweeteners. 4-mm i.d.) was used. The flow-through detection cell (Dionex) is made from a 1.0-mm diameter gold working - | MATERIALS AND METHODS electrode and a pH Ag AgCl combination reference electrode; the titanium body of the cell served as the Chemicals. Sodium hydroxide, 50% solution in water counter electrode. Sodium hydroxide eluents were kept (1.515 g/mL), Ba(CH3COO)2 (99%), R-D-lactose mono- in plastic bottles, and a nitrogen headspace was main- hydrate (97%), D-sorbitol (99+%), D-xylose (99%), lactitol tained on the solutions with a Dionex eluent organizer monohydrate (98%), and MgSO4 (99%) were purchased (EO1). A personal computer equipped with the Kontron from Aldrich Chemical Co. (St. Louis, MO), maltitol 98% PC Integration Pack software (Kontron Instruments, was from Fluka Chemie (Buchs, CH), NaN3, R-D-glucose Milan, Italy) allowed acquisition and processing of (99.5%), sucrose (>99.5%), D-fructose (99%), maltose chromatograms. Unless stated otherwise, the pulsed monohydrate (99%), and isomaltitol (98%) were from amperometric detector settings were the following: EDET )+ ) ) )+ Sigma Chemical Co. (Steinheim, Germany). Other chemi- 0.10 V (tDET 440 ms, tINT 240 ms), EOX 0.80 ) )- ) cals were purchased from Carlo Erba (Milan, Italy) and V(tOX 180 ms), ERED 0.80 V (tRED 360 ms); and
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