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Analysis of Ascorbic Acid and Isoascorbic Acid in Orange and Guava Fruit Juices Distributed in Thailand by LC-IT-MS/MS

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Abstract

This work describes the determination of ascorbic acid (AA) and isoascorbic acid (IAA) in fruit juices by liquid chromatography coupled with ion trap tandem mass spectrometry (LC-IT-MS/MS), with a reversed phase column (C18) and simple isocratic conditions of 0.1 % formic acid. The negative ion mode of electrospray ionization (ESI) and the MS/MS transition of m/z 175 were used for the quantitation of AA- and IAA-generated product ions with m/z 115. The method was validated in terms of linearity, limit of detection (LOD), limit of quantitation (LOQ), precision, and accuracy. Good linearity was achieved with 0.6 and 1.8 μg/mL for the LOD and LOQ, respectively. The intraday and intermediate precisions were approximately 4 and 7 %, respectively. Recovery percentages ranged from 88 to 108 %. All validation parameters were found to be within the acceptable range for both AA and IAA. Hence, the proposed method is suitable for simultaneously determining AA and IAA. Fourteen fruit juice samples were analyzed including fruit juices from supermarkets, local markets, and laboratory preparations. Ten fruit juice samples (100 %) from different brands in the supermarkets were investigated for AA and IAA content. AA could be detected in all the samples, ranging from 5.2 ± 2 to 44.4 ± 1.3 μg/mL, and the values of vitamin C in 4 guava samples were less than the values specified by the manufacturer. IAA was observed in 4 of 10 samples ranging from 3.4 ± 0.9 to 78.3 ± 3.9 μg/mL, and the highest value of IAA was approximately two fold higher than the highest value of AA. For freshly prepared fruit juices, AA was detected in both local market- and laboratory-prepared juices. The value ranged from 13.2 ± 0.9 to 105.1 ± 1.3 μg/mL. In addition, the AA stability after opening the package was evaluated. The results showed that after 4 days of storage in the refrigerator, approximately 30 and 15 % losses of AA were observed for the orange and guava juices, respectively. Fresh fruit juices were thus demonstrated to be good sources of AA, with the highest value observed for guava juice prepared in the laboratory.

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References

  • Achinewhu SC, Hart AD (1994) Effect of processing and storage on the ascorbic acid (vitamin C) content of some pineapple varieties grown in the Rivers State of Nigeria. Plant Foods Hum Nutr 46:335–337

    Article  CAS  Google Scholar 

  • AOAC (2000a) Method 967.21 for ascorbic acid in vitamin preparations and juices. Official methods of analysis of AOAC International/Dr. Willian Horwitz, 17th edn. AOAC International, Maryland

    Google Scholar 

  • AOAC (2000b) Official method 984.26 for vitamin C in foods. Official methods of analysis of AOAC International/Dr. Willian Horwitz, 17th edn. AOAC International, Maryland

    Google Scholar 

  • Arya SP, Mahajan M, Jain P (2000) Non-spectrophotometric methods for the determination of vitamin C. Anal Chim Acta 417:1–14

    Article  CAS  Google Scholar 

  • Barnes MJ (1975) Function of ascorbic acid in collagen metabolism. Ann N Y Acad Sci 258:264–277

    Article  CAS  Google Scholar 

  • Barros AIRNA, Silva AP, Gonçalves B, Nunes FM (2010) A fast, simple, and reliable hydrophilic interaction liquid chromatography method for the determination of ascorbic and isoascorbic acids. Anal Bioanal Chem 396:1863–1875

    Article  CAS  Google Scholar 

  • Behrens WA, Madere R (1994) Ascorbic acid, isoascorbic acid, dehydroascorbic acid, and dehydroisoascorbic acid in selected food products. J Food Compos Anal 7:158–170

    Article  CAS  Google Scholar 

  • Betz JM, Brown PN, Roman MC (2011) Accuracy, precision, and reliability of chemical measurements in natural products research. Fitoterapia 82:44–52

    Article  CAS  Google Scholar 

  • Breithaup DE (2001) Determination of folic acid by ion-pair RP-HPLC in vitamin-fortified fruit juices after solid-phase extraction. Food Chem 74:521–525

    Article  Google Scholar 

  • Christen WG, Glynn RJ, Sesso HD, Kurth T, MacFadyen J, Bubes V, Buring JE, Manson JE, Gaziano JM (2010) Age-related cataract in a randomized trial of vitamins E and C in men. Arch Ophthalmol 128:1397–1405

    Article  CAS  Google Scholar 

  • Combs GF Jr (2012) The vitamins: fundamental aspects in nutrition and health, 4th edn. AP Academic, California, pp 233–259

    Book  Google Scholar 

  • Deutsch JC (1998) Spontaneous hydrolysis and dehydration of dehydroascorbic acid in aqueous solution. Anal Biochem 260:223–229

    Article  CAS  Google Scholar 

  • Deutsch JC (2000) Dehydroascorbic acid. J Chromatogr A 881:299–307

    Article  CAS  Google Scholar 

  • Drivelos S, Dasenaki ME, Thomaidis NS (2010) Determination of isoascorbic acid in fish tissue by hydrophilic interaction liquid chromatography-ultraviolet detection. Anal Bioanal Chem 397:2199–2210

    Article  CAS  Google Scholar 

  • Duarte TL, Lunec J (2005) Part of the series: from dietary antioxidants to regulators in cellular signalling and gene expression—review: when is an antioxidant not an antioxidant? A review of novel actions and reactions of vitamin C. Free Radic Res 39:671–686

    Article  CAS  Google Scholar 

  • EI (2014) Euromonitor International, Juice in Thailand (country report). http://www.euromonitor.com/juice-in-thailand/report. Accessed 16 Mar 2015

  • Eitenmillar RR, Landen WO (2000) Vitamin analysis for the health and food sciences. CRC, Boca Raton, pp 223–270

    Google Scholar 

  • Fabianek J, Hepr H (1967) Antiscorbutic activity of D-araboascorbic acid. Proc Soc Exp Biol Med 125:462–465

    Article  CAS  Google Scholar 

  • FDA (1998) Food and Drug Administration, Thai recommended daily intakes (Thai RDI). http://newsser.fda.moph.go.th/food/file/Laws/Notification%20of%20Ministry%20of%20PublicHealth/Law03P182.pdf. Accessed 30 Mar 2015

  • FDA (2005) Food and Drug Administration, Food additive recommendation in Thailand. http://www.ratchakitcha.soc.go.th/DATA/PDF/0E/00153091.PDF. Accessed 16 Mar 2015

  • Fenoll J, Martínez A, Hellín P, Flores P (2011) Simultaneous determination of ascorbic and dehydroascorbic acids in vegetables and fruits by liquid chromatography with tandem-mass spectrometry. Food Chem 127:340–344

    Article  CAS  Google Scholar 

  • Fontannaz P, Kilinc T, Heudi O (2006) HPLC-UV determination of total vitamin C in a wide range of fortified food products. Food Chem 94:626–631

    Article  CAS  Google Scholar 

  • Furusawa N (2001) Rapid high-performance liquid chromatographic identification/quantification of total vitamin C in fruit drinks. 12:27–29

  • Ghorbani AR, Momenbeik F, Khorasani JH, Amini MK (2004) Simultaneous micellar liquid chromatographic analysis of seven water-soluble vitamins: optimization using super modified simplex. Anal Bioanal Chem 379:439–444

    Article  CAS  Google Scholar 

  • Gutzeit D, Baleanu G, Winterhalter P, Jerz G (2008) Vitamin C content in sea buckthorn berries (Hippophaë rhamnoides L. ssp. rhamnoides) and related products: a kinetic study on storage stability and the determination of processing effects. J Food Sci 73:C615–C620

    Article  CAS  Google Scholar 

  • Hathcock JN, Azzi A, Blumberg J, Bray T, Dickinson A, Frei B et al (2005) Vitamins E and C are safe across a broad range of intakes. Am J Clin Nutr 81:736–745

    CAS  Google Scholar 

  • Hemilä H (2011) The effect of vitamin C on the common cold. J Pharm Pract 24:241–242

    Article  Google Scholar 

  • Honarbakhsh S, Schachter M (2009) Vitamins and cardiovascular disease. Br J Nutr 101:1113–1131

    Article  CAS  Google Scholar 

  • IOM (2014) Institute of Medicine, National Academies, Recommended dietary allowances and adequate intakes: vitamins. http://iom.edu/Activities/Nutrition/SummaryDRIs/~/media/Files/Activity%20Files/Nutrition/DRIs/New%20Material/5DRI%20Values%20SummaryTables%2014.pdf. Accessed 18 Mar 2015

  • Johnston CS, Bowling DL (2002) Stability of ascorbic acid in commercially available orange juices. J Am Diet Assoc 102:525–529

    Article  Google Scholar 

  • Johnston CS, Hale JC (2005) Oxidation of ascorbic acid in stored orange juice is associated with reduced plasma vitamin C concentrations and elevated lipid peroxides. J Am Diet Assoc 105:106–109

    Article  CAS  Google Scholar 

  • Johnston CS, Steinberg FM, Rucker RB (2007) Ascorbic acid. In: Zempleni J, Rucker RB, McCormick DB, Suttie JW (eds) Handbook of vitamins, 4th edn. CRC, Boca Raton, pp 489–520

    Google Scholar 

  • Kabasakalis V, Siopidou D, Moshatou E (2000) Ascorbic acid content of commercial fruit juices and its rate of loss upon storage. Food Chem 73:325–328

    Article  Google Scholar 

  • Kalt W (2005) Effects of production and processing factors on major fruit and vegetable antioxidants. J Food Sci 70:R11–R19

    Article  CAS  Google Scholar 

  • Klejdus B, Petrlová J, Potěšil D, Adam V, Mikelová R, Vacek J, Kizek R, Kubáň V (2004) Simultaneous determination of water- and fat-soluble vitamins in pharmaceutical preparations by high-performance liquid chromatography coupled with diode array detection. Anal Chim Acta 520:57–67

    Article  CAS  Google Scholar 

  • Klimczak I, Gliszczyńska-Świgło A (2015) Comparison of UPLC and HPLC methods for determination of vitamin C. Food Chem 175:100–105

    Article  CAS  Google Scholar 

  • Laur M, Tian L (2011) Provitamin A and vitamin C contents in selected California grown cantaloupe and honeydew melons and imported. J Food Compos Anal 24:194–201

    Article  CAS  Google Scholar 

  • Mandl J, Szarka A, Bánhegyi G (2009) Vitamin C: update on physiology and pharmacology. Br J Pharmacol 157:1097–1110

    Article  CAS  Google Scholar 

  • Masamba KG, Mndalira K (2013) Vitamin C stability in pineapple, guava and baobab juices under different storage conditions using different levels of sodium benzoate and metabisulphite. Afr J Biotechnol 12:186–191

    CAS  Google Scholar 

  • Novakova L, Solich P, Solichova D (2008) HPLC methods for simultaneous determination of ascorbic and dehydroascorbic acids. TrAC Trends Anal Chem 27:942–958

    Article  CAS  Google Scholar 

  • Novakova L, Solichova D, Solich P (2009) Hydrophilic interaction liquid chromatography—charged aerosol detection as a straightforward solution for simultaneous analysis of ascorbic acid and dehydroascorbic acid. J Chromatogr A 1216:4574–4581

    Article  CAS  Google Scholar 

  • Odriozola-Serrano I, Hernandez-Jover T, Martin-Belloso O (2007) Comparative evaluation of UV–HPLC methods and reducing agents to determine vitamin C in fruits. Food Chem 105:1151–1158

    Article  CAS  Google Scholar 

  • Padayatty SJ, Levine M (2001) New insights into the physiology and pharmacology of vitamin C. CMAJ 164:353–355

    CAS  Google Scholar 

  • Pisoschi AM, Danet AF, Kalonowski S (2008) Ascorbic acid determination in commercial fruit juice samples by cyclic voltammetry. J Autom Methods Manage Chem. doi:10.1155/2008/937651

  • Quirós ARB, Fernández-Arias M, López-Hernández J (2009) A screening method for the determination of ascorbic acid in fruit juices and soft drinks. Food Chem 116:509–512

    Article  Google Scholar 

  • Rehwoldt R (1986) Tracking the use of antioxidants through industry surveys. Food Chem Toxicol 24:1039–1041

    Article  CAS  Google Scholar 

  • Ribani M, Bottoli CBG, Collins CH, Jardin ICSF, Melo LFC (2004) Validation for chromatographic and electrophoretic methods. Quim Nov. 27:771–780

  • Russell LF (2004) Water-soluble vitamins. In: Nollet LML (ed) Handbook of food analysis: physical characterization and nutrient analysis. Marcel Dekker, New York, pp 487–571

    Google Scholar 

  • Schell DA, Bode AM (1993) Measurement of ascorbic acid and dehydroascorbic acid in mammalian tissue utilizing HPLC and electrochemical detection. Biomed Chromatogr 7:267–272

  • Scherer R, Rybka ACP, Ballus CA, Meinhart AD, Filho JT, Godoy HT (2012) Validation of a HPLC method for simultaneous determination of main organic acids in fruits and juices. Food Chem 135:150–154

    Article  CAS  Google Scholar 

  • Schlueter AK, Johnston CS (2011) Vitamin C: overview and update. JEBCAM 16:49–57

    CAS  Google Scholar 

  • Shah VP, Midha KK, Dighe S, McGilveray IJ, Skelly JP, Yacobi A et al (1992) Analytical methods validation: bioavailability, bioequivalence and pharmacokinetic studies. Pharm Res 9:588–592

    Article  Google Scholar 

  • Spínola V, Mendes B, Câmara JS, Castilho PC (2012) An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables. Evaluation of degradation rate during storage. Anal Bioanal Chem 403:1049–1058

    Article  Google Scholar 

  • Steskova A, Morochovicova M, Leskova E (2006) Vitamin C degradation during storage of fortified foods. J Food Nutr Res 45:55–61

    CAS  Google Scholar 

  • Taverniers I, Loose MD, Bockstaele EV (2004) Trends in quality in the analytical laboratory. II. Analytical method validation and quality assurance. TrAC Trends Anal Chem 23:535–552

    Article  CAS  Google Scholar 

  • Ullah S, Hussain A, Ali J, Khaliqurrehman, Ullah A (2012) A simple and rapid HPLC method for analysis of vitamin-C in local packed juices of Pakistan. Middle-East J Sci Res 12:1085–1091

    CAS  Google Scholar 

  • Vanderslice JT, Higgs DJ, Hayes JM, Block G (1990) Ascorbic acid and dehydroascorbic acid content of foods-as-eaten. J Food Compos Anal 3:105–118

  • Versari A, Mattioli A, Parpinello GP, Galassi S (2004) Rapid analysis of ascorbic and isoascorbic acids in fruit juice by capillary electrophoresis. Food control 15:355–358

  • Walker R (1991) Erythorbic acid and its sodium salt. In: Joint FAO/WHO Expert Committee on Food Additives (JEFCA) (ed) Toxicological evaluation of certain food additives and contaminants. World Health Organization, Geneva, pp 27–60

    Google Scholar 

  • Weber P, Bendich A, Schalch W (1996) Vitamin C and human health—a review of recent data relevant to human requirements. Int J Vitam Nutr Res 66:19–30

    CAS  Google Scholar 

  • Wilson JX (2002) The physiological role of dehydroascorbic acid. FEBS Lett 527:5–9

    Article  CAS  Google Scholar 

  • Yan B, Cao G, Sun T, Zhao X, Hu X, Yan J, Peng Y, Shi A, Li Y, Xue W, Li M, Li K, Liu Y (2014) Determination of pinocembrin in human plasma by solid-phase extraction and LC/MS/MS: application to pharmacokinetic studies. Biomed Chromatogr 28:1601–1606

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, 73170, Thailand

    Somchai Boonpangrak & Sasiwimon Lalitmanat

  2. Department of Clinical Microscopy, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Yaneenart Suwanwong

  3. Center of Data Mining and Biomedical Informatics, Faculty of Medical Technology, Mahidol University, Bangkok, 10700, Thailand

    Supaluk Prachayasittikul

  4. Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, 73170, Thailand

    Virapong Prachayasittikul

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  1. Somchai Boonpangrak
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Correspondence to Virapong Prachayasittikul.

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Funding

This research project was supported by Mahidol University and by the Office of the Higher Education Commission, Mahidol University, under the National Research Universities Initiative and Annual Government Grant under Mahidol University (2556–2558 B.E.).

Conflict of Interest

Somchai Boonpangrak declares that he has no conflict of interest. Sasiwimon Lalitmanat declares that she has no conflict of interest. Yaneenart Suwanwong declares that she has no conflict of interest. Supaluk Prachayasittikul declares that she has no conflict of interest. Virapong Prachayasittikul declares that he has no conflict of interest.

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Boonpangrak, S., Lalitmanat, S., Suwanwong, Y. et al. Analysis of Ascorbic Acid and Isoascorbic Acid in Orange and Guava Fruit Juices Distributed in Thailand by LC-IT-MS/MS. Food Anal. Methods 9, 1616–1626 (2016). https://doi.org/10.1007/s12161-015-0337-x

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