Validation of a HPLC-DAD method for quantification of ascorbic acid in mixed juice powder of acerola and seriguela / Validação de um método HPLC-DAD para quantificação de ácido ascórbico de suco misto em pó de acerola e seriguela

Christine Maria Carneiro Maranhão Ribeiro, Anne Jaciele da Silva, Clayton A. Azevedo Filho, Emmanuela Prado de Paiva Azevedo, Marcella Melo Assis Costa, Maria Inês Sucupira Maciel


The objective of this work was to optimize extraction process and to validate the HPLC-DAD method for quantification of ascorbic acid content in mixed juice powder of acerola and seriguela with different dextrose equivalent maltodextrin (5, 10, 15) as an encapsulating agent. The influence of the extraction variables was studied through factorial design. Control variables were mass, percent of orthophosphoric acid and stirring time. Chromatographic method was validated in terms of precision, selectivity, linearity and accuracy. Chromatographic and titrimetry results were compared for all dextrose equivalent. Mass and percentage of orthophosphoric acid presented p < 0.05. There were no significant differences among the dextrose equivalents ranging from 2,830.1 mg/100 g to 2,931.61 mg/100 g of ascorbic acid. Both values, intra and intermediary precision test coefficients of variation were less than 5%. The correlation coefficient was 0.99. Respective value of limit of detection was 0.001 mg/mL and for quantification was 0.003 mg/mL. The application of HPLC-DAD method can be considered acceptable for quantification ascorbic acid in fruit powder.


factorial design, validation, hplc-dad, vitamin c, spray drying.

Full Text:



ABE-MATSUMOTO, L. T. et al. Is Titration as Accurate as HPLC for Determination of Vitamin C in Supplements ? — Titration versus HPLC for Vitamin C Analysis. 2020. p. 269–279.

BOONPANGRAK, S. et al. Analysis of Ascorbic Acid and Isoascorbic Acid in Orange and Guava Fruit Juices Distributed in Thailand by LC-IT-MS/MS. Food Analytical Methods, 24 jun. 2016. v. 9, n. 6, p. 1616–1626. Disponível em: .

BRAININA, K. Z. et al. Electrochemical sensor based on a carbon veil modified by phytosynthesized gold nanoparticles for determination of ascorbic acid. Sensors (Switzerland), 2020. v. 20, n. 6.

CUNHA-SANTOS, E. C. E. et al. Vitamin C in camu-camu [Myrciaria dubia (H.B.K.) McVaugh]: evaluation of extraction and analytical methods. Food Research International, 2019. v. 115, n. August 2018, p. 160–166. Disponível em: .

CURI, P. N. et al. Optimization of native Brazilian fruit jelly through desirability-based mixture design. Food Science and Technology, 2019. v. 39, n. 2, p. 388–395.

DEPARTAMENTO DE ALIMENTAÇÃO E NUTRIÇÃO. Controlo da Qualidade Interno Métodos Cromatográficos. [S.l.]: Instituto Nacional de Saúde Doutor Ricardo Jorge, 2012.

GERGORIC, M.; BARRIER, A.; RETEGAN, T. Recovery of Rare-Earth Elements from Neodymium Magnet Waste Using Glycolic, Maleic, and Ascorbic Acids Followed by Solvent Extraction. Journal of Sustainable Metallurgy, 2019. v. 5, n. 1, p. 85–96. Disponível em: .

GROSSO, C. et al. Box–Behnken factorial design to obtain a phenolic-rich extract from the aerial parts of Chelidonium majus L. Talanta, 2014. v. 130, p. 128–136.

IGUAL, M. et al. Optimization of spray drying conditions for lulo (Solanum quitoense L.) pulp. Powder Technology, 2014. v. 256, p. 233–238. Disponível em: .

INFANTE, J. et al. Antioxidant and anti-inflammatory activities of unexplored Brazilian native fruits. Biological Properties of Unexplored Native Fruits syndrome, 2016. v. 11, n. 4, p. 1–13.

INTERNATIONAL CONFERENCE ON HARMONIZATION- ICH. Guidance for industry Q2b: validation of analytical procedures: methodology. [S.l.]: Rockville: US. Federal Register, 1997.

IOM. Food and Nutrition Board. Standing Committee on the Scientific Evaluation of Dietary Reference Intakes. In: PRESS, T. N. A. (Org.). Food and Nutrition Board. Standing Committee on the Scientific Evaluation of Dietary Reference Intakes. Washington: [s.n.], 2010.

ISLAM, M. Z. et al. Effect of vacuum spray drying on the physicochemical properties, water sorption and glass transition phenomenon of orange juice powder. Journal of Food Engineering, jan. 2016. v. 169, p. 131–140. Disponível em: . Acesso em: 21 set. 2015.

KRISHNAIAH, D.; NITHYANANDAM, R.; SARBATLY, R. A critical review on the spray drying of fruit extract: effect of additives on physicochemical properties. Critical reviews in food science and nutrition, 2014. v. 54, n. 4, p. 449–73. Disponível em: .

MARANHÃO RIBEIRO, C. M. C. et al. Fruit development and ripening of acerola ‘Okinawa’ cultivar. Acta Horticulturae, 2018. v. 1198, p. 199–204.

MOREIRA, G. É. G. et al. Ascorbic Acid and Anthocyanin Retention During Spray Drying of Acerola Pomace Extract. Journal of Food Processing and Preservation, 2010. v. 34, n. 5, p. 915–925. Disponível em: .

OFFICIAL METHODS OF ANALYSIS. AOAC INTERNATIONAL. 18th Ed ed. Gaithersburg, MD, USA: [s.n.], 2005.

PATIL, V.; CHAUHAN, A. K.; SINGH, R. P. Optimization of the spray-drying process for developing guava powder using response surface methodology. Powder Technology, fev. 2014. v. 253, p. 230–236. Disponível em: . Acesso em: 31 maio 2014.

PHILLIPS, K. M. et al. Stability of vitamin C in fruit and vegetable homogenates stored at different temperatures. Journal of Food Composition and Analysis, 2016. v. 45, p. 147–162.

PRAKASH, A. et al. Implication of processing and differential blending on quality characteristics in nutritionally enriched ketchup (Nutri-Ketchup) from acerola and tomato. Journal of Food Science and Technology, 2016. v. 53, n. 8, p. 3175–3185. Disponível em: .

PYCIA, K. et al. Maltodextrins from chemically modified starches. Selected physicochemical properties. Carbohydrate polymers, 1 ago. 2016. v. 146, p. 301–9. Disponível em: . Acesso em: 30 abr. 2016.

RIBEIRO, C. M. C. M. et al. Optimization of the spray drying process conditions for acerola and seriguela juice mix. Food Science and Technology, 2019. v. 39, p. 48–55.

______ et al. Hygroscopic behavior of spray dried acerola and seriguela mixed juice powder stored Comportamento higroscópico de suco misto de acerola e seriguela em pó armazenado. 2020. n. 50, p. 109–119.

SHAH, V. P. et al. Analytical Methods Validation : Bioavailability , Bioequivalence , and Pharmacokinetic Studies. Journal of Pharmaceutical Sciences, 1992. v. 81, n. 3, p. 309–312.

SILVA, L. M. R. et al. Development of mixed nectar of cashew apple, mango and acerola. International Food Research Journal, 2017. v. 24, n. 1, p. 232–237.

SILVA, R. V. et al. In vitro photoprotective activity of the Spondias purpurea L. peel crude extract and its incorporation in a pharmaceutical formulation. Industrial Crops and Products, maio. 2016. v. 83, p. 509–514. Disponível em: . Acesso em: 17 fev. 2016.

SOARES, M.; OLIVEIRA, C. Molecular analysis of alternative oxidase and ascorbic acid biosynthesis in two acerola clones (Malpighia emarginata DC). Brazilian Journal of Development, 2021. v. 7, n. 5, p. 48548–48569.

TARRAGO-TRANI, M. T.; PHILLIPS, K. M.; COTTY, M. Matrix-specific method validation for quantitative analysis of vitamin C in diverse foods. Journal of Food Composition and Analysis, 2012. v. 26, n. 1–2, p. 12–25.

TASHKHOURIAN, J.; VALIZADEH, H.; ABBASPOUR, A. Ascorbic Acid Determination Based on Electrocatalytic Behavior of Metal-Organic Framework MIL-101-(Cr) at Modified Carbon-Paste Electrode. Journal of AOAC INTERNATIONAL, 1 mar. 2019. v. 102, n. 2, p. 625–632. Disponível em: . Acesso em: 8 fev. 2021.

VANIN, F. M.; CARVALHO, R. A. De. Vitamin C stability in acerola and camu-camu powder obtained by spray drying. Braz. J. Food Technol., 2020. v. 23, p. 1–13.