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Development and validation of a RP-HPLC method for a simultaneous analysis of quercetin and ascorbic acid in psidium guajava fruit extract at different ripening stages
Sulastri A.a,b, Maulana Y.E.c, Amaliya A.d, Sukrasno S.a, Soemardji A.A.a
a Pharmacology-Clinical Pharmacy Research Group, Institut Teknologi Bandung, Bandung, 40132, Indonesia
b Faculty of Sport and Health Education, Universitas Pendidikan Indonesia, Bandung, 40154, Indonesia
c Integrated Laboratory, Politeknik Kesehatan Bandung, Cimahi, 40514, Indonesia
d Department of Periodontology, Faculty of Dentistry, Universitas Padjadjaran, Bandung, 40132, Indonesia
[vc_row][vc_column][vc_row_inner][vc_column_inner][vc_separator css=”.vc_custom_1624529070653{padding-top: 30px !important;padding-bottom: 30px !important;}”][/vc_column_inner][/vc_row_inner][vc_row_inner layout=”boxed”][vc_column_inner width=”3/4″ css=”.vc_custom_1624695412187{border-right-width: 1px !important;border-right-color: #dddddd !important;border-right-style: solid !important;border-radius: 1px !important;}”][vc_empty_space][megatron_heading title=”Abstract” size=”size-sm” text_align=”text-left”][vc_column_text]© 2020 Taylor’s University. All rights reserved.A sensitive yet simple but HPLC-UV method is developed to determine quercetin and ascorbic acid in red guava (Psidium guajava Linn.). The flow rate at 1.0 ml/ min, with an injection of 20 µl, was detected using two different wavelengths; 254 nm for ascorbic acid and 370 nm for quercetin. The eluent system consisted of 0.01 mol/ L NH4H2PO4 with pH = 2.67 (mobile phase A) and a 0.1% orthophosphoric acid mixture: methanol 51:49 (mobile phase B). The gradient elution method used is 0-5 minutes: A 100%; 5-45 minutes A 0%; and 45-55 minutes A 100%. The stationary phase is a column C-18, 150 x 4.6 mm id, 5 µm ODS-3 Inertsil GL-Science, and uses a Shimadzu, LC-20AT prominence. The RT for ascorbic acid was 3.5 minutes, and the RT for quercetin was 25.32 minutes, with the overall analysis duration of 55 minutes. It was obtained, from the validation results, that a linear calibration curve with R = 0.999 for quercetin and R = 0.999 for ascorbic acid, respectively. The precision (RSD) is less than 1.35% for quercetin and 1.65% for Ascorbic acid. The limit of detection (LOD) of the two compounds is 0.01 μg / ml. The limit of quantification (LOQ) of the two compounds is 0.02 μg / ml for quercetin and 0.04 μg/ ml for ascorbic acid. The recovery rates for quercetin were 86.77% and 95.75% for ascorbic acid. The specificity value for ascorbic was 2.59 and for quercetin 2.28. Thus, the HPLC analysis method developed has met the analytical requirements and can be used in the analysis of quercetin and ascorbic acid levels of Red Guava. To conclude, it was found that the compound content was in the range 1.11-31.78 μg / ml for quercetin, and 0.87-31.84 μg / ml for ascorbic acid. The content of quercetin compounds is inversely related to the level of ripeness of the guava fruit. Conversely, the levels of ascorbic acid compounds increase with the maturity level of the guava fruit.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Author keywords” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Indexed keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Ascorbic acid,Guava,HPLC,Quercetin,Ripening[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Funding details” size=”size-sm” text_align=”text-left”][vc_column_text]This work was financially supported by Indonesia Endowment Fund for Education (LPDP), Indonesia. The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript. Many thanks to Integrated Laboratory Poltekkes Kemenkes Negeri Bandung, Indonesia.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”DOI” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][/vc_column_inner][vc_column_inner width=”1/4″][vc_column_text]Widget Plumx[/vc_column_text][/vc_column_inner][/vc_row_inner][/vc_column][/vc_row][vc_row][vc_column][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][/vc_column][/vc_row]