Special Issue: Novel Advances in Agricultural Science and Technology for Sustainable Farming in Tropical Region

Characterization of Amino Acids in Coffee Cherry Flour from Different Coffee Cultivation Areas: As Potential Functional Food

Roy Hendroko Setyobudi1, Shazma Anwar2, Mohammed Ali Wedyan3, Damat Damat1*,

Yogo Adhi Nugroho4, Tony Liwang4, Praptiningsih Gamawati Adinurani5, Satriyo Krido Wahono6,

Evika Sandi Savitri7, Bayu Agung Prahardika7, Irma Rahmaita Utarid8, Iswahyudi Iswahyudi9 and

Hemalia Agustin Rachmawati1

1University of Muhammadiyah Malang, Malang 65144, East Java, Indonesia; 2University of Agriculture, Peshawar 25130, Khyber Pakhtunkhwa, Pakistan; 3The Hashemite University, PO Box 330127, 13133 Zarqa, Jordan; 4Plant Production and Biotechnology Division, PT Smart Tbk., Bogor 16810, West Java, Indonesia; 5Merdeka University of Madiun, Madiun 63133, East Java, Indonesia; 6Research Center for Food Technology and Processing (PRTPP), National Research and Innovation Agency (BRIN) Special Region of Yogyakarta 55281, Indonesia; 7State Islamic University of Maulana Malik Ibrahim, Malang 65145, East Java, Indonesia; 8Central Research and Diagnostic “Satwa Sehat” Indonesia, Malang 65146, East Java, Indonesia; 9Universitas Islam Madura, Pamekasan 69317, East Java, Indonesia.

Received | May 12, 2024; Accepted | June 11, 2024; Published | July 05, 2024

*Correspondence | Damat Damat, Department of Food Science, Faculty of Agriculture and Animal Science, University of Muhammadiyah Malang, Jl. Raya Tlogo Mas No. 246, Malang 65144, East Java, Indonesia; Email: [email protected]

Citation | Setyobudi, R.H., S. Anwar, M.A. Wedyan, D. Damat, Y.A. Nugroho, T. Liwang, P.G. Adinurani, S.K. Wahono, E.S. Savitri, B.A. Prahardika, I.R. Utarid, I. Iswahyudi and H.A. Rachmawati. 2024. Characterization of amino acids in coffee cherry flour from different coffee cultivation areas: As potential functional food. Sarhad Journal of Agriculture, 39 (Special issue 1): 36-46.

DOI | https://dx.doi.org/10.17582/journal.sja/2023/39/s1.36.46

Keywords | Cascara, Circular economy, Coffee pulp, Environment friendly, Sustainable resource management, Waste to functional food

Copyright: 2024 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Coffee is one of the most widely consumed beverages worldwide, known for its stimulating effects and rich flavor. However, coffee processors generate a significant amount of waste material (Ballesteros et al., 2014; Corro et al., 2014; Das and Venkatachalapathy, 2016; Genanaw et al., 2021; Geremu et al., 2016; Li et al., 2023; Novita, 2016; Setyobudi et al., 2018, 2021, 2022), including solid waste, i.e., pulp (CP – from wet coffee method), or husk (CH – from dry coffee method). In recent years, efforts have been made to utilize this solid waste to create value-added products. CP and CH may employ coffee cherry tea or cascara (from Spanish cáscara, meaning husk or peel and pulp of a coffee cherry) with simple technology, especially for functional food. According to Ota (2018), cascara is a traditional beverage in Yemen (named Qishr), Ethiopia (Hashana), Bolivia (Sultana), and Costa Rica (Cáscara). Several researchers have also published on cascara studies (Ariva et al., 2020; Arpi et al., 2018; Heeger et al., 2017; Muzaifa et al., 2021; Nafisah and Widyaningsih, 2018; Novita et al., 2021; Pua et al., 2021; Rohaya et al., 2023; Zeckel et al., 2020). However, Setyobudi et al. (2019, 2021, 2022, 2023) concluded that it is more efficient and effective to recycle CP and CH to coffee cherry flour (CCF).

CCF is gaining attention as a potential functional food ingredient due to its high content of bioactive compounds, including polyphenols and dietary fiber (Corro et al., 2014; Damat et al., 2019; Reguengo et al., 2022; Setyobudi et al., 2019, 2022, 2023; Rohaya et al., 2023), which reported to possess various health benefits, such as antioxidant, anti-inflammatory and antimicrobial properties (Ballesteros et al., 2014; de Melo Pereira et al., 2020; de la Rosa et al., 2019; Dorsey and Jones, 2017; Geremu et al., 2016; Ifadah et al., 2021; Lestari et al., 2022) a attributed to the presence of phytochemicals, such as chlorogenic acids and melanoidins (Campos et al., 2021; Dorsey and Jones 2017; Janissen and Huynh, 2018).

Setyobudi et al. (2019, 2021, 2022) recommend using CCF as an iron (Fe) booster. Anemia is reported in high rankings, especially among adolescent girls and mothers in worldwide (Milman, 2011; Nurbadriyah, 2019) and in Indonesia (Bukhari et al., 2020; Ellie et al., 2012). Fe as a hemoglobin booster is also needed by people for auto-immunity and COVID-19 to increase their immune system (Das and Venkatachalapathy, 2016; Khadim and Al-Fartusie, 2023; Petek et al., 2020; Talebi et al., 2020). Several publications reported that the number of people with auto-immune diseases tends to grow globally (Illescas-Montes et al., 2019; Lerner et al., 2022; Miller, 2023). CCF will be of double benefit because it is a source of non-heme Fe, antioksidan, and gluten-free; a substance that auto-immune sufferers should avoid (Hansen et al., 2022; Illescas-Montes et al., 2019; Passali et al., 2020).

Several experts (Singh and Prasad, 2023; Shubham et al., 2020) stated that the absorption of non-heme Fe in the digestive tract of mammals is influenced by the presence of enhancing agents (also called boosters or promoters). One is an amino acid, which improves iron absorption by forming dissolved chelate (Fernández-Lázaro et al., 2020; Kanimozhi and Sukuma, 2023; Shubham et al., 2020). There are three types of amino acids, namely essential amino acids, semi-essential amino acids, and non-essential amino acids. Non-essential amino acids are amino acids that can be made in the body, also known as endogenous amino acids. In contrast, essential amino acids (indispensable amino acids) are amino acids that cannot be made in the body and can only be obtained by consuming foods that contain protein (Hubli et al., 2023; Winarno, 2008). There are 20 kinds of amino acids (Hubli et al., 2023; Trakalo et al., 2019) and several researchers (Bailey-Shaw et al., 2009; Bouafou et al., 2011; Penaloza et al., 1985; Tadese and Mebratu, 2017) stated that CP dry contains 17 various amino acids, with eight to nine are essential amino acids. Sera et al. (2000) stated that the amino acid composition in CP in the pulp was the same as in soybean (Glycine max (L.) Merr.) or Kapok (Ceiba pentandra (L.) Gaertn.) flour.

Setyobudi et al. (2021) compared amino acid levels of CCF Mengani, Bali, Indonesia to CCF-La Boite, a commercial product from Brazil. However, the manuscript reports the total amino acid levels, which needs to be more accurate because the important for health is essential amino acids, especially sulfuric amino acids, which act as boosters in the absorption of non-heme Fe (Andika et al., 2021; Trakalo et al., 2019; Kurniawati et al., 2022; Yuniastuti, 2014). Therefore, a comparative analysis of coffee flour from different sources is essential to understand the potential variations in its nutritional profile and health-promoting effects. This study aims to investigate the amino acid composition of coffee flour obtained from different coffee farms, including Ijen Farm at the side of Mount Ijen, Bondowoso, East Java; Karang Ploso Farm at the downhill Mount Arjuno, Malang, East Java; Mengani Farm at the slope of Mount Batur, Mengani, Bali and La Boitê, a commercial product created in Brazil - purchased from La Boite, a Manhattan business located at 724 11th Avenue, New York, NY 10019. Furthermore, this comparative analysis can contribute to the development of standardized production methods and quality control measures for CCF, ensuring consistency and reliability in its applications as a functional food ingredient.

Materials and Methods

Sample collection

Samples of Arabica coffee cherry were collected from three different sources: Ijen Farm (7°57’59.55”S, 114°01’14.37”E), Karang Ploso Farm (7°52’13.80”S, 112°34’54.44”E), Mengani Farm (8°17’16.63”S, 115°15’0.61”E), and La Boitê (40°45’59.6088”N, 73°59’37.5792”W), as CCF. Coffee cherries from three locations were dried to become cascara, then turned into powder with an electric coffee grinder K-500N (China). The powder is filtered by sieve size No. 80 mesh (local product). The filtered CCF is stored in a 7 cm × 10 cm CTIK plastic clip. Each sample was obtained in triplicate to ensure data reproducibility.

Sample preparation and analysis

Sample preparation: 1 g of CCF sample was weighed using an analytical balance laboratory- JA-3003 (China) and transferred it to a clean, dry container. The samples were properly labeled for identification.

Sample extraction: 10 mL of 0.1 M HCl (extraction solvent) was added to each container. The containers were sealed tightly and were vortexed for few minutes to ensure thorough mixing. The extraction was done according to method highlighted by Arpi et al. (2018).

Filtration and derivatization: The extracted sample solution was transferred to the filtration apparatus and filtered to remove any particulate matter or debris. The filtered sample solution was collected in a clean vial for further analysis. To derivatize, 10 μL of the sample and 10 μL of the internal standard were buffered to pH 8.8 in a total volume of 80 μL using borate buffer. The derivatization was initiated by addition of 20 μL reagent solution (6-amino-quinolyl-N-hydroxysuccinimidyl carbamate, AccQ. Flour, 3 mg mL–1 in acetonitrile). All chemical using pro analysis grade, Merck.

High-performance liquid chromatography (HPLC) analysis: The samples were heated to 55 °C for 10 min and then analyzed by HPLC (PerkinElmer® LC 300, USA) with fluorescence detection. The HPLC system consisted of a quaternary pump, a vacuum degasser, a thermostated auto sampler, a thermostated column department and a fluorescence detector. A small volume (20 μL) of the derivatized sample solution was injected into the HPLC column. The HPLC system was ran using the appropriate mobile phase and gradient conditions suitable for amino acid separation. The elution of amino acids was monitored using UV detector and recorded the chromatographic data.

Data analysis: The HPLC data was analyzed to determine the retention times and peak areas of individual amino acids. The amino acid concentrations were quantified by comparing the peak areas of the samples with standard calibration curves prepared using known concentrations of amino acid standards. The concentration of individual amino acids were calculated in the CCF samples based on the peak areas and calibration curves. Analysis of variance (ANOVA) was performed to assess the significance of differences in amino acid levels among the CCF samples (Adinurani, 2016).

Results and Discussion

Table 1 supports several previous researches (Bailey-Shaw et al., 2009; Bouafou et al., 2011; Penaloza et al., 1985; Tadese and Mebratu, 2017), there are 17 kinds of amino acids in CP which are processed into CCF. Aspartic acid, serine, glutamic, glycine, histidine, arginine, threonine, alanine, proline, cysteine, tyrosine, valine, methionine, lysine, isoleucine, leucine, and phenylalanine were among the amino acids identified. Eight categories of essential amino acids, two categories of semi-essential, and seven categories of non-essential. Two types of sulfuric amino acids (cysteine and methionine) which play an important role in the absorption of non-heme Fe (Andika et al., 2021; Trakalo et al., 2019; Kurniawati et al., 2022; Yuniastuti, 2014) were also found. A further study of ANOVA found the following findings:

 

Table 1: Concentrations of different amino acids in Coffee Cherry Fluor from different sources.

Parameters	Source of coffee cherry fluor - CCF				Anova test
	Ijen farm	Karang ploso farm	Mengani farm	La Boitê	F value	p value
Asp	0.61 ± 0.46	1.29 ± 0.01	0.56 ± 0.07	0.58 ± 0.01	2.36	0.213 ns
Ser	0.33 ± 0.00 b	0.26 ± 0.01 c	0.16 ± 0.00 d	0.80 ± 0.01 a	2589.53	0.000 **
Glu	1.56 ± 0.01 ab	1.83 ± 0.01 a	1.09 ± 0.13 c	1.37 ± 0.00 bc	22.78	0.006 **
Gly	1.54 ± 0.02 a	1.46 ± 0.01 a	0.29 ± 0.14 c	0.78 ± 0.01 b	68.01	0.001 **
His	0.03 ± 0.03 b	0.03 ± 0.03 b	0.02 ± 0.01 b	1.03 ± 0.01 a	385.67	0.000 **
Arg	0.52 ± 0.00 a	0.46 ± 0.01 ab	0.02 ± 0.01 b	0.03 ± 0.01 a	7.11	0.044 *
Thr	0.58 ± 0.00 b	0.65 ± 0.02 ab	0.27 ± 0.15 b	1.05 ± 0.02 a	17.55	0.009 **
Ala	0.63 ± 0.00 b	0.65 ± 0.02 ab	0.27 ± 0.15 b	1.05 ± 0.02 a	1.50	0.343 ns
Pro	0.57 ± 0.01 b	0.92 ± 0.00 a	0.31 ± 0.06 c	0.49 ± 0.00 b	72.28	0.001 **
Cys	0.01 ± 0.00 b	0.00 ± 0.00 b	0.00 ± 0.00 b	0.61 ± 0.04 a	283.82	0.000 **
Tyr	0.31 ± 0.01 a	0.31 ± 0.01 a	0.24 ± 0.05 a	0.09 ± 0.00 b	19.8	0.007 **
Val	1.18 ± 0.19 ab	1.38 ± 0.01 a	0.67 ± 0.09 bc	0.27 ± 0.01 c	21.40	0.006 **
Met	0.05 ± 0.03 b	0.08 ± 0.00 b	0.32 ± 0.11 b	0.96 ± 0.01 a	55.72	0.001 **
Lys	0.55 ± 0.00 a	0.64 ± 0.00 a	0.31 ± 0.06 b	0.26 ± 0.01 b	37.97	0.002 **
Ile	0.54 ± 0.00 ab	0.54 ± 0.01 ab	0.43± 0.08 b	0.72 ± 0.00 a	9.02	0.030 *
Leu	1.14 ± 0.05	1.59 ± 0.00	1.33 ± 0.04	0.75 ± 0.01	2.01	0.255 ns
Phe	0.54 ± 0.00 a	0.53 ± 0.00 a	0.25 ± 0.03 b	0.34 ± 0.01 b	90.05	0.000 **
Total AA	10.65 ± 0.25 a	12.70 ± 0.00 a	7.34 ± 0.89 b	10.73 ± 0.09 a	23.14	0.005 **

Asp, Aspartic acid; Ser, Serine; Glu, Glutamic; Gly, Glycine; His, Histidine; Arg, Arginine; Thr, Threonine; Ala, Alanine; Pro, Proline; Cys, Cysteine; Tyr, Tyrosine; Val, Valine; Met, Methionine; Lys, Lysine; Ile, Isoleucine; Leu, Leucine; Phe, Phenylalanine; Total AA, Total Amino Acids. * Significantly different (α = 5 %). ** significantly different (α = 1 %). Ns, not significantly different.

 

Table 1 provides data on various amino acids present in CCF obtained from different sources: Ijen Farm, Karang Ploso Farm, Mengani Farm and La Boitê. The analysis of the data showed that there is a significant 1 % in the 12 observed variables, a significant 5 % in the two observed variables, and a non-significant effect in the three observed variables.

Aspartic acid (Asp) did not show significant differences among the different sources of CCF. Non-essential amino acid aspartic acid is used in the production of proteins and the metabolism of other amino acids. The lack of variation in aspartic acid content among the sources could be attributed to the stability and uniformity in the processing methods used for CCF production across the farms (Clifford and Ramirez-Martinez, 1990).

Serine (Ser) content varied significantly, with the highest concentration found in La Boitê and the lowest in Mengani Farm. Serine is a metabolic amino acid that is conditionally necessary for a number of metabolic processes, including the generation of neurotransmitters and proteins (Dorsey and Jones, 2017). The observed variations in serine content could be attributed to factors such as genetic differences in coffee plant varieties, environmental conditions and post-harvest processing techniques used at different farms (Farah et al., 2006).

Glutamic (Glu) acid content differed significantly among the CCF with more concentration in Karang Ploso Farm and less in Mengani Farm. Glutamic acid is an important neurotransmitter and plays a crucial role in taste perception (Montavon et al., 2018). The differences in glutamic acid content may be influenced by coffee plant genetics, environmental conditions, and post-harvest processing, including drying methods (Pushpa et al., 2012), which may affected the amino acid composition of CCF.

Glycine (Gly) content showed significant differences among the sources, with maximum concentration in Ijen Farm and minimum in Mengani Farm. A non-essential amino acid called glycine is used in the production of proteins and a number of metabolic activities. The variations in glycine content may be affected by elements such as coffee plant genetics (Sunarharum et al., 2014), soil composition, and agricultural practices used at different farms (Montavon et al., 2018).

Histidine (His) content exhibited significant altrations among the sources, with the highest concentration in La Boitê and the lowest in Mengani Farm. Histidine is an essential amino acid that plays a crucial role in protein synthesis, tissue repair, and the production of histamine and carnosine (Ariva et al., 2020). Variations in histidine content can be attributed to coffee plant genetics, soil composition (Cheng et al., 2016) and agricultural practices.

Arginine (Arg) content differed significantly among the different sources of coffee flour, with more concentration in Ijen Farm and less in Mengani Farm. An amino acid known as arginine is a semi-essential component of several physiological functions, including immune system operation, hormone control, and cardiovascular health (Muzaifa et al., 2021). Variations in Arginine content may be influenced by factors such as coffee plant genetics, soil composition, and agricultural practices.

Threonine (Thr) content varied significantly, with the maximum concentration in La Boitê and the minimum in Mengani Farm. Threonine is an essential amino acid necessary for protein synthesis and the maintenance of healthy skin and connective tissues (Ciaramelli et al., 2019). Differences in threonine content may be due to variation in coffee plant genetics, environmental conditions, and post-harvest processing methods (Farah et al., 2006).

Alanine (Ala) did not show significant differences among the sources of coffee flour. A non-essential amino acid called alanine is important in the production of glucose and energy metabolism (Winarno, 2008). The similarity of alanine level in all sources could be attributed to the common metabolic pathways and stability of this amino acid during CCF production (Clifford and Ramirez-Martinez, 1990).

Proline (Pro) content exhibited significant variations among the sources, with more concentration in Karang Ploso Farm and less in Mengani Farm. Proline is a non-essential amino acid that is used to make proteins and aids in cellular defense and stress response (Ginz and Engelhardt, 2000). Variations in Proline content can be ascribed to coffee plant genetics, environmental conditions, and post-harvest processing techniques (Farah et al., 2006).

Cysteine (Cys) content altered significantly among the sources of coffee flour, with maximum concentration in La Boitê and minimum in Karang Ploso Farm and Mengani Farm. Cysteine is a semi-essential amino acid and a precursor for the antioxidant compound glutathione. Variations in cysteine content can be influenced by elements such as coffee plant genetics, environmental conditions, and processing methods, including drying (Ludwig et al., 2000).

Tyrosine (Tyr) content differed significantly among the sources, with the highest concentration observed in Ijen Farm and Karang Ploso Farm, while the lowest concentration was found in La Boitê. A non-essential amino acid called tyrosine is used to make hormones and neurotransmitters (Clifford and Knight, 2004). Variations in tyrosine content of coffee sources may be due to differences in coffee plant genetics, environmental conditions, and post-harvest processing methods.

Valine (Val) content showed significant variations among the sources of CCF, with more concentration in Karang Ploso Farm and less in La Boitê. Valine is an essential amino acid used in the synthesis of proteins and the production of energy (Machado et al., 2020). Variations in valine content can be influenced by factors such as coffee plant genetics, soil composition, and agricultural practices.

Methionine (Met) content varied significantly among different CCF sources with the maximum concentration in La Boitê and the minimum in Ijen Farm. Methionine is an essential amino acid involved in the production of proteins and other metabolic activities. Differences in methionine content may be affected by elements such as coffee plant genetics, environmental conditions (Romano et al., 2021) and post-harvest processing techniques.

Lysine (Lys) content differed significantly among the sources of coffee flour, with the highest concentration in Karang Ploso Farm and the lowest in La Boitê. Lysine is an essential amino acid that is crucial for protein synthesis, immune system health, and collagen formation (Liu et al., 2021). The causes of variations in Lysine content may be coffee plant genetics, environmental conditions and agricultural practices.

Isoleucine (Ile) content showed significant variations among the different sources of coffee flour, with maximum concentration in La Boitê and minimum in Mengani Farm. Isoleucine is an essential amino acid that is involved in the production of proteins and the control of energy. Variations in isoleucine content may be the result of variation in coffee plant genetics, soil composition and agricultural practices (Tadese and Mebratu, 2017). Coffee plant genetics play an important role, as coffee varieties possess unique genetic traits that affect amino acid biosynthesis pathways. Additionally, soil composition influences nutrient availability, impacting the plant’s ability to produce isoleucine. Agricultural practices, including fertilization techniques and pest management, can also influence the overall health and metabolic activity of coffee plants, consequently affecting isoleucine production (Machado et al., 2020).

Leucine (Leu) did not show significant differences among the different sources of CCF. Leucine is an essential amino acid that is involved in protein synthesis and is important for both muscle development and repair. The similar content of Leucine among the sources could be attributed to the common metabolic pathways and stability of this amino acid during CCF production (Clifford and Ramirez-Martinez, 1990). The stability of leucine during coffee production processes may contribute to its uniformity. Leucine resistance to degradation during roasting and other processing steps could result in its consistent levels (Rohaya et al., 2023). Furthermore, leucine shares metabolic pathways with other amino acids that might experience fluctuations due to environmental and processing factors. While coffee varieties and growth conditions influence amino acid compositions, leucine’s essential role in biological processes might necessitate a more regulated synthesis, maintaining its levels across diverse sources (Jo et al., 2023).

Phenylalanine (Phe) content differed significantly among the sources of coffee flour, with the highest concentration in Ijen Farm and the lowest in Mengani Farm. Phenylalanine is an essential amino acid and a precursor for various neurotransmitters and hormones (Clifford and Knight, 2004). Variations in Phenylalanine content may be due to coffee plant genetics, environmental conditions and post-harvest processing techniques. The genetic variation can influence the enzymatic pathways responsible for phenylalanine biosynthesis (Cheng et al., 2016). Genetic traits can affect the expression of enzymes involved in the shikimate pathway, which is a metabolic pathway leading to the synthesis of phenylalanine and other aromatic compounds.

Total Amino Acids (Total AA) content varied significantly among the sources of coffee flour, with more concentration in Karang Ploso Farm and less in Mengani Farm. The total amino acid content reflects the overall composition of amino acids in CCF and is affected by a combination of factors such as coffee plant genetics, environmental conditions, agricultural practices, and post-harvest processing methods (Reguengo et al., 2022). Genetic variations can lead to differences in the abundance and composition of amino acids in coffee beans (Ciaramelli et al., 2019). The significant variation in the amino acid composition could also be attributed to the nutrient availability in the soil that directly affects amino acid synthesis as proper fertilization practices can provide the necessary nutrients for optimal growth and amino acid production. Girma et al. (2020) reported that the biochemical composition of the coffee varies with coffee varieties and growing altitudes. Furthermore, different coffee varieties have distinct genetic makeup, influencing their response to microclimate (Murkovic and Derle, 2006). Some varieties might have genetic traits that lead to higher biochemical production, better adaptation to altitude-related stress, or different pathways for producing secondary metabolites.

Conclusions and Recommendations

The research work concludes that CCF from La Boite performed better as compared to CCF from other three locations of Indonesian (Ijen Farm, Karang Ploso Farm, and Mengani Farm). This finding must be followed up by studying why CCF from La Boite farm outperforms the others. Indonesia, being the fourth-largest coffee-exporting country in the world, primarily relies on smallholder coffee production. By learning the correct CCF preparation method will (i) minimize environmental pollution, (ii) provide gluten-free CCF with a low glycemic index and as a source of non-heme iron, (iii) reduces export of wheat flour (iv) boost the income of farmers as well as coffee stakeholders and (v) stimulate economic circulation.

Acknowledgments

With sadness, the authors report Yogo Adhi Nugroho, one of the authors of this article, passed away after a fight against COVID-19. We sincerely appreciate his enthusiasm and dedication to the writing of this manuscript, especially the statistical analysis. The authors are grateful to Ricky Hendarto Setyobudi and Fitri Ramli, Arabica Coffee Factory at Mengani, Kintamani, Bali, Soni Sisbudi Harsono, University of Jember, and Pandu Prabowo, Kopi Carlos, Malang for supplying the CCF as research material. Also, thank you to Yohannes Kohar Whisnu Wardhana, which has brought to Indonesia six CCF packages from La Boite, a store at Manhattan, 724 11th Avenue New York, NY 10019. The authors thank the Directorate of Research and Community Service, University of Muhammadiyah Malang (DPPM – UMM), which funded this research with letter No. E.2a/811/BAA-UMM/viii/2023.

Novelty Statement

Prior studies (Bailey-Shaw et al., 2009; Bouafou et al., 2011; Penaloza et al., 1985; Tadese and Mebratu, 2017) have documented the presence of amino acids in coffee pulp (CP). However, these earlier investigations focused solely on CP from a single coffee cultivation area. This current research shows novelty because it compares amino acids in CP coffee cherry fluor (CCF) from three distinct coffee cultivation areas of Indonesia with commercially available CCF from Brazil. Furthermore, this research establishes a link between the presence of amino acids and their potential role in boosting hemoglobin levels to combat anemia.

Author’s Contribution

Roy Hendroko Setyobudi: Conceptualized and designed the study, elaborated the intellectual content, performed literature search, manuscript preparation, and manuscript revision.

Shazma Anwar: Performed literature search, manuscript preparation, and manuscript revision.

Damat Damat: Research supervision, and elaborated the intellectual content.

Tonny Liwang and Irma Rahmaita Utarid: Amino acid analysis.

Yogo Adhi Nugroho and Praptingsih Gamawati Adinurani: Statistical analysis.

Satriyo Krido Wahono, Mohammed Ali Wedyan, Evika Sandi Savitri, and Bayu Agung Prahardika: Performed the literature search and manuscript review.

Iswahyudi Iswahyudi and Hemalia Agustin Rachmawati: Administration, Turnitin check, Grammar check, rewrite.

Conflict of interest

The authors have declared no conflict of interest.

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