Article

Comparative Analysis of the Nutrient Composition of

Caulerpa lentillifera from Various Cultivation Sites
Wenchuan Zhou 1 , Yun Wang 2,3, * , Rui Xu 2 , Jialin Tian 2 , Ting Li 2 and Suwen Chen 2

1 Shenzhen Fisheries Development Research Center, Shenzhen 518067, China; [email protected]

2 Key Laboratory of Aquatic Product Processing, Ministry of Agriculture and Rural Affairs, South China Sea
Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China;
[email protected] (R.X.); [email protected] (J.T.); [email protected] (T.L.); [email protected] (S.C.)
3 Key Laboratory Efficient Utilization and Processing of Marine Fishery Resources of Hainan Province,
Hainan Engineering Research Center of Deep-Sea Aquaculture and Processing, Sanya Tropical Fisheries
Research Institute, Sanya 572018, China
* Correspondence: [email protected]; Tel.: +86-020-89221832

Abstract: The nutrient, amino acid, and fatty acid compositions of Caulerpa lentillifera from
various aquaculture regions were assessed to analyze their nutritional characteristics and
potential for aquaculture development. The nutrient composition of C. lentillifera was
determined according to the standard national nutrient determination methods of the
Association of Official Analytical Chemists, and the following data were revealed. (1) The
basic nutritional components of C. lentillifera were relatively more abundant in the three
aquaculture areas in Guangdong Province. The crude protein content in C. lentillifera was
measured at (8.70 ± 0.36)% and (18.57 ± 1.59)% for samples collected from the Dapeng and
Daya areas, respectively. These values were significantly higher compared to those obtained
from the Beihai, Dongshan, and Nha Trang regions (p < 0.05). Additionally, the crude
lipid content of C. lentillifera from the Daya population was measured at (5.10 ± 1.59)%,
which was significantly higher than that of samples from the Beihai, Dongshan, and
Nha Trang areas (p < 0.05). Furthermore, the total sugar contents in C. lentillifera were
quantified as (32.50 ± 4.22)% and (32.87 ± 2.59)% for samples from the Daya and Shanwei
regions, respectively. These values were significantly greater (p < 0.05) compared to
those observed in the other four populations. (2) The total amino acid (TAA) content
Academic Editor: Cornelia Witthöft of C. lentillifera ranged from 7.05% to 12.37%, with notable concentrations of the fresh
Received: 3 December 2024
amino acids aspartic acid (Asp) and glutamic acid (Glu). Significant variations in the
Revised: 20 January 2025 TAA and essential amino acid (EAA) levels were observed among the cultivation regions
Accepted: 29 January 2025 (p < 0.05), with the Shanwei population exhibiting the highest TAA content of 12.37% and
Published: 1 February 2025 EAA content of 4.65%, surpassing all other populations except for Guangdong Province
Citation: Zhou, W.; Wang, Y.; Xu, R.; (p < 0.05). (3) The fatty acid composition analysis revealed that the total fatty acid (TFA)
Tian, J.; Li, T.; Chen, S. Comparative and unsaturated fatty acid (UFA) levels in the long-stemmed grape fern alga from Daya
Analysis of the Nutrient Composition
were 2.400% and 1.048%, respectively, and significantly greater than those in the other
of Caulerpa lentillifera from Various
populations except for Dapeng (p < 0.05). These results imply that the nutritional quality
Cultivation Sites. Foods 2025, 14, 474.
https://doi.org/10.3390/
of the Daya population of C. lentillifera is relatively high. C. lentillifera exhibits a palatable
foods14030474 flavor profile, making it suitable for consumption and the development into high-quality
seafood condiments, thereby contributing to the environmentally sustainable advancement
Copyright: © 2025 by the authors.
Licensee MDPI, Basel, Switzerland. of C. lentillifera aquaculture.
This article is an open access article
distributed under the terms and Keywords: Caulerpa lentillifera; nutrient composition; amino acid; fatty acid
conditions of the Creative Commons
Attribution (CC BY) license
(https://creativecommons.org/
licenses/by/4.0/).

Foods 2025, 14, 474 https://doi.org/10.3390/foods14030474

Foods 2025, 14, 474 2 of 12

1. Introduction
Caulerpa lentillifera, commonly known as sea grape, is a type of green macroalgae that
falls under the categories of Chlorophyta, Ulvophyceae, Bryoposodales, Caulerpaceae, and
Caulerpa. C. lentillifera can be found throughout Southeast Asia, Okinawa Japan, Thailand,
Oceania, and South Korea, as well as in tropical and subtropical marine environments [1].
As reported in the literature, this green seaweed was initially recorded on the Red Sea coast
and later found in numerous other places, particularly in the Indo-Pacific region [2]. Its body
is bright green, and due to its solid apparatus, stolons, and upright branches, which are
distributed on symmetrical cysts, its shape resembles a bunch of grapes; thus, C. lentillifera
is also known as the sea grape [3]. In several Asian countries, C. lentillifera is often made
into a salad because of its tasty flavor. Moreover, C. lentillifera is rich in amino acids,
unsaturated fatty acids (UFAs), vitamins, minerals, and a variety of polysaccharides and it
has the advantages of low liquid and low calorie contents but high protein and dietary-fiber
contents. Additionally, C. lentillifera has a rich and juicy taste, similar to caviar; thus, it is
also known as green caviar [4,5]. Furthermore, C. lentillifera contains a variety of active
substances. Studies have shown that its extract has strong antioxidant activity and free
radical scavenging capacities; immunomodulatory and antitumor activities; anticoagulant
effects; and blood sugar and lipid regulatory properties [6–8]. The researchers identified
the monosaccharide components and structural characteristics of the polysaccharides
in C. lentillifera using infrared spectroscopy, high performance liquid chromatography
(HPLC), and nuclear magnetic resonance (NMR) analyses. Their findings indicated that
the polysaccharides predominantly comprised mannose, galactose, and xylose, with some
also containing glucose and glucosamine [9]. The critical active compound, caulerpin,
is a red bisindole alkaloid frequently found in various marine green algae, including
Caulerpa racemose, Caulerpa prolifera, and Caulerpa sertularioides [10]. Due to its rich nutrient
profile and bioactive compound composition, the demand for C. lentillifera has recently
increased in several Asian countries, leading to a rise in market prices. However, the
present production rate of C. lentillifera is insufficient to satisfy the existing demand. The
suboptimal cultivation conditions for the alga may contribute to its reduced productivity.
Consequently, it is imperative to determine the optimal culture conditions to enhance the
productivity of C. lentillifera.
In recent years, artificial breeding has been introduced in Shandong, Fujian, Guang-
dong, Hainan, Taiwan, and other regions via a variety of breeding modes, including sea
culture, land-based culture, and pond culture [11,12]. The growth of C. lentillifera has been
a major factor in the development of this species. To investigate the quality of C. lentillifera
cultured in different areas, the nutrient composition (moisture, crude protein, crude liquid,
ash, mannitol, and iodine), amino acid composition, fatty acid composition, and mass frac-
tions of C. lentillifera from six culture groups in four regions (Fujian, Guangdong, Guangxi,
and Vietnam) were determined, and the nutrient compositions were compared to better
understand the nutrients in C. lentillifera. This study aims to conduct a comprehensive eval-
uation of the nutritional value of C. lentillifera across various regions, thereby contributing
to the advancement of environmentally sustainable cultivation and utilization practices for
this species.

2. Materials and Methods

2.1. Sample Collection
C. lentillifera samples were collected from six farms in Guangdong, Fujian, Guangxi,
and Vietnam. The sampling locations are shown in Table 1. The samples were collected
between 17 June 2022 and 27 October 2022. They were transported to the laboratory in a
viable state, thoroughly rinsed with seawater to remove surface impurities, and prepared
Foods 2025, 14, 474 3 of 12

for experimental use. Subsequently, the samples were evenly divided into three portions,
weighed, and stored in a freezer at −80 ◦ C.

Table 1. Sample collection information.

Area Sampling Point Coordinates

Dapeng Bay of Penghu Sea area 114◦ 28′ 48′′ E, 22◦ 31′ 12′′ N
Guangdong Province Daya Bay of Huizhou 114◦ 31′ 12′′ E, 22◦ 34′ 48′′ N
Great Lake of Shanwei 115 34 1 E, 22◦ 49′ 42′′ N
◦ ′ ′′
Guangxi Province North Sea 109◦ 3′ E, 21◦ 25′ 48′′ N
Fujian Province Dongshan 117◦ 25′ 48′′ E, 23◦ 34′ 48′′ N
Vietnam Nha Trang 109◦ 19′ 12′′ E, 12◦ 10′ 48′′ N

2.2. Methods by Which the Basic Nutrient Composition Was Determined

The basic nutrient composition of C. lentillifera was determined according to the
standard methods of the Association of Official Analytical Chemists [13]. For moisture
determination, samples were dried at 105 ◦ C for 72 h to determine the moisture. The
dried samples were pulverized into a fine powder. Two grams of the powder were
utilized for ash measurement in a muffle furnace (FO610C, Yamato Scientific Co., Ltd.,
Tokyo, Japan) at 550 ◦ C, 1 g was employed for the determination of crude lipids us-
ing the Soxhlet extraction method (Soxtec 2055, FOSS, Höganäs, Sweden), and 0.2 g
were used for the assessment of crude protein via the Kjeldahl method (Kjeltec 8400,
FOSS, Hoganos, Sweden). Additionally, the following methods were used to evalu-
ate the other nutrients: total sugar content (GB/T 15672-2009) [14], crude fiber con-
tent (GB/T 5009.10-2003) [15], iodine content (GB 5009.267-2016) [16], mannitol content
(SC/T 3405-2018) [17], total amino acid (TAA) content (GB 5009.124-2016) [18], and fatty
acid content (GB 5009.168-2016) [19].
The TAA were quantified using the method of GB 5009.124-2016. The appropriate
amount of mixed samples (100 mg) was accurately weighed, and 5 mL of a 1:1 hydrochloric
acid solution was added to the hydrolysis tube. It was mixed thoroughly, and the tube was
placed in an electric blast thermostat set at 110 ± 1 ◦ C for hydrolysis over a period of 22 h.
After hydrolysis, the tube was removed and allowed to cool to room temperature. The
hydrolysis tube was opened, and the hydrolysate was filtered into a 10 mL volumetric flask.
The hydrolysis tube was rinsed several times with a small amount of water, transferring
the rinsing solution into the same volumetric flask. The volume was adjusted to the
calibration mark with water and mixed thoroughly. A pipette of 0.05 mL of the filtrate
was accurately measured into a 15 mL test tube and was evaporated to dryness under a
nitrogen stream. The residue was reconstituted in 2 mL of 0.02 mol L−1 hydrochloric acid
solution, with thorough mixing being ensured. The solution was passed through a 0.22 µm
microporous filter membrane before proceeding with the analysis. The quantification
of TAA was conducted utilizing an automated amino acid analyzer (LA8080, Hitachi,
Tokyo, Japan).
The fatty acid profile was performed using the method of GB 5009.168-2016. For fat
extraction, an appropriate quantity of the sample was weighed into a 100 mL cuvette,
followed by the addition of 8 mL of water. The mixture was thoroughly homogenized
before adding 10 mL of hydrochloric acid, ensuring thorough mixing. The flask was then
placed in a water bath maintained at 80 ◦ C for hydrolysis, which was conducted for 1 h.
The flask was agitated every 30 min to ensure that particles adhering to the flask walls
were incorporated into the solution. Upon the completion of hydrolysis, the sample was
removed and allowed to cool to room temperature. Subsequently, 10 mL of 95% ethanol
was added to the sample and mixed thoroughly. Fat extraction was performed using
Foods 2025, 14, 474 4 of 12

100 mL of ether in three separate batches, with the extracts being combined into a 100 mL
flat-bottomed flask. The ether layer was evaporated to yield the fat. For the saponification
of fat and the methyl esterification of fatty acids, to the fat extract, 4 mL of a 2% sodium
hydroxide methanol solution was added, and the mixture was incubated in a water bath
at 45 ◦ C for 30 min. Subsequently, 4 mL of a 14% boron trifluoride methanol solution was
added, and the mixture was again incubated in a water bath at 45 ◦ C for 30 min. After the
water bath treatment, the mixture was allowed to cool to room temperature. In a centrifuge
tube, 3 mL of n-hexane was added, and the mixture was subjected to extraction by shaking
for 2 min, followed by a period of settling to allow for phase separation. The supernatant
was collected and filtered through a 0.45 µm membrane for analysis. The samples were
analyzed using an Agilent 7890A gas chromatograph (Aglient Technologies, Palo Alto,
CA, USA).

2.3. Analysis of the Amino Acid Composition

The amino acid score (AAS) was calculated via the standard essential amino acid
(EAA) model score proposed by the FAO/WHO [20], and the amino acid model of egg
protein proposed by the Institute of Nutrition and Food Safety of the Chinese Academy of
Preventive Medical Sciences (IFSS) was used to calculate the chemical score (CS) [21] and
essential amino acid index (EAAI).

2.4. Data Analysis

There were three parallel treatment groups at each sampling point. The data are
expressed as the means ± standard deviations (x ± SDs). All the data were subjected
to one-way analysis of variance (ANOVA) in SPSS 26.0 to determine significance, and
differences between groups were analyzed by multiple comparisons via Duncan’s method,
and p < 0.05 was considered to indicate a significant difference.

3. Results
3.1. Comparison of the C. lentillifera Basic Nutrient Compositions
The basic nutrient compositions of C. lentillifera from different regions are shown in
Table 2. The C. lentillifera moisture contents were extremely high, close to 95%, and there
was no significant difference between the populations (p > 0.05). The ash determination
results revealed that the Nha Trang population had the highest ash content (65.50%),
the Shanwei population had the lowest content (22.83%), and the ash contents of the
Beihai, Dongshan, and Nha Trang C. lentillifera populations were significantly greater
than those of the three Guangdong populations (p < 0.05). In terms of crude protein
content, the Dapeng population had the highest (18.70%), the Nha Trang population had
the lowest (12.53%), and the populations followed the order Dapeng Bay > Daya Bay >
Shanwei > Dongshan > Beihai > Nha Trang; additionally, the crude protein contents of
the C. lentillifera samples from Dapeng Bay and Daya Bay were significantly greater than
the samples from the Beihai, Dongshan, and Nha Trang populations (p < 0.05). It was also
revealed that the Daya Bay population had the highest crude liquid content (5.10%), and
the Shanwei population had the lowest content (3.83%). The crude liquid content of the
Daya Bay population was significantly greater than that of the Beihai, Dongshan, and Nha
Trang populations (p < 0.05). Moreover, the Shanwei population presented the highest total
sugar content (32.87%), the Nha Trang population presented the lowest content (18.13%),
and the Daya Bay and Shanwei populations presented relatively high total sugar contents,
which were significantly greater than those of the other four populations (p < 0.05). The
mannitol determination results revealed that the DongShan population presented the
highest mannitol content (7.47%), the Dapeng Bay population presented the lowest content
Foods 2025, 14, 474 5 of 12

(5.30%), and the Shanwei and DongShan populations presented significantly greater
mannitol contents than the Dapeng Bay population (p < 0.05). Finally, the iodine content
of the Nha Trang C. lentillifera population was significantly greater than that of the other
populations (p < 0.05). Among the six C. lentillifera populations, the Guangdong population
presented a greater content of basic nutrients and was considered to be better quality than
the other three regional populations.

Table 2. Nutritional components of C. lentillifera cultivated in different areas (n = 3, dry weight, %).

Guangxi Fujian
Cultivation Guangdong Province Vietnam
Province Province
Area
Dapeng Bay Daya Bay Shanwei North Sea Dongshan Nha Trang
Moisture * 94.47 ± 0.51 94.83 ± 0.23 94.73 ± 0.81 94.87 ± 0.32 94.97 ± 0.35 94.57 ± 0.58
Ash 34.07 ± 1.34 b 25.80 ± 0.70 bc 22.83 ± 0.21 c 57.90 ± 8.84 a 59.60 ± 5.99 a 65.50 ± 7.11 a
Crude protein 18.70 ± 0.36 a 18.57 ± 1.59 a 17.20 ± 1.49 ab 13.57 ± 3.33 bc 14.17 ± 2.75 bc 12.53 ± 1.16 c
Crude liquid 4.57 ± 0.47 ab 5.10 ± 0.36 a 3.83 ± 0.81 b 4.67 ± 0.25 ab 3.93 ± 0.35 b 4.27 ± 0.40 ab
Total sugar 24.00 ± 3.38 b 32.50 ± 4.22 a 32.87 ± 2.59 a 19.93 ± 1.93 bc 20.00 ± 2.01 bc 18.13 ± 2.50 c
Mannitol 5.30 ± 1.15 b 6.17 ± 1.46 ab 7.17 ± 0.49 a 6.57 ± 0.40 ab 7.47 ± 0.25 a 6.07 ± 0.25 ab
Iodine 0.05 ± 0.02 ab 0.03 ± 0.01 c 0.04 ± 0.01 bc 0.04 ± 0.00 bc 0.05 ± 0.01 abc 0.07 ± 0.00 a
Notes: values with different letters indicate significant differences (p < 0.05); * indicates fresh weight.

3.2. Analysis of the Amino Acid Composition of C. lentillifera

The amino acid compositions and mass fractions of the different populations of
C. lentillifera are shown in Table 3. A total of 17 amino acids were detected in the
C. lentillifera samples; notably, tryptophan was not detected. The TAA content of
C. lentillifera ranged from 7.05% to 12.37%, the EAA content ranged from 2.89% to 4.65%,
the nonessential amino acid (NEAA) content ranged from 4.15% to 7.87%, and the flavor
amino acid (DAA) content ranged from 3.52 to 5.31%. The most abundant NEAA was
aspartic acid (Asp, ranging from 0.80 to 1.32%), followed by glutamic acid (Glu, ranging
from 0.85 to 1.21%). There were no significant differences (p > 0.05) among the TAA, EAA,
and NEAA contents of the three populations of long-stemmed grape ferns in Guangdong,
although the TAA and DAA contents in the Daya Bay and Shanwei populations were sig-
nificantly greater than those in the Beihai, Dongshan, and Nha Trang populations (p < 0.05).
Additionally, the EAA contents in the Dapeng Bay and Shanwei samples were significantly
greater than those in the Beihai, Dongshan, and Nha Trang samples (p < 0.05), while the
NEAA contents in the Dapeng Bay, Daya Bay, and Shanwei samples were significantly
greater than those in the Beihai, Dongshan, and Nha Trang samples (p < 0.05). Furthermore,
the EAA/TAA ratios of the six populations of long-stemmed grape ferns ranged from
35.87% to 41.03%, the EAA/NEAA ratio ranged from 55.93 to 69.63%, and the DAA/TAA
ratio ranged from 41.00–50.13%. The EAA/TAA, EAA/NEAA, and DAA/TAA ratios of
C. lentillifera from Beihai, Dongshan, and Nha Trang were approximately 40%, 70%, and
50%, respectively, with no significant differences among these three regions (p > 0.05), but
all were significantly greater than the corresponding ratios from the Guangdong population
(p < 0.05).
Foods 2025, 14, 474 6 of 12

Table 3. Amino acid composition of C. lentillifera cultivated in different areas (n = 3, dry weight, %).

Guangxi Fujian
Guangdong Province Vietnam
Amino Acid Province Province
Dapeng Bay Daya Bay Shanwei North Sea Dongshan Nha Trang
a a a b b
Thr * 0.77 ± 0.11 0.82 ± 0.11 0.81 ± 0.05 0.44 ± 0.07 0.40 ± 0.16 0.49 ± 0.11 b
Val * 0.57 ± 0.09 ab 0.57 ± 0.11 ab 0.68 ± 0.05 a 0.49 ± 0.07 ab 0.45 ± 0.15 b 0.54 ± 0.12 ab
Met 0.12 ± 0.06 0.15 ± 0.03 0.08 ± 0.04 0.12 ± 0.01 0.12 ± 0.04 0.12 ± 0.03
Ile * 0.47 ± 0.10 0.41 ± 0.08 0.51 ± 0.04 0.36 ± 0.06 0.35 ± 0.12 0.42 ± 0.10
Leu * 0.82 ± 0.20 0.78 ± 0.12 0.84 ± 0.06 0.62 ± 0.09 0.58 ± 0.20 0.70 ± 0.16
Phe *# 0.70 ± 0.12 ab 0.71 ± 0.09 ab 0.73 ± 0.04 a 0.48 ± 0.08 c 0.44 ± 0.16 c 0.52 ± 0.12 bc
Lys * 0.59 ± 0.07 ab 0.66 ± 0.10 a 0.63 ± 0.05 ab 0.49 ± 0.08 ab 0.43 ± 0.15 b 0.53 ± 0.14 ab
His * 0.30 ± 0.12 a 0.31 ± 0.03 a 0.35 ± 0.01 a 0.15 ± 0.03 b 0.12 ± 0.05 b 0.16 ± 0.05 b
Asp # 1.21 ± 0.15 ab 1.31 ± 0.19 a 1.32 ± 0.07 a 0.86 ± 0.13 c 0.80 ± 0.30 c 0.96 ± 0.18 bc
Ser 0.48 ± 0.07 ab 0.55 ± 0.07 a 0.51 ± 0.03 a 0.30 ± 0.05 c 0.29 ± 0.12 c 0.34 ± 0.09 bc
Glu # 1.10 ± 0.15 ab 1.18 ± 0.13 ab 1.21 ± 0.06 a 0.94 ± 0.16 ab 0.85 ± 0.28 b 1.05 ± 0.20 ab
Gly # 0.90 ± 0.19 abc 1.16 ± 0.19 a 1.07 ± 0.07 ab 0.81 ± 0.09 bc 0.67 ± 0.24 c 0.79 ± 0.14 bc
Ala # 0.29 ± 0.19 b 0.59 ± 0.11 a 0.60 ± 0.05 a 0.49 ± 0.07 ab 0.45 ± 0.15 ab 0.56 ± 0.12 a
Cys 0.26 ± 0.10 a 0.27 ± 0.04 a 0.23 ± 0.01 a 0.09 ± 0.01 b 0.08 ± 0.02 b 0.08 ± 0.01 b
Tyr # 0.37 ± 0.11 0.37 ± 0.05 0.30 ± 0.02 0.33 ± 0.05 0.31 ± 0.10 0.35 ± 0.09
Arg 0.46 ± 0.06 ab 0.58 ± 0.06 a 0.58 ± 0.03 a 0.44 ± 0.06 ab 0.38 ± 0.14 b 0.46 ± 0.09 ab
Pro 1.78 ± 0.21 a 1.86 ± 0.19 a 1.91 ± 0.14 a 0.38 ± 0.06 b 0.33 ± 0.11 b 0.40 ± 0.09 b
TAA 11.18 ± 1.68 ab 12.27 ± 1.66 a 12.37 ± 0.72 a 7.77 ± 1.15 c 7.05 ± 2.48 bc 8.44 ± 1.75 c
EAA 4.34 ± 0.84 ab 4.41 ± 0.65 ab 4.65 ± 0.32 a 3.15 ± 0.49 bc 2.89 ± 1.02 c 3.47 ± 0.80 abc
NEAA 6.84 ± 0.84 a 7.87 ± 1.01 a 7.73 ± 0.39 a 4.63 ± 0.67 b 4.15 ± 1.45 b 4.99 ± 0.98 b
DAA 4.57 ± 0.53 ab 5.31 ± 0.75 a 5.24 ± 0.25 a 3.90 ± 0.57 ab 3.52 ± 1.21 b 4.23 ± 0.83 ab
EAA/TAA 38.63 ± 1.63 b 35.87 ± 0.51 c 37.57 ± 0.45 b 40.50 ± 0.27 a 41.03 ± 0.21 a 40.87 ± 0.91 a
EAA/NEAA 63.00 ± 4.37 b 55.93 ± 1.29 c 60.10 ± 1.15 b 68.10 ± 0.72 a 69.63 ± 0.57 a 69.23 ± 2.61 a
DAA/TAA 41.00 ± 1.45 c 43.23 ± 0.42 b 42.30 ± 0.62 bc 50.13 ± 0.15 a 49.93 ± 0.29 a 50.10 ± 0.82 a
Notes: * Essential amino acids; # flavor amino acids. The values with different letters indicate significant
differences (p < 0.05).

3.3. A comprehensive Assessment of the Amino Acid Profile in C. lentillifera

The nutritional quality of the C. lentillifera cultivated in six different areas was evalu-
ated according to the EAA evaluation criteria proposed by the FAO/WHO, and the results
are shown in Table 4. The AASs revealed that the first C. lentillifera-limiting amino acid
was methionine (Met) + cystine (Cys), and the second limiting amino acid was isoleucine
(Ile); moreover, the amino acid with the highest score among the six different samples was
phenylalanine (Phe) + tyrosine (Tyr). Additionally, the chemical score data revealed that
the first limiting amino acid of C. lentillifera from all populations was Met + Cys, while the
second limiting amino acid varied among the six regions. The second limiting amino acid
of the Daya Bay and Beihai populations was Ile, which was in agreement with the AASs,
but the second limiting amino acid of the other four populations was lysine (Lys). The
amino acid with the highest chemical score differed among the six regions, as threonine
(Thr) gave the highest chemical score in the Guangdong region while Phe + Tyr gave the
highest scores in the other three region, which is consistent with the AASs. The highest
EAAI was found in the Nha Trang, Vietnam, population among the six regions, while the
lowest EAAI was found in the Dongshan, Fujian population.
Foods 2025, 14, 474 7 of 12

Table 4. Amino acid scores of C. lentillifera cultivated in different areas.

Fujian
Evaluation Amino Reference Guangdong Guangxi Vietnam
Province
Method Acid Proteins
Dapeng Bay Daya Bay Shanwei North Sea Dongshan Nha Trang
Thr 250 257.35 276.03 295.53 202.70 177.93 242.70
Val 310 190.51 192.99 248.29 225.74 197.07 269.28
Met + Cys 220 127.24 142.49 115.19 96.74 86.03 99.24
AAS Ile 250 155.98 139.13 185.32 167.37 152.96 207.80
Leu 440 272.96 262.57 306.43 287.15 257.34 347.42
Phe + Tyr 380 431.15 458.95 496.58 446.87 385.28 523.61
Lys 340 198.30 221.06 230.12 224.22 191.16 264.30
Thr 292 88.13 94.53 101.21 69.42 60.93 83.12
Val 411 46.35 46.96 60.41 54.92 47.95 65.52
Met + Cys 386 32.96 36.92 29.84 25.06 22.29 25.71
CS Ile 331 47.12 42.03 55.99 50.56 46.21 62.78
Leu 534 51.12 49.17 57.38 53.77 48.19 65.06
Phe + Tyr 565 76.31 81.23 87.89 79.09 68.19 92.67
Lys 441 44.96 50.13 52.18 50.84 43.35 59.93
EAAI 100 52.59 54.23 59.65 52.12 45.89 61.09

3.4. Analysis of the Fatty Acid Composition of C. lentillifera

The fatty acid compositions and mass fractions of different populations of C. lentillifera
are shown in Table 5. A total of 13 fatty acids were detected in C. lentillifera, including four
saturated fatty acids (SFAs), three monounsaturated fatty acids (MUFAs), and six polyun-
saturated fatty acids (PUFAs). The total fatty acid (TFA), SFA, MUFA, and PUFA contents
of the Daya Bay population were significantly different from those of the Shanwei, Beihai,
Nha Trang, and Dongshan populations (p < 0.05). Among the 13 fatty acids detected,
palmitic acid (ranging from 0.494 to 1.068%) and linoleic acid (0.264 to 0.392%) had the
highest contents, and the contents of these fatty acids were significantly greater in the Daya
population than in the other populations (p < 0.05). The ratios of UFAs to TFAs ranged
from 42.58 to 46.11%, with the highest ratios in the Beihai population and the lowest in the
Shanwei population.

Table 5. Comparative analysis of the fatty acid contents of C. lentillifera cultivated in different areas
(n = 3, dry weight, %).

Guangxi Fujian
Guangdong Province Vietnam
Province Province
Fatty Acid
Dapeng Bay Daya Bay Shanwei North Sea Dongshan Nha Trang
Myristic acids C14:0 0.022 ± 0.003 b 0.042 ± 0.004 a 0.036 ± 0.015 a 0.006 ± 0.002 c 0.009 ± 0.002 c 0.009 ± 0.004 c
Palmitic acids C16:0 0.886 ± 0.097 b 1.068 ± 0.029 a 0.853 ± 0.203 b 0.661 ± 0.055 c 0.537 ± 0.050 c 0.494 ± 0.041 c
Docosanoic acids C22:0 0.049 ± 0.005 a 0.050 ± 0.007 a 0.035 ± 0.006 b 0.028 ± 0.002 bc 0.020 ± 0.004 c 0.019 ± 0.004 c
Lignoceric acids C24:0 0.189 ± 0.019 a 0.192 ± 0.023 a 0.141 ± 0.026 b 0.117 ± 0.011 b 0.082 ± 0.013 c 0.082 ± 0.009 c
ΣSaturated fatty acids SFA 1.146 ± 0.123 ab 1.325 ± 0.055 a 1.065 ± 0.238 b 0.813 ± 0.069 c 0.648 ± 0.069 c 0.604 ± 0.050 c
Oleic acids (C18:1), n-9 * 0.069 ± 0.002 a 0.068 ± 0.004 a 0.071 ± 0.009 a 0.033 ± 0.004 b 0.025 ± 0.003 bc 0.023 ± 0.005 c
Erucic acids (C22:1), n-9 * 0.103 ± 0.014 a 0.096 ± 0.034 a 0.075 ± 0.010 ab 0.032 ± 0.009 c 0.021 ± 0.007 c 0.027 ± 0.012 c
ΣMonounsaturated fatty acids (MUFA) 0.079 ± 0.007 ab 0.084 ± 0.002 a 0.063 ± 0.020 bc 0.054 ± 0.006 cd 0.043 ± 0.005 d 0.051 ± 0.003 cd
Linoleic acids (C18:2), n-6 ** 0.264 ± 0.024 b 0.329 ± 0.011 a 0.256 ± 0.067 b 0.260 ± 0.037 b 0.194 ± 0.017 c 0.155 ± 0.014 c
Cis−8,11,14−eicosatrienoic acids (C20:3), n-6 ** 0.009 ± 0.001 b 0.013 ± 0.000 a 0.007 ± 0.002 c 0.006 ± 0.001 c 0.004 ± 0.001 d 0.005 ± 0.000 d
Arachidonic acids (C20:4), n-6 ** 0.075 ± 0.005 b 0.105 ± 0.002 a 0.067 ± 0.017 b 0.055 ± 0.007 bc 0.040 ± 0.003 c 0.038 ± 0.003 c
Cis−11,14,17−eicosatrienoic acids (C20:3), n-3 ** 0.007 ± 0.001 d 0.008 ± 0.000 cd 0.008 ± 0.001 cd 0.020 ± 0.003 a 0.012 ± 0.003 b 0.011 ± 0.002 bc
Eicosapentaenoic acids (C20:5; EPA), n-3 ** 0.055 ± 0.004 a 0.049 ± 0.001 a 0.037 ± 0.012 b 0.025 ± 0.003 c 0.017 ± 0.001 c 0.017 ± 0.001 c
Alpha linolenic acids (C18:3), n-3 ** 0.265 ± 0.029 ab 0.296 ± 0.007 a 0.206 ± 0.060 bc 0.210 ± 0.036 bc 0.153 ± 0.014 cd 0.133 ± 0.017 d
ΣPolyunsaturated fatty acids (PUFAs) 0.846 ± 0.057 ab 0.964 ± 0.053 a 0.727 ± 0.159 bc 0.641 ± 0.089 cd 0.466 ± 0.041 d 0.407 ± 0.037 d
TFA 2.072 ± 0.180 ab 2.400 ± 0.097 a 1.855 ± 0.413 bc 1.507 ± 0.160 cd 1.156 ± 0.115 de 1.062 ± 0.089 e
UFA/TFA 44.64% 43.66% 42.58% 46.11% 44.03% 43.13%
Notes: * monounsaturated fatty acids; ** polyunsaturated fatty acids; values with different letters indicate a
significant difference (p < 0.05).
Foods 2025, 14, 474 8 of 12

4. Discussion
4.1. Comparison of the C. lentillifera Basic Nutrient Compositions
The collection sites of the C. lentillifera samples in this study, which included the
regions of Guangdong, Guangxi, Fujian, and Vietnam, span a wide range of geographical
locations with environmental differences. In addition to moisture, the nutrient contents
of the different populations of C. lentillifera were significantly different (p < 0.05). The ash
content of the six populations of C. lentillifera ranged from 22.83 to 65.50%, which was
in agreement with the results of Syakilla [22], and the ash contents in Guangxi, Fujian,
and Vietnam were significantly greater than those in the three Guangdong populations,
indicating that C. lentillifera could enrich the inorganic ions in the water [23]. Moreover,
the crude protein, crude liquid, and total sugar contents of C. lentillifera ranged from
12.53 to 18.70%, 3.83 to 5.10%, and 18.13 to 32.87%, respectively, which was comparable
to the ranges of 12.5 to 14.76%, 0.78 to 2.32%, and 21.32 to 50.71% reported in the study
by Syakilla [22]. Furthermore, these values were essentially the same as those reported by
Tang et al. [24] (9.22% crude protein and 0.81% crude liquid) and Wang et al. [25] (14.9%
crude protein and 4.4% crude liquid). The crude protein, crude liquid, and total sugar
contents of the Guangdong population were greater than those of the Guangxi, Fujian, and
Vietnam populations, with the Daya Bay population being significantly different (p < 0.05)
from the Beihai, Dongshan, and Nha Trang populations. The growth of C. lentillifera is
affected by various factors, such as temperature, light, growth substrate, and growth cycle,
and the differences in nutrient contents may be related to the cultivation environment,
growth area, sampling time, and site [26]. Seaweeds, especially brown algae, are the main
raw material sources from which mannitol and iodine are extracted in China [27]. In this
study, the mannitol and iodine contents of C. lentillifera ranged from 5.30 to 7.47% and
0.03 to 0.07%, respectively, which was lower than those of kelp (12.3 to 15.1% and 0.36 to
0.47%, respectively) [28]. Thus, C. lentillifera is a suitable food to supplement the mannitol
and iodine required by the human body but it is not suitable for use as a raw material for
industrial extraction.

4.2. The Amino Acid Composition of C. lentillifera

Amino acids are the basic units of proteins, and the protein content and amino acid
composition and content of C. lentillifera should be considered when evaluating the nutri-
tional quality of this algae [29]. In this study, it was revealed that all six populations had
the same amino acid composition, which is consistent with other studies on the nutrient
composition of C. lentillifera [8]. When performing a nutritional evaluation, the FAO/WHO
ideal protein model recommends that the ratio of EAAs to TAAs should be approximately
40% and that the EAA/NEAA ratio should be greater than 60% [29]. The amino acid
compositions of five of the six regional populations, with the exception of the Daya popula-
tion, conformed to the ideal protein model, suggesting that C. lentillifera is a high-quality
protein source. Among the six regional populations in this study, the highest EAA/TAA
and EAA/NEAA ratios were found in the Dongshan regional population, indicating that
this population had highest quality protein, and the high contents of the DAAs Glu and
Asp in the six populations were the source of the distinctive seaweed flavor of C. lentillifera.
The highest DAA/TAA ratio was observed in the Beihai population, which indicated that
this C. lentillifera population had the best fresh flavor. The DAA/TAA ratio was the highest
in the Beihai population. Moreover, the proline (Pro) contents in the three Guangdong
populations were significantly higher than those in the Beihai, Dongshan, and Nha Trang
populations, and they differ from the amino acid contents reported in other studies. These
previous studies on the nutrient composition of C. lentillifera revealed that proline is an
osmotic regulator in plants and that environmental stresses cause an increase in the proline
Foods 2025, 14, 474 9 of 12

content in plants to improve plant resistance [30]. This study revealed that proline is an
osmoregulatory substance in plants. The high proline content of C. lentillifera in Guangdong
may be the result of an increase caused by environmental stress factors.
The level of satisfaction someone experiences from the food they consume is indicated
by the taste parameter [31]. Enhancing the aroma and taste of food can be achieved by
including glucose, sucrose, fiber, and other stimulants [32]. Non-volatile components in
food are primarily responsible for the taste part of flavor, for example, lipids, polysac-
charides, amino acids, and carbohydrates are responsible for the basic tastes of sweet,
sour, salty, bitter, and umami in seaweeds [33]. Seaweeds are known for their ‘umami’
taste, which is considered the fifth basic taste, and this flavor is marked by organic acid,
amino acid salts, and short peptides [34]. C. lentillifera from Daya Bay exhibited a higher
protein content compared to populations from other regions, characterized by elevated
levels of Asp (1.31 ± 0.19), Glu (1.18 ± 0.13), and gly (1.16 ± 0.19), which may enhance the
palatability of the seaweed samples. This suggests the potential for developing C. lentillifera
as a source of fresh flavorings.

4.3. A comprehensive Assessment of the Amino Acid Profile in C. lentillifera

The AAS results revealed that the first limiting amino acid of C. lentillifera was
Met + Cys, and the second limiting amino acid was Ile or Lys, which was essentially
consistent with the results of the study by Tang [24]. The EAAI is an indicator for evalu-
ating the nutrient composition of food, and a high EAAI indicates high nutritional value.
The EAAIs of the six populations in this study ranged from 45.89 to 61.09, among which
the Nha Trang population had the highest EAAI, and thus, the highest nutritional value.

4.4. The Fatty Acid Composition of C. lentillifera

C. lentillifera is low in liquid but rich in fatty acids, with a total of 13 fatty acids
detected, including six UFA species, with percentages ranging from 42.58% to 46.11%,
which are higher than those reported by Wang [8], who reported values of 37.09% and
36.74%, respectively. Among the six populations in this study, the Daya Bay population
presented the highest TFA and UFA contents, which were significantly greater than those
of the Shanwei, Beihai, Dongshan, and Nha Trang populations (p < 0.05), whereas the
Beihai population presented the highest percentage of UFAs. The fatty acid with the
highest content was palmitic acid, which is consistent with the composition of seaweeds
such as Gracilaria lichenoides and Meristotheca papulosa and the abundance of palmitic acid
in nature [35]. In addition to palmitic acid, C. lentillifera is rich in PUFAs, with high
contents of linoleic acid and Alpha linolenic acids, ranging from 0.155 to 0.329% and
0.133 to 0.296%, respectively. Linoleic acids and Alpha linolenic acids are essential fatty
acids that cannot be produced by the human body but are important for preventing
diseases and health maintenance [36]. In addition, the contents of arachidonic acids
and EPA were high, at 0.038–0.105% and 0.017–0.055%, respectively. Arachidonic acid is
essential for the development of the human brain and visual nerves and has important
physiological activities. Terrestrial plants and animals rarely contain EPA, as it is mainly
found in marine organisms, but EPA can reduce cholesterol and triglyceride contents and
can promote the metabolism of saturated fatty acids in the body, thus reducing blood
viscosity and preventing fat deposition in the blood vessel wall. It can also prevent
cardiovascular diseases such as atherosclerosis, cerebral thrombosis, hypertension, and
other cardiovascular diseases [37,38]. The World Health Organization’s guidelines on fatty
acids recommend that both adults and children decrease their intake of SFA to constitute
only 10% of their total energy consumption. Furthermore, it is advised to substitute
saturated fatty acids with PUFA in the diet, as this substitution positively influences the
Foods 2025, 14, 474 10 of 12

regulation of blood cholesterol and triacylglycerol levels, thereby significantly reducing

the risk of coronary heart disease [39,40]. According to the World Health Organization’s
healthy dietary recommendations, the desiccated material of C. lentillifera exhibits potential
for commercialization as a nutritious and appealing food product. For instance, snacks
derived from the dried substance of C. lentillifera could be developed.

5. Conclusions
This study utilized samples collected from six farms located in Guangdong, Fujian,
Guangxi, and Vietnam, and employed national standard methodologies for the quantifi-
cation of nutrient compositions, including moisture, ash, crude protein, crude lipid, total
sugar, mannitol, and iodine. The amino acid and fatty acid compositions of C. lentillifera
from various cultivation regions were analyzed. The results of this study revealed that
C. lentillifera is an edible seaweed rich in protein and total sugar but low in fat. It is a
natural and healthy food because it is rich in amino acids present in composition ratios in
accordance with FAO/WHO recommended standards. Additionally, C. lentillifera contains
a relatively high percentage of a variety of UFAs, in addition to iodine and mannitol. This
study demonstrated significant variation in the nutrient composition of C. lentillifera across
different cultivation regions. Notably, C. lentillifera sourced from the Daya Bay exhibited
superior nutritional value, suggesting enhanced health benefits. Additionally, C. lentil-
lifera possesses a favorable flavor profile, rendering it suitable for consumption and its
development into high-quality seafood condiment. This contributes to the environmentally
sustainable advancement of C. lentillifera aquaculture.

Author Contributions: W.Z.: designing and performing experiments; Y.W.: designing experiments,
writing and revising the manuscript; R.X.: collecting samples and statistical analyses; J.T.: review and
revising the manuscript; T.L.: collecting samples and statistical analyses; S.C.: collecting samples and
statistical analyses. All authors have read and agreed to the published version of the manuscript.

Funding: This research was funded the Hainan Provincial Natural Science Foundation of China
(323MS126), Guangdong Province Rural Science and Technology Commissioners Project, Guang-
dong Natural Science Foundation (2019A1515012159), Financial Special Project of the Ministry of
Agriculture and Rural Areas (NHYYSWZZZYKZX2020), Central Public-interest Scientific Institution
Basal Research Fund, CAFS (2020KX03, 2023TD97), Rural Revitalization Strategy Special Fund Seed
Industry Revitalization Project of Guangdong Province (2022SPY02001).

Institutional Review Board Statement: Not applicable.

Informed Consent Statement: Informed consent was obtained from all subjects involved in the study.

Data Availability Statement: The original contributions presented in the study are included in the
article, further inquiries can be directed to the corresponding author.

Acknowledgments: We thank the editor and anonymous reviewers for their comments that helped
improve this work.

Conflicts of Interest: The authors declare no conflicts of interest.

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