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Proximate analysis of mulberry fruits

The statistical differences in terms of moisture, ash, pH, TAC, TSS, TDW, crude protein, and crude fat contents of each mulberry cultivar were showed in Table 1. The moisture considered in this study varied from 70.0% (Yaan 3) to 87.4% (Guang ) and the fruits of Morus atropurpurea Roxb. tend to have comparable higher moisture contents for Zhongsang , Dashi, Guang , and Jushensang cultivars were all above 85%. The moisture of fresh mulberry grown in the East Anatolia Region of Turkey ranged from 71 to 75% with a very low variation among the three different cultivars (Morus alba, Morus rubra, and Morus nigra).[1]

Table 1.

Proximate composition of different mulberry cultivars*

Ash values were significantly different amongst the cultivars and lowest contents were found for Morus multicaulis Perr. with the values of 3.64% (Damo4××guiV-4) and 3.48% (Hongyayizhilaishisheng99-6), respectively. Compared to other cultivars, Morus atropurpurea Roxb. had higher ash content for Zhongsang , Dashi, Guang , and Jushensang cultivars with 5.73%, 5.24%, 6.28%, and 6.75%, respectively.

The pH of the eight mulberry fruits were from 3.37 (Jushensang) to 5.33 (Lvshenzi) and the Morus multicaulis Perr. has relatively high pH values of 4.80 (Hongyayizhilaishisheng99-6) and 4.44 (Damo4××guiV-4). These values were consistent with previously reported results for mulberry cultivars grown in Turkey (pH 3.52-5.60).[1,16] As for TAC, the value was from 0.11% (Lvshenzi) to 1.82% (Jushensang). Both the amounts and compositions of the organic acids were found to be variable for the tested species, the main organic acids were citric and malic acids, a trace amounts of tartaric and ascorbic acid.[2] It might be presumed that the different contents of these organic acids in different mulberry cultivars might be related with the variation of their pH.

As for TDW, the cultivar of Yaan 3 showed the highest value 30.0%, followed by Lvshenzi, Hongyayizhilaishisheng99-6, and Damo4××guiV-4 with 21.1%, 19.4% and 17.4%, respectively. It could be observed that the cultivars of Morus atropurpurea Roxb. has comparable low TDW compared with other cultivars which might be related with their high moisture contents. The TSS values were significantly different amongst the cultivars varied from 12.5% (Guang ) to 22.4% (Lvshenzi). The cultivars of Morus atropurpurea Roxb. had relatively low TSS at average 13.0% except Dashi (18.7%).

The variation of TDW, TSS, pH, and TAC in different mulberry cultivars could be result of heterozygote nature of seed propagated genotypes, different species, the climate, growth and geographical conditions where grown.[17,18] According to the results, the Lvshenzi cultivar (Morus alba L.) fruit might be recommended for processing due to relatively high TSS and TDW contents. High fruit TSS and acid content is a combination for fruit juice process found only in a few other fruits such as pomegranate and kiwifruit. Therefore, the Dashi cultivar seems the most promising, combined for more bright and red fruits appreciably high TSS and acid content, which may be especially useful in developing cultivars with greater agronomic potential. The Morus atropurpurea Roxb. might be proposed for fresh fruit production, since it has attractive full fruits with flesh plump, bright color, bigger particle, and higher moisture.

The crude protein contents of the eight mulberry cultivars were given in Table 1. The cultivar Yaan 3 was found to be the highest protein values (5.54%) followed by Dashi (4.71%), Lvshenzi (2.99%), Damo4××guiV-4 (2.41%), Guang (2.30%), Hongyayizhilaishisheng99-6 (2.24%), Jushensang (1.87%), and Zhongsang (1.62%). The crude protein content of Yaan 3 was appreciably higher than other mulberry cultivars and thus was interesting for food products and dietary consumption.

The crude fat contents of the mulberry cultivars were low and ranged from 0.23% (Damo4××guiV-4) to 2.33% (Lvshenzi) as shown in Table 1. As for the Morus atropurpurea Roxb., the cultivars of Zhongsang and Jushensang have relatively high crude fat content 1.16 % and 1.05%, respectively. The results were close to a previous study that reported the total fat contents of the mulberry species were low, between 0.85% (Morus. rubra) and 1.10% (Morus. alba).[1]

The fatty acid composition

The fatty acid composition analysis was performed by GC-MS for the oil samples from mulberry fruits. There were 16 fatty acids found in test cultivars and significant variation showed in Table 2. Linoleic acid (C18:2) was the dominant fatty acid in all mulberry cultivars, ranged from 26.40% to 74.77%, followed by palmitic acid (C16:0) with the value 9.29%-22.26%. Linoleic acid is one of the two essential fatty acids that humans require because they cannot be produced by the human body. Deficiency symptoms may include dry hair, hair loss, and poor wound healing.[19]

Table 2.

Fatty acid composition of different mulberry cultivarsa

All cultivars contained stearic acid (C18:0) with the values of 3.78%-10.48%. Oleic acid (C18:1) was found with relatively high amount in Dashi, Guang , Damo4××guiV-4, and Hongyayizhilaishisheng99-6, whereas not detected in other cultivars. Our results were comparable to the finding reported by Ercisli and Orhan,[1] who reported that the fatty acids found at the highest levels in three black mulberry fruits were linoleic acid (43.4%-61.9%), palmitic acid (12.1%-24.8%), and oleic acid (2.33%-16.01%), respectively.

Myristic acid (C14:0) was detected as minor amounts in Zhongsang , Dashi, Hongyayizhilaishisheng99-6, and Yaan 3 fruits with 0.27%, 0.58%, 0.49%, and 0.18%, respectively. Palmitoleic acid (C16:1), margaric acid (C17:0), and linolenic acid were only found in Zhongsang , Hongyayizhilaishisheng99-6, and Yaan 3 fruits. Behenic (C22:0) acid (0.20%-3.04%) and tetracosanoic (C24:0) acid (0.78%-3.39%) were detected in all mulberry cultivars except Lvshenzi and eicosanoic acid (0.53%-2.45%) was not found in Guang . 11-Eicosenoic acid (C20:1) was only found in Zhongsang (0.49%) and Yaan 3 (0.84%). Hongyayizhilaishisheng99-6 was the only cultivar that contained brassidic acid (C22:1), azeloic acid (C11:0), and oxiraneoctanoic acid (C19:0) with the value of 0.23%, 0.41%, and 0.83%, respectively.

Linoleic acid, palmitic acid, and oleic acid generally constitute the major fatty acids of fruits. In any case, the fatty acid composition of the mulberry cultivars was of some similar (major compounds including palmitic, linoleic, stearic, behenic acid, tetracosanoic acid, and eicosanoic acid) in the mulberry cultivars studied and there were also appreciable differences among the fatty acids of minor amount. The differences among fatty acids might mostly depend on the species and physiological environments.

Mineral elements

The knowledge of the levels of trace elements in mulberry fruits is necessary because of their benefits for human health. Mulberry was a good source of minerals which could provide nutritionally useful amounts of most of them including K, Ca, Na, Mg, Cu, Fe, P, Mn, and Zn [Table 3]. There were significant differences in the levels of minerals among the test mulberry cultivars. The content of K was especially higher than other elements from 521.37 mg/100g DW (Hongyayizhilaishisheng99-6) to .60 mg/100g DW (Guang ). The Na content was low and the K/Na ratio was relatively high which has been considered to be an advantage from the nutritional point of view, since the high content of K can be utilized beneficially in the diets of people who take diuretics to control hypertension and suffer from excessive excretion of K.[20]

Table 3.

Minerals contents of different mulberry cultivars*

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The content of P in mulberry fruits was also remarkable with the range of 133.97-294.95 mg/100g DW. The cultivar Jushensang showed the highest level (294.95 mg/100g DW) and the value was lower than that reported in strawberry.[21] Ca is important for bone growth and neurologic function, whereas Mg is required by many enzymes as cofactors and its dietary deficiency is linked with ischemic heart disease.[22] The relatively high amounts of Ca and Mg were also found in these fruits, and especially the Yaan3 cultivar, showed the highest contents, 296.24 mg/100g DW and 202.22 mg/100g DW, respectively.

Fe, Cu, Zn, and Mn are other minerals of nutritional importance in mulberry. Higher variations were observed for Fe among the mulberry cultivars ranged from 2.490 mg/100g DW (Damo4××guiV-4) to 6.500 mg/100g DW (Yaan 3). The Cu content varied from 0.162 mg/100g DW (Hongyayizhilaishisheng99-6) to 0.680 mg/100g DW (Yaan 3) with some exceptions for Zhongsang , Guang , and Damo4××guiV-4. Zn is especially important for the normal functioning of the immune system, whereas Mn could activate several enzymes which protect cells from free radicals attack and regulation of glucose homeostasis. The Zn and Mn contents among different cultivars were in the range of 1.108-4.066 mg/100g DW and 0.614-2.608 mg/100g DW, respectively. Their levels were very similar to other fruits such as strawberry, blackcurrant, redcurrant, and raspberry, etc.[23] Meanwhile, the Se, As, or Cr contents of all mulberry fruits were very low and the Se content was much higher compared to As. No Cr was detected in the five mulberry fruits such as Zhongsang , Dashi, Guang , Damo4××guiV-4, and Lvshenzi.

The mineral composition of mulberry fruits not only depended on the species or cultivars, but also on the growing conditions such as soil, climatic, geographical conditions, the addition of fertilizers, and cultural management techniques.[24] The results indicated that mulberry was a better source of minerals and could be recognized as a valuable horticultural product based on their rich and beneficial nutrient composition.

Total anthocyanins, polyphenols, flavonoids, and sugars content of mulberry cultivars

The total anthocyanins, polyphenols, flavonoids, and sugars content among different mulberry cultivars were statistically significant as given in Table 4. The total anthocyanins contents were from 19.00 mg/100g (Yaan 3) to 193.00 mg/100g (Jushensang), which were not detected in Lvshenzi. The Morus atropurpurea Roxb. was observed to be higher in anthocyanins content compared to other cultivars. Dashi contained comparable high amounts of anthocyanins (155.00 mg/100 g) which was in agreement with that reported by Song et al,[25] who also presented that the contents of anthocyanins in different species varied widely, and not only the cultivars, but also the plant growing environment could affect anthocyanins content.

Table 4.

Contents of total anthocyanins, polyphenols, flavonoids and sugars of mulberry cultivars*

Mulberry was also a rich source of polyphenols and the total polyphenols clearly showed that fruits of Morus atropurpurea Roxb. (189.67-246.00 GAE mg/100mg DW) had high total polyphenols content and their wide differences among cultivars. Bae and Suh[26] found the similar amounts of total polyphenols in mulberries cultivated in Korea. The variation of phenolic compounds in the fruits depends on many factors such as the degree of maturity at harvest, genetic differences, and environmental conditions, etc. In red-colored fruits, phenolics increase during the last ripening stage due to the maximal accumulation of anthocyanins and flavonols.

Total flavonoids content in different cultivars widely varied from 40.94 RE mg/100g DW (Yaan 3) to 150.32 RE mg/100g DW (Hongyayizhilaishisheng99-6) and the exception was Lvshenzi cultivar. Previously published results of total flavonoids content of mulberry fruits in Turkey was higher than the results presented here probably because of the different species, growth areas, and extraction methods.[26]

Total sugars content is very important from a sensory viewpoint because it confers sweetness to the fruit and sweet mulberry are particularly appreciated for this sensory attribute. Among the considered cultivars, the Yaan 3 showed a total sugars content higher than the others. There was no obviously difference in Morus multicaulis Perr., and the total sugars contents of Damo4××guiV-4 and Hongyayizhilaishisheng99-6 were .24 DE mg/100g DW and .35 DE mg/100g DW, respectively. The Morus atropurpurea Roxb. was observed to be moderate level of total sugars (at average .21 DE mg/100g) and the lowest content was detected in Guang with value of .72 DE mg/100g [Table 4]. It could be observed that mulberry fruit contains moderate amounts of sugars compared to sweet cherry which averagely contains 150-230 g/kg.[27] It was slightly higher than that presented in previous study which showed the average sugar contents of M. nigra and M. rubra were 12.2% and 7.9%, respectively.[2]

Antioxidant activity

Phenolic compounds including anthocyanins, flavonoids, and phenolic acids are known to be responsible for antioxidant activities in fruits and the fruits with higher phenolic contents generally show stronger antioxidant activities.[28,29] The concept of antioxidant activities which describes the ability of different food antioxidants in scavenging preformed free radicals has been suggested as a tool for investigating the health effects of antioxidant-rich foods. Based on the DPPH and OH radicals scavenging assays, the mulberry extracts obtained from the eight cultivars had considerable high antioxidant activities [Figure 1]. The RSA of DPPH among the mulberry cultivars varied from 50% to 96%. The comparable high RSA was found in the cultivars Jushensang (96%) and Zhongsang (93%) in Morus atropurpurea Roxb., while Hongyayizhilaishisheng99-6 also had high DPPH RSA of 88%. The mulberry cultivars were found to have better ability to scavenge hydroxyl radicals and the four cultivars of Morus atropurpurea Roxb. exhibited considerable high RSA (92%-97%) with the same result of the DPPH assay. The result indicated that Jushensang exhibited the highest antioxidant activity, while the lowest was found in Lvshenzi which showed the lowest RSA values for both DPPH and OH radical scavenging assays (50.5% and 68.9%, respectively). It has been reported that the phytochemicals responsible for antioxidant activity most likely be accounted for phenolics (including non-anthocyanin phenolics), anthocyanins, and other flavonoid compounds.[30] Therefore, the antioxidant activity observed in the Lvshenzi cultivar which gave the absence of anthocyanins and flavonoids could be due to the small amount of total polyphenols content.

Figure 1.

The scavenging radical ability of different mulberry cultivars on DPPH and OH. *1-Zhongsang , 2-Dashi, 3-Guang , 4-Jushensang, 5- Damo4××guiV-4, 6- Hongyayizhilaishisheng99-6, 7-Yaan 3, 8-Lvshenzi

It had been reported relatively higher levels of antioxidant activity in highly pigmented berries, pomegranate juices, red wine, green tea infusion,[31&#;33] and blueberries constituted one of the richest reported antioxidants.[34] Our results indicated that mulberry fruits could also be considered as a potential resource for antioxidants from dietary fruits consumption and the development of functional drinks.

Correlations between antioxidant activity and the contents of total anthocyanins (TA), total polyphenols (TP), total flavonoids (TF) and total sugars (TS)

Correlations between the antioxidant activities and the contents of TA, TP, TF, and TS were shown in Table 5. There was a good linear correlation between the antioxidant activity and TP in the extracts of mulberry determined by DPPH and OH radical scavenging assays. Previous literatures had shown that anthocyanins (especially monoglucosides of cyanidin and delphinidin) as well as non-anthocyanin phenolic (chlorogenic acid, kaempferol, quercetin, etc.) possess high antioxidant activity which could compare well with the activity of the well-known antioxidants α-tocopherol and trolox.[35] The antioxidant activity values were slightly better correlated to total polyphenols (TP vs. DPPH, R2= 0.: TP vs. OH, R2 = 0.) rather than to total anthocyanins (TP vs. DPPH, R2= 0.: TP vs. OH, R2= 0.). This conclusion was in agreement with previous findings obtained on blueberries and on red wines.[33,34] The correlations between the antioxidant activity and total flavonoids were TF vs. DPPH, R2=0. and TF vs. OH, R2=0., respectively. There was a negative linear correlation coefficient between the antioxidant activity and total sugars (TS vs. DPPH, R2=0.; TS vs. OH, R2=0.). There was a slightly better correlation between antioxidant activity and TP/TA as compared to that of TF/TS, which meant that polyphenols and anthocyanins significantly contributed to the antioxidant activities of mulberry fruits.

Table 5.

Correlations between antioxidant activity and the total anthocyanins, polyphenols, flavonoids and sugars

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