HPLC-MS/MS法测定甜樱桃花色苷与非花色苷酚的组成与含量

应用高效液相色谱-质谱联用技术测定甜樱桃‘雷尼’、‘红艳’、‘红灯’3个品种的花色苷与非花色苷酚的组成与含量。花色苷的检测条件为:色谱柱Kromasil 100-5C18柱(250 mm×4.6 mm,6.5μm),流动相为水-甲酸-乙腈溶液,梯度洗脱,进样量30μL,流速1.00 m L/min,柱温50℃,检测波长525 nm;非花色苷酚的检测条件为:色谱柱Zorbax SB-C18(50 mm×3.0 mm,1.8μm),流动相为1%乙酸-1%乙酸-乙腈溶液,梯度洗脱,进样量2μL,流速1.00 m L/min,柱温25℃,检测波长280 nm。结果表明,3个品种共检测到9种花色苷,主要为花青素-3-芸香糖苷和花青素-3-葡萄糖苷,其在‘红艳’果皮、‘雷尼’果皮、‘红灯’果皮、‘红灯’果肉中的含量分别为5.21、2.51、75.70、7.40 mg/g和0.09、0.07、3.57、0.34 mg/g。非花色苷酚类化合物检测出了芦丁与山柰酚-3-芸香糖苷这2种化合物,其在‘红艳’果皮、‘雷尼’果皮、‘红灯’果皮中的含量分别为0.30、0.63、0.74 mg/g和1.17、2.91、2.37 mg/g。     

Differentiation of raspberry varieties according to anthocyanin composition

 Four Spanish raspberry (Rubus idaeus L.) cultivars, Autumn Bliss, Heritage, Rubi, and Ceva, were studied in order to determine, qualitatively and quantitatively, both anthocyanins and vitamin C composition, owing to their significance as dietary compounds with antioxidant activity and also to the relation with color quality. Raspberry anthocyanin composition, determined by HPLC and GC-MS, was characteristic to each cultivar. The higher anthocyanin content expressed as cyanidin-3-glucoside was found in the late cultivars, Rubi (96.08 mg/100 g f.w.) and Ceva (122.88 mg/100 g f.w.). Three cultivars showed cyanidin-3-sophoroside (63.86–21.91 mg/100 g f.w.) and cyanidin-3-glucoside (25.12–14.00 mg/100 g f.w.) as the major pigments, while Autumn Bliss had a similar relative amount of cyanidin derivatives, cyanidin-3-rutinoside being the pigment in major concentration (10.53 mg/100 g f.w.). The vitamin C contents were determined by HPLC. The Rubi cultivar showed the highest amount of vitamin C (31.14 mg/100 g f.w.). The organic acids were determined by HPLC as fruit constituents related to color quality. Citric acid was the main non-volatile organic acid (90%) in all raspberry cultivars and the Rubi cultivar showed the highest total non-volatile organic acids content (2003 mg/100 g f.w.). Hunter color CIE values were also determined, showing that Rubi was the reddest raspberry cultivar. Received: 28 May 1998     

Optimised processing technique for colour- and cloud-stable plum juices and stability of bioactive substances

Plum (Prunus domestica) juices were produced from different cultivars with optimised processing technology. High results for dry mass (19–21 °Brix), sugar-free extract (75–105 g/L), total acidity (6–11 g/L), sorbitol (27–52 g/L) and minerals were found. Secondary plant metabolites were present in high amounts as well. By means of HPLC/PDA 1465–2590 mg/L of colourless polyphenols were found. Neochlorogenic acid, chlorogenic acid, (+)-catechin and (−)-epicatechin were the predominant polyphenols. Total anthocyanins were found in amounts of 43–168 mg/L in the fresh juices. They were identified as cyanidin-3-glucoside, cyanidin-3-rutinoside, peonidin-3-glucoside and peonidin-3-rutinoside. A fast decline of the anthocyanin concentrations could be observed during a 6 months storage at 20 °C, which changed the red colour of the juices drastically. Beside high polyphenol concentrations, high antioxidant capacities of the juices were determined as well. The applied processing technology allowed the production of cloud-stable plum nectars.     

Fruit quality and bioactive compounds relevant to human health of sweet cherry (Prunus avium L.) cultivars grown in Italy

The fruit quality characteristics, phenolic compounds and antioxidant capacities of 24 sweet cherry (Prunus avium L.) cultivars grown on the mountainsides of the Etna volcano (Sicily, Italy) were evaluated. High-performance liquid chromatographic methods were used to identify and quantify sugars, organic acids and phenolics. A total of seven phenolic compounds were characterised as hydroxycinnamic acid derivatives (neochlorogenic acid, p-coumaroylquinic acid and chlorogenic acid) and anthocyanins (cyanidin 3-glucoside, cyanidin 3-rutinoside, pelargonidin 3-rutinoside and peonidin 3-rutinoside). The total anthocyanin content ranged from 6.21 to 94.20 mg cyanidin 3-glucoside equivalents/100 g fresh weight (FW), while the total phenol content ranged from 84.96 to 162.21 mg gallic acid equivalents/100 g FW. The oxygen radical absorbance capacity (ORAC) assay indicated that fruit of all genotypes possessed considerable antioxidant activity. The high level of phenolic compounds and antioxidant capacity of some sweet cherry fruits implied that they might be sources of bioactive compounds that are relevant to human health.     

Lingonberry (Vaccinium vitis-idaea L.) grown in the Pacific Northwest of North America: Anthocyanin and free amino acid composition

Lingonberries and their products are popular and generally accessible in Europe, though in the US they are uncommon and considered a minor berry/fruit crop. The on-going interest in potential health benefits from berry consumption has heightened interest in broadening the selection of berry/fruit crops in the US. This study measured total phenolics, total tannins, complete anthocyanin content, and total (and individual) free amino acid composition for each of five lingonberry cultivars. Cultivars Ida, Koralle, Linnea, Sanna, and Sussi were grown in Oregon, USA, and had only been evaluated previously for their horticultural traits. All five cultivars contained the three anticipated anthocyanins (by HPLC): cyanidin-3-galactoside (main anthocyanin found in these berries), cyanidin-3-glucoside, and cyanidin-3-arabinoside. These lingonberries’ total anthocyanin content ranged from 27.4 (‘Linnea’) to 52.6 (‘Ida’) mg/100 g fw. They contained 22 free amino acids (FFAs) and total FAAs ranged from 28.92 (‘Sanna’) to 70.38 (‘Koralle’) mg/100 g fw. Asparagine (ASN) was the leading FAA (22–34% of the total FAAs) for all five cultivars. This is the first report on lingonberry FAA content.     

Anthocyanin Pigment Composition of Blackberries

ABSTRACT: The anthocyanin pigments in 51 blackberry samples were isolated and characterized by highperformance liquid chromatography (HPLC) with UV-visible detection, electrospray mass spectroscopy (ESMS), and hydrolysis and saponification reactions. Five pigments were detected: cyanidin-3-glucoside, cyanidin-3-rutinoside, cyanidin-3-xyloside, cyanidin-3-glucoside acylated with malonic acid, and an unidentified acylated derivative of cyanidin-3-glucoside. Four different quantitative patterns were evident among the cultivars and selections. Total anthocyanin content ranged from 70.3 to 201 mg/100 g with a mean of 137 mg/100 g. Ten commercial blackberry juice concentrate samples were analyzed. Total anthocyanin ranged from 12.3 to 107 mg/100 g on a single-strength juice basis, and 1 sample had an atypical profile.     

Phenolic compound content of fresh and dried figs (Ficus carica L.)

Eighteen fig cultivars commonly grown in the south-eastern Spain were collected in two crops (spring and summer) and their polyphenolic profile was determined. Fruit from the first crop generally showed higher phenolic values than those from the second. Higher concentrations of total phenolics were found in skin than in flesh. LC–UV-DAD/ESI-MSⁿ analysis of the figs pointed to a high anthocyanin content, mainly cyanidin-3-rutinoside, flavonols such as quercetin-rutinoside, phenolic acids such as chlorogenic acid and flavones like luteolin 6C-hexose-8C-pentose and apigenin-rutinoside. The c-glycosides have not been previously described in figs. Moreover, to compare intact proanthocyanidins and proanthocyanin cleavage products, an acid-catalysis was made in the presence of phloroglucinol (phloroglucinolysis) and the mean degree of polymerisation was calculated. Finally, three cultivars of dried figs were also studied.     

Determination of anthocyanins in four Croatian cultivars of sour cherries (Prunus cerasus)

The anthocyanins were characterised and quantified in four cultivars of sour cherries by means of high-performance liquid chromatography and mass spectrometry. Column chromatography on XAD-2 Amberlite was the method used for isolation of anthocyanins. A diode-array detector was employed. For anthocyanins determination was chosen wavelength of 518 nm. Cyanidin-3-glucosylrutinoside and cyanidin-3-rutinoside were major anthocyanins. The total anthocyanin content in sour cherries, (expressed as cyanidin-3-glucosylrutinoside) varied from 2.7 to 28.0 mg/100 g of fresh weight, with the highest content being observed in Oblainska cultivar.     

Quality changes during ripening of plums (Prunus domestica L.)

Quality changes during fruit ripening after the appearance of fruit colour of four Prunus domestica L. plum cultivars, ‘Jojo’, ‘Valor’, ‘Čačanska rodna’ and ‘Čačanska najbolja’, were investigated during 25 or 33 day periods. Fruit samples were analyzed for fruit weight, firmness, soluble solids content, fruit colour, content of sugars (glucose, fructose, sorbitol and sucrose), organic acids (malic, fumaric and shikimic acids), phenolics (neochlorogenic acid, p-coumaroylquinic acid, chlorogenic acid and rutin) and anthocyanins (cyanidin-3-rutinoside and peonidin-3-rutinoside). Ripening resulted in statistically increased fruit weight and soluble solids, decreased fruit firmness, darker colour of fruits, increased concentration of total sugars, decreased concentration of total acids, and increased concentration of anthocyanins. There was no influence of ripening on the content of phenols. The results show significant influences of cultivar on fruit weight, soluble solids content, firmness, fruit colour, concentration of total acids, SUAC index, concentration of total phenols and anthocyanins in European plums.     

Polyphenolic profile and antioxidant activity of five apple cultivars grown under organic and conventional agricultural practices

The polyphenols and total antioxidant activities of five apple cultivars, grown by organic and conventional agricultural methods in neighbouring farms, were determined and compared. Total polyphenols in the whole fruit and in the peel were determined by the Folin-Ciocalteu method, and the total antioxidant activity was determined by three established methods, diphenyl picrylhydrazyl (DPPH), azinobis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and ferric reducing/antioxidant power (FRAP). Polyphenolic content for the whole fruit was in the range of 80–196 and for the peel 165–400 (mg Gallic Acid Equivalent (GAE)/100 g fresh weight) for both types of agricultural practices. Antioxidant activities of fruit extracts were also relatively similar and well correlated to their polyphenolic content. High-performance liquid chromatography (HPLC) analysis of the most important polyphenolics (chlorogenic acid, catechin, epicatechin, procyanidin B1 and B2, cyaniding 3-galactoside, phloridzin, quercetin 3-galactoside and quercetin 3-arabinoside) also showed that concentrations do not differentiate significantly between the organic and conventional apples. Statistical significance of differences in antioxidant activities among the same cultivars was relatively small (flesh + peel or peel only) for both types of apples. These results indicate that organic apples do not present higher antioxidant or nutritional value compared with conventionally grown ones, as far as polyphenolic content and total antioxidant activities are concerned.     

野生蓝果忍冬多酚鉴定及其抗氧化、降血糖活性

目的：对野生蓝果忍冬进行多酚成分鉴定和抗氧化、抗淀粉酶活性研究。方法：以我国小兴安岭地区的野生蓝果忍冬果实为材料,通过固相萃取技术分离纯化得到花色苷和非花色苷分离液,测定分离液中的化学成分,并测定多酚粗提液中总酚和总花色苷含量,抗氧化活性（1,1-二苯基-2-三硝基苯肼自由基清除能力、铁离子还原能力、2,2’-联氮-双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基清除能力、氧自由基吸收能力）,抗α-淀粉酶、α-葡萄糖苷酶活性和脂肪酶活性。结果：野生蓝果忍冬果实中含有丰富的花色苷和非花色苷多酚,尤其是矢车菊素-3-葡萄糖苷和山柰酚-3-芸香糖苷含量很高;蓝果忍冬多酚粗提液中总酚含量为82.7 mg/100 g,总花色苷含量为49.8 mg/100 g,具有较强的抗氧化活性;对α-淀粉酶、α-葡萄糖苷酶活性的半抑制质量浓度分别为0.39 mg/mL和0.933 mg/mL,对脂肪酶活性的半抑制质量浓度为12.31 mg/mL,说明蓝果忍冬可以作为一种有前景的淀粉酶抑制剂。     

Comparative study of phenolic compounds and antioxidant activity in different species of cherries

A new spectrometric method ultra performance liquid chromatography-tandem mass spectrometric with high precision and rapid analysis was developed to separate 17 phenolic compounds. Different species of cherries, including 10 sweet cherry (Prunus avium L.) cultivars, a tart cherry (P. cerasus L.) rootstock (CAB), and a hybrid rootstock 'Colt' (P. avium × P. pseudocerasus), were analyzed for phenolics contents by this method. The results showed that significant differences were observed among the phenolic compound contents in different cherry species. In 10 sweet cherry cultivars, the contents of neochlorogenic acid and cyanidin-3O-rutinoside were much higher in red-colored fruits (for example, 64.60 and 44.50 mg/100 g fresh weight in Burlat, respectively) than those in bicolored ones. Principal component analysis revealed that cyanidin-3O-rutinoside was an effective index for grouping the cultivars with similar species and fruit colors. Moreover, there were strong positive correlations between phenolics content and antioxidant activity, which was higher in red-colored cherries.     

Optimization of microwave-assisted extraction of anthocyanins in red raspberries and identification of anthocyanin of extracts using high-performance liquid chromatography – mass spectrometry

Anthocyanins (Acys) are naturally occurring compounds that impart color to fruit, vegetables, and plants. The extraction of Acys from red raspberry (Rubus idaeus L. var. Heritage) by microwave-assisted process (MAP) was studied. A central composite rotate design (CCRD) was used to obtain the optimal conditions of microwave-assisted extraction (MAE), and the effects of operating conditions such as the ratio of solvents to materials, microwave power and extraction time on the extraction yield of Acys were studied through response surface methodology (RSM). The optimized conditions of MAE were ratio of solvents to materials 4:1 (ml/g), extraction time 12 min, and microwave power 366 W. Under these conditions 43.42 mg of Acys from 100 g of fresh fruits (T _Acy, expressed as cyanidin-3-glucoside), approximately 98.33% of the total red pigments, could be obtained by MAE. The Acys compositions of extracts were identified by high-performance liquid chromatography – mass spectrometry (HPLC-MS), 12 kinds of Acys had been detected and 8 kinds of Acys were characterized. Result indicated that cyanidin-3-sophoroside, cyanidin-3-(2 ^G -glucosylrutinoside), cyanidin-3-sambubioside, cyanidin-3-rutinoside, cyanidin-3-xylosylrutinoside, cyanidin-3-(2 ^G -glucosylrutinoside), and cyanidin-3-rutinoside were main components in extracts. In addition, in comparison with the conventional solvent extraction, MAE is more efficient and rapid to extract Acys from red raspberry, due to the strong disruption of fruit tissue structure under microwave irradiation, which had been observed with the scanning electron microscopepy (SEM). However, the Acys compositions in extracts by both the methods were similar, which were investigated using HPLC profile.     

Comparative study of primary and secondary metabolites in apricot (Prunus armeniaca L.) cultivars

BACKGROUND: Apricot is one of the most popular Prunus species grown in the temperate zone and great variation in the contents of primary and secondary metabolites has been reported in its fruit. However, little is known on the phenolic profiles and antioxidant potential of important local cultivars. The objective of the study was to identify valuable parental lines for breeding of cultivars with high health‐promoting effects. RESULTS: Primary metabolites (sugars, organic acids) and secondary metabolites (phenolics) were quantified by high‐performance liquid chromatography–mass spectrometry (HPLC/MS) in fruit of 13 apricot cultivars: total sugars ranged from 59.2 to 212.5 g kg^?1 fresh weight (FW) and total organic acids from 4.2 to 20.8 g kg^?1 FW. Four hydroxycinnamic acids and three flavonols were quantified; their content was significantly higher in skin compared to pulp. Similarly, antioxidative potential was significantly higher in skin and ranged from 125.4 to 726.5 mg ascorbic acid equivalents kg^?1 FW. A positive correlation between total phenolic content and antioxidant potential was determined. Multivariate analysis subdivided the cultivars into five major groups, mostly influenced by the content of individual and total phenolics as well as the antioxidant potential of apricot skin and pulp. CONCLUSION: In regard to high phenolic content in connection with antioxidant potential, three cultivars stand out as the richest in analyzed quality parameters: ‘Mula Sadik’, ‘Bergeron’ and ‘Chuan Zhi Hong’. They also contained high amounts of primary metabolites and should be promoted for further planting and included in apricot breeding programs. Copyright © 2010 Society of Chemical Industry     

Characterization of Anthocyanins in Fruit of Kadsura coccinea (Lem.) A.C. Smith by UPLC/Q-TOF-MS Analysis and Evaluation of Stability of the Major Anthocyanins

Anthocyanins in fruit of Kadsura coccinea (Lem.) A.C. Smith were identified by ultra-performance liquid chromatography–quadrupole–time of flight mass spectrometry analysis. Contents of anthocyanins in K. coccinea fruit were determined by HPLC analysis. Colour stability, thermal degradation and antioxidant activity of purified cyanidin-3-xylosylrutinoside were further analysed. As a result, four anthocyanins were identified as delphinidin-3-xylosylrutinoside (1), cyanidin-3-glucosylrutinoside (2), cyanidin-3-xylosylrutinoside (3), and cyanidin-3-rutinoside (4). The individual content of anthocyanins identified was 4.36?±?0.06, 3.01?±?0.03, 49.92?±?0.76 and 2.33?±?0.04 mg/500 g deseeded K. coccinea fruit, respectively. Cyanidin-3-xylosylrutinoside was stable at pH around 1. Moreover, cyanidin-3-xylosylrutinoside was very sensitive to temperature change and it was even unstable at room temperature. Cyanidin-3-xylosylrutinoside tended to degrade into cyanidin-3-rutinoside, cyanidin-sambubioside and cyanidin at high temperatures. Cyanidin-3-xylosylrutinoside exhibited much better 1,1-diphenyl-2-picrylhydrazyl and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical scavenging activities than those of butylated hydroxytoluene. However, cyanidin-3-xylosylrutinoside and other three anthocyanin components contributed little to the antioxidant activity of K. coccinea fruit.     

HPLC法测定桃不同品种果实中花色苷组成和含量

目的： 用高效液相色谱法测定桃不同品种果实中花色苷组成和含量。方法： 色谱柱为C1 8柱(150mm×4.6mm,5μm),流动相为乙腈：4%磷酸溶液(V/V=12:88,pH2),检测波长520nm,流速0.8mL/min,进样量20μL,柱温30℃。结果：桃3个品种果实中主要含有1种花色苷：花青素-3-葡萄糖苷,花青素-3-葡萄糖苷在0.4～100μg/mL内线性关系良好(R=0.9992),低、中、高质量浓度的平均回收率为85.7%、89.4%、91.2%。红、黄、白肉桃果肉中的花青素-3-葡萄糖含量分别为103.7、30.6、21.5mg/kg。结论：本方法操作简单,结果准确可靠,重复性好,可作为桃果实中花青素-3-葡萄糖苷的定量分析方法。     

High performance liquid chromatography analysis of anthocyanins in bilberries (Vaccinium myrtillus L.), blueberries (Vaccinium corymbosum L.), and corresponding juices

In the present study the anthocyanin content of commercially available bilberry juices and fresh fruits were quantified by using 15 authentic anthocyanin standards via high performance liquid chromatography with an ultra-violet detector (HPLC-UV/VIS). Delphinidin-3-O-glucopyranoside, delphinidin-3-O-galactopyranoside, and cyanidin-3-O-arabinopyranoside were the major anthocyanins found in juices, nectar, and fresh bilberries. In contrast, fresh blueberries had higher concentrations of malvidin-3-O-arabinopyranoside and petunidin-3-O-galactopyranoside. Up to 438 mg anthocyanins per 100 g fresh weight (2762 mg/100 g dry weight (DW)) were detected in blueberries from various sources, whereas bilberries contained a maximum of 1017 mg anthocyanins per 100 g fresh weight (7465 mg/100 g DW). Commercially available bilberry and blueberry juices (n= 9) as well as nectars (n= 4) were also analyzed. Anthocyanin concentrations of juices (1610 mg/L to 5963 mg/L) and nectar from bilberries (656 mg/L to 1529 mg/L) were higher than those of blueberry juices (417 mg/L) and nectar (258 mg/L to 386 mg/L). We conclude that using several authentic anthocyanin references to quantify anthocyanin contents indicated them to be up to 53% and 64% higher in fresh bilberries and blueberries, respectively, than previously reported using cyanidin-3-O-glucoside. This study has also demonstrated that commercially available juices produced from bilberries contain much higher anthocyanin concentrations than those from blueberries. PRACTICAL APPLICATION: We have investigated the contents of a special class of antioxidants, namely anthocyanins in blueberry and billberry fruits and juices commercially available in Germany. To achieve reliable data we have used authentic standards for the first time. We think that our results are important in the field of nutritional intake of this important class of polyphenols and fruit juice companies get a closer insight in the occurrence of these antioxidants in market samples to be used in food composition databases and for nutritional survey.     

Analytical characterization and the impact of ageing on anthocyanin composition and degradation in juices from five sour cherry cultivars

Five different sour cherry cultivars were processed to sour cherry juices in order to analyze their composition and the impact of storage on anthocyanin composition. Regarding primary juice parameters all cultivars showed high dry masses (13.8–17.2° Brix) and sugar free extracts (59.4–74.3 g/L). Anthocyanins were identified as cyanidin-3-(2^G-glucosylrutinoside), cyanidin-3-(2^G-xylosylrutinoside), cyanidin-3-sophoroside, cyanidin-3-glucoside, cyanidin-3-rutinoside, and peonidin-3-rutinoside. Besides the original anthocyanins, 5-carboxypyrano-anthocyanins occurred as new reaction products during storage. HPLC-MSⁿ experiments revealed, that a further ageing pigment was probably resulting from the reaction of an original anthocyanin with acetaldehyde. A significant decline of the original anthocyanin concentrations could be observed (70–75%) during 6 months of storage at 20 °C. Degradation kinetics of anthocyanins were calculated taking the reaction rate as first order. Colourless polyphenol concentrations and antioxidant capacities (TEAC) of the juices were not affected during storage.     

The effects of six phenolic acids and tannic acid on colour stability and the anthocyanin content of mulberry juice during refrigerated storage

Six phenolic acids and tannic acid are added to fresh mulberry juice to enhance the colour. Effects of these copigments on colour stability and the anthocyanin content of mulberry juice are investigated while being stored at 4 °C for 30 days. Results suggest that the addition of the copigments markedly increases the maximum absorbance of mulberry juice. Cyanidin-3-glucoside is the major anthocyanin, followed by cyanidin-3-rutinoside. Furthermore, the retention rate of the total anthocyanin, cyanidin-3-rutinoside and cyanidin-3-rutinoside increases after the addition of all copigments. The effect of ferulic acid on the stability of total anthocyanin, cyanidin-3-glucoside and cyanidin-3-rutinoside in mulberry juice is the best, 89.52%, 86.90% and 94.46% respectively (66.56%, 62.58% and 73.60% in the control group). Researchers recommend the addition of ferulic acid, p-coumaric acid and caffeic acid – all of which belong to the hydroxycinnamic acid group, to the mulberry juice to enhance the colour stability and anthocyanin content.     

Quantitative changes in anthocyanin pigments of lychee fruit during refrigerated storage

Changes in cyanidin-3-glucoside, cyanidin-3-rutinoside, malvidin-3-acetylglucoside, total anthocyanins, percent polymeric color and browning indexes in stored lychee (Litchi chinensis Sonn.) fruits were determined. Total anthocyanin content declined from 1·77 to 0·73 mg/g fresh weight, and individual anthocyanins also decreased throughout the 48-day storage period. Decline in anthocyanins was accompanied by an increase in browning. Polymeric pigments also gradually increased from 20·9 to 53%.