HPLC chromatogram of anthocyanin

Figure 8. HPLC chromatogram of anthocyanin derivatives of Merlot wine corresponding to 1997 vintage.

Figure 20. HPLC chromatograms of anthocyanins (UV detection 525 nm, private communication).

Fig. 2.71. HPLC chromatogram of the neutral (a) and acidic fractions (b) and the acid-catalysed hydrolysed product of freshly squeezed cranberry juice (c) at 280 nnm. Peaks in a 1 = ( + )-cate-chin 2 = myicetin 3 = quercetin (added as internal standard). Peaks in b 1 = anthocyanin derivative I 2 = benzoic acid 3 = p-anisic acid 4 = quercetin (added as internal standard). Peaks in c 1 = ( + )-catechin 2 = anthocyanin derivative I 3 = anthocyanin derivative II 4 = benzoic acid 5 = anthocyanin derivative III 6 = p-anisic acid 7 = myricetin 8 = quercetin. Reprinted with permission from H. Chen et al. [188].

Fig. 4.2 HPLC-DAD chromatograms of anthocyanins from soluble and insoluble extracts of black, red-brown, and brown soybean seed coats at 520 nm. (a, d) Black (Clark), (b, e) red-brown (Mil), and (c, f) brown (MlOO) seed coats, (a and b) Pulverized fresh seed coats extracted with 80% methanol in water, (d-f) Insoluble pulverized seed coat fraction extracted with 1-butanol/HCl (19 1) 1% SDS. Compound identifications were based on comparison of retention times and absorption spectra to authentic standards. Peak 1, unknown peak 2, delphinidin-3-O-galactoside peak 3, delphinidin-3-O-glucoside peak 4, cyanidin-3-O-galactoside peak 5, cyanidin-3-O-glucoside peak 6, petunidin-3-O-glucoside peak 7, pelargonidin-3-O-glucoside peak 8, peonidin-3-O-glucoside and peak 9, malvidin-3-O-glucoside...

Figure F1.3.4 HPLC chromatogram of concord grape (solid line) and strawberry (dashed line) anthocyanins. Peaks identified on figure.

Figure F1.3.6 HPLC chromatograms of radish anthocyanin extract before (solid line) and after (dashed line) saponification. Peak 1 pelargonidin-3-sophoroside-5-g ucoside Peaks 2, 3, 4, 5 pelargonidin-3-sophoroside-5-glucoside acylated with />coumaric (2), ferrulic (3), p-couniaric and malonic (4), or ferrulic and malonic acid (5), respectively. Note that the saponified sample has only pelargonidin-3-sophoroside-5-glucoside.

Figure 11.3.12 HPLC chromatograms of polyphenolics in Concord grape extract detected at 280 nm. (A) All polyphenolics, including anthocyanins. (B) Nonanthocyanin polyphenolics after fractionation. Peak identification 1, cis-caftaric acid 2, frans-caftaric acid 3, procyanidin B3 4, c/s-coutaric acid 5, frans-coutaric acid 6, epicatechin 7, quercetin galactoside 8, quercetin glucoside. Reproduced from Oszmianski and Lee (1990) with permission from the American Society for Enology and Viticulture.

Fig. (8). HPLC chromatogram of red lettuce anthocyanins and other phenols (1) caffeoyltartaric acid, (2) chlorogenic acid, (3) dicaffeoyltartaric acid, (4) quercetin 7-glucoside 3-(6 -malonylglucoside), (5) dicaffeoylquinic acid dsochlorogenic acid), (6) quercetin 3-glucuronido, (7) quercetin 3-glucosido, (8) quercetin 3-malonylglucoside, and (9) cyanidin 3-malonylglucoside. HPLC conditions RP Cjg column (12 x 0.4 cm particle size 5 pm). Mobile phase acidified water (5% formic acid) (A) and methanol (B). Gradient 0 min- 5% B, 25 min- 40% B. Flow rate I mL/min.

Fig. 10 shows HPLC chromatograms of red onion phenolics. Four anthocyanins are visible in the 520 nm chromatogram and three principal flavonoids can be clearly observed in the 260 nm chromatogram. Hydroxycinnamic acid or benzoic acid derivatives cannot be observed in any significant quantity in the chromatograms. 

Fig. 6.34. HPLC chromatogram of an anthocyanin solution extracted from Merlot skins as monoglncosides 20, delphinidin 21, cyanidin 22, petunidin 23, peonidin 24, malvidin 25, 26, 27, 28, 29, 30, acylated anthocyanins (Galvin, 1993)...

Based on peak size of HPLC chromatogram or percent above 10. Only anthocyanins exceeding 1% listed. 

The spectra were recorded in the positive-ion mode in the range of m/z 120-1 500. Some chromatograms illustrating the effect of aldehydes on the interaction of mv3gl and B2-3 -gallate are shown in Fig. 2.120. The chromatograms demonstrate that different aldehydes influence differently the formation of anthocyanin-flavanol pigments. The results of HPLC-MS measurements are compiled in Table 2.94. Because free aldehydes display an unpleasant aroma in Port wine these reactions may improve the quality of wines and contribute to the colour formation [266],... 

Either Basic Protocol 1 or the Alternate Protocol should first be conducted in analyzing an unknown sample. Because of its ease and simplicity, sample preparation of anthocyanins and their HPLC separation on silica Cl8 columns (see Basic Protocol 1) is usually the preferred choice, unless the presence of acylated anthocyanins is anticipated, in which case the protocol described for acylated anthocyanins is used (see Alternate Protocol). If the anthocyanin profile is inconsistent with previously published chromatograms, or if there are extraneous unidentified peaks, then simplification is recommended (see Basic Protocol 3). Acid hydrolysis will simplify the chroma-... 

Figure FI. 3.4 shows HPLC chromatograms for anthocyanidins generated from acid hydrolysis of concord grape and strawberry juices. Extraneous peaks may be present because of incomplete hydrolysis, and degradation and polymerization of the labile aglycons even more of a problem. For acylated anthocyanins, higher yields of anthocyanidins will be achieved if the sample is first saponified (see Basic Protocol 3) and then subjected to acid hydrolysis (see Basic Protocol 2).

HPLC chromatograms with peak assignments are given for several commodities along with applications of anthocyanin analyses for determining authenticity of fruit juices. 

Two predominant phenolic compounds (neochlorogenic and chlorogenic acids) in prunes and prune juice can be analyzed by reversed-phase HPLC with diode array detection along with other phenolic compounds (65). Phenolic compounds were extracted from prunes with methanol and aqueous 80% methanol and analyzed by HPLC. Ternary-gradient elution (a) 50 mM NaH4H2P04, pH 2.6, (b) 80% acetonitrile/20% (a), and (c) 200 mM phosphoric acid, pH 1.5, was employed for an 80-min run time. Four wavelengths were monitored for quantitation 280 nm for catechins and benzoic acids, 316 nm for hydroxycinnamates, 365 nm for flavonols, and 520 nm for anthocyanins. Phenolic analysis of pitted prune extract is presented in an HPLC chromatogram in Fig. 9, which is based on work done by Donovan and Waterhouse (65). 

Since the work of Manley and Shubiak (182), who were the first to apply HPLC to anthocyanin analysis, numerous HPLC techniques have been developed for the separation and quantification of anthocyanins and anthocyanidins. Nowadays HPLC has become the method of choice, because it offers the advantage that it is a rapid, sensitive, and quantitative method. For the peak identification and quantitative evaluation of chromatograms, the use of pure anthocyanin standards is recommended however, only a limited, but constantly increasing, number of substances is avail-... 

FIGURE 3.11 HPLC chromatogram (a) and ESMS spectrum (b) of anthocyanins in highbush blueberry (Vaccinium corymbosum var. Bluecrop) extracts. The anthocyanins were isolated by solid-phase extraction of C-18 cartridges with acidified methanol—cartridges had previously been washed with acidified water and by ethyl acetate. Peak identities for both the HPLC chromatogram and ESMS spectrum correspond to numbers in Table 3.5. [Source Skrede et al. (2000a), with permission.]... 

Table 2.4 HPLC gradient program used for analysis of anthocyanins in grape skins extract by C18 (250 x 4mm, 5 pm) column (chromatogram in Figure 2.14) (flow rate 0.5mL/min).

Figure 10. Proportion of monomeric anthocyanin pigment (Mv, Afv-3-(acetyl)glucoside, Mv-3-(p-coumaryl)-glucoside, vitisin A, acetyl-vitisin A, p-coumaryl-vitisin A) to polymeric pigment (unidentifiedpigments in the hump) as defined by the area under the HPLC chromatogram at 520 nm. The wine was made from Shiraz grapes and was measured directly after fermentation and then at 6 months and at 12 months of age.

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