Effect of in vitro gastrointestinal digestion on the composition and bioactivity of anthocyanins in the fruits of cultivated Lycium ruthenicum Murray

Figures & data

Table 1. Identification and the contents of individual anthocyanins and total anthocyanins before and after in vitro gastrointestinal digestion.^a

Tabla 1. Identificación y contenido de antocianinas individuales y antocianinas totales antes y después de la digestión gastrointestinal in vitro.^a

					Concent of anthocyanins (μg/g)			Proportion of anthocyanins (%)
Peak no.	TR (min)	Molecular (M^+)	Fragment (m/z)	Identification	Hedong	Delingha	Nomuhong	Hedong	Delingha	Nomuhong
Before
1	24.087	317.1277	-	Unknown	1560.96 ± 56.82^a	1120.72 ± 42.05^c	1283.48 ± 74.11^b	9.77 ± 0.17^a	7.67 ± 0.16^c	8.45 ± 0.41^b
2	41.530	641.5050	479.1287/317.1277	Petunidin-3-O-galactoside-5-O-glucoside	513.82 ± 30.89^c	670.81 ± 27.27^a	617.89 ± 13.21^b	3.22 ± 0.18^c	4.59 ± 0.17^a	4.07 ± 0.11^b
	41.530	633.5297	465.3024/303.2303	Unknown
3	47.273	641.5050	479.1287/317.1277	Petunidin-3-O-glucoside-5-O-glucoside	151.66 ± 25.73^a	137.45 ± 7.78^a	143.05 ± 15.71^a	0.95 ± 0.16^a	0.94 ± 0.04^a	0.94 ± 0.10^a
4	50.903	919.7361	757.3324/627.0011/465.0068/303.0531	Delphinidin-3-O-rutinoside (cis-p-coumaroyl)-5-O-glucoside	109.25 ± 5.71^b	122.22 ± 3.76^b	138.15 ± 8.94^a	0.68 ± 0.04^b	0.84 ± 0.04^a	0.91 ± 0.07^a
5	57.023	919.7361	757.3324/627.0011/465.0068/303.0531	Delphinidin-3-O-rutinoside (trans-p-coumaroyl)-5-O-glucoside	657.97 ± 31.61^a	472.69.±117.60^b	424.57 ± 22.64^b	4.12 ± 0.17^a	3.24 ± 0.15^b	2.80 ± 0.14^c
6	57.520	949.7525	786.9683/641.5133/478.9793/317.0045	Petunidin-3-O-rutinoside (caffeoyl)-5-O-glucoside	132.12 ± 13.71^b	185.88 ± 28.62^a	190.16 ± 22.89^a	0.83 ± 0.09^b	1.27 ± 0.21^a	1.25 ± 0.15^a
7	64.950	933.6037	771.4425/641.5133/478.4011/317.0617	Pentunidin-3-O-rutinoside (cis-p-coumaroyl)-5-O-glucoside	863.75 ± 30.02^b	817.92 ± 18.73^b	847.80 ± 11.75^a,b	5.41 ± 0.19^a	5.60 ± 0.05^a	5.58 ± 0.09^a
8	67.403	933.6037	771.4425/641.5133/478.4011/317.0617	Pentunidin-3-O-rutinoside (trans-p-coumaroyl)-5-O-glucoside	111239.49 ± 241.57^a	10367.60 ± 233.38^b	10769.45 ± 99.46^b	70.38 ± 0.29^b	70.95 ± 0.30^a	70.91 ± 0.12^a
9	72.143	757.5598	641.0078/479.2581/317.0617	Pentunidin-3-O-glucoside (maloyl)-5-O-glucoside	97.40 ± 7.25^a	92.61 ± 3.22^a	94.98 ± 4.90^a	0.61 ± 0.04^a	0.63 ± 0.01^a	0.63 ± 0.03^a
10	73.400	963.8261	786.9683/641.5133/479.0000/317.0017	Pentunidin-3-O-glucoside (feruloyl)-5-O-glucoside	129.132 ± 8.70^b	142.01 ± 4.82^b	163.52 ± 11.51^a	0.81 ± 0.07^b	0.97 ± 0.03^a	1.08 ± 0.09^a
11	77.083	947.2676	785.1089/493.7355/331.2977	Malvidin-3-O-rutinoside (cis-p-coumaroyl)-5-O-glucoside	244.04 ± 3.69^c	253.78 ± 2.73^b	270.72 ± 6.39^a	1.53 ± 0.05^b	1.74 ± 0.05^a	1.78 ± 0.06^a
12	79.110	977.3944	771.8603/478.8520/317.0045	Unknown	187.87 ± 7.25^a	127.93 ± 8.85^c	152.11 ± 8.11^b	1.18 ± 0.02^a	0.88 ± 0.06^c	1.00 ± 0.06^b
13	82.328	917.7199	642.5133/479.6006/317.2415	Unknown	82.43 ± 3.12^c	99.57 ± 2.93^a	92.05 ± 1.64^b	0.52 ± 0.03^c	0.68 ± 0.03^a	0.61 ± 0.02^b
Total anthocyanins					15969.89 ± 324.33^a	14611.19 ± 281.33^c	15187.92 ± 156.57^b
After
1	9.950	303.0472	-	Delphinidin	541.26 ± 6.14^a	465.36 ± 5.76^c	520.42 ± 4.88^b	3.90 ± 0.02^a	4.05 ± 0.14^a	3.97 ± 0.08^a
2	10.707	287.0696	-	Cyanidin	26.34 ± 0.92^a	22.31 ± 0.65^b	23.58 ± 0.78^b	0.19 ± 0.01^a	0.19 ± 0.00^a	0.18 ± 0.00^a
3	12.027	317.0646	-	Petunidin	13100.72 ± 107.16^a	11083.25 ± 92.40^c	12382.18 ± 274.77^b	94.52 ± 0.05^a	95.74 ± 2.27^a	94.50 ± 0.12^a
4	12.937	271.0219	-	Pelargonidin	22.44 ± 1.01^a	18.99 ± 0.84^b	20.41 ± 1.18^a,b	0.16 ± 0.01^a	0.16 ± 0.01^a	0.16 ± 0.01^a
5	14.737	331.0789	-	Malvidin	169.86 ± 4.43^a	145.17 ± 2.71^c	155.78 ± 6.11^b	1.23 ± 0.03^a	1.25 ± 0.03^a	1.19 ± 0.06^a
Total anthocyanins					13860.62 ± 117.29^a	11579.96 ± 201.68^c	13102.37 ± 273.87^b

^a Data with the same superscript in the same row are not significantly different (P> 0.05).

^aLos datos con el mismo superíndice en la misma fila no son significativamente diferentes (P> 0.05).

Figure 1. HPLC chromatograms of CLM anthocyanins in the samples before (a) and after (b) in vitro gastrointestinal digestion. The detection wavelength is 525 nm, and the peak names are listed in Table 1.

Figura 1. Cromatogramas de HPLC de antocianinas CLM en las muestras antes (a) y después (b) de la digestión gastrointestinal in vitro. La longitud de la onda de detección es de 525 nm; los nombres de los picos se enumeran en la Tabla 1.

Figure 2. Chemical structures of CLM anthocyanins.

Figura 2. Estructuras químicas de las antocianinas CLM.

Table 2. Loadings of the features in the first three principal components for CLM anthocyanins before in vitro gastrointestinal digestion.^a

Tabla 2. Cargas de las características en los primeros tres componentes principales de las antocianinas CLM antes de la digestión gastrointestinal in vitro.^a

Variable	PC 1	PC 2	PC 3
Peak 1	−0.946	0.002	0.269
Peak 2	0.925	0.114	−0.342
Peak 3	−0.020	0.875	−0.013
Peak 4	0.858	−0.061	0.480
Peak 5	−0.893	0.018	−0.323
Peak 6	0.848	−0.193	−0.136
Peak 7	0.756	0.520	0.260
Peak 8	0.711	−0.468	−0.030
Peak 9	0.413	0.807	0.079
Peak 10	0.861	−0.237	0.434
Peak 11	0.941	−0.116	0.173
Peak 12	−0.862	−0.026	0.463
Peak 13	0.901	0.050	−0.399

^aPeak names refer to Table 1.

^aLos nombres de los picos se refieren a la Tabla 1.

Figure 3. PCA two-dimensional plots of the anthocyanin composition in CLM before in vitro gastrointestinal digestion, which shows the grouping of the samples according to the cultivating location.

Figura 3. Gráficos bidimensionales de PCA de la composición de antocianina en CLM antes de la digestión gastrointestinal in vitro, que exhiben el agrupamiento de las muestras según la ubicación del cultivo.

Table 3. The actual weight and the purity of CLM anthocyanins collected form 1 g samples before and after gastrointestinal digestion.^a,b.

Tabla 3. El peso real y la pureza de las antocianinas CLM recolectadas forman muestras de 1 g antes y después de la digestión gastrointestinal.^{a, b.}

		Hedong	Delingha	Nomuhong
Weight (μg)	Before	16409.67 ± 397.38^a	15039.83 ± 539.49^c	15651.25 ± 455.38^b
	After	14071.70 ± 290.34^a	11861.07 ± 356.27^c	13391.63 ± 335.59^b
Purity (%)	Before	97.32^a	97.15^a	97.04^a
	After	98.05^a	97.63^a	97.84^a

^aThe purity (%) = (the content of CLM anthocyanins obtained by quantitative analysis (Table 1))/(the actual weight of collected CLM anthocyanins) × 100.

^bData with the same superscript in the same row are not significantly different (P> 0.05).

^aLa pureza (%) = (el contenido de antocianinas CLM obtenido por análisis cuantitativo (Tabla 1))/(el peso real de las antocianinas CLM recolectadas) × 100.

^bLos datos con el mismo superíndice en la misma fila no son significativamente diferentes (P> 0.05).

Table 4. Antioxidant activities (DPPH, ABTS and FRAP), α-glucosidase inhibition activity (IC₅₀) and fluorescence quenching constant (K_FQ) of CLM anthocyanins from the samples before and after in vitro gastrointestinal digestion.^a,b

Tabla 4. Actividades antioxidantes (DPPH, ABTS y FRAP), actividad de inhibición de la α-glucosidasa (IC₅₀) y constante de desactivación de la fluorescencia (K_FQ) de las antocianinas CLM de las muestras tomadas antes y después de la digestión gastrointestinal in vitro.^a,b

	DPPH	ABTS	FRAP	IC50 (×10^−4 mg/mL)	KFQ (mL/mg)
Hedong
Before	45.11 ± 1.03^b	104.75 ± 2.39^b	95.89 ± 1.32^b	13.2 ± 0.2^a	2.3630 ± 0.02^a
After	65.13 ± 1.41^a	126.17 ± 3.10^a	115.79 ± 1.63^a	2.0 ± 0.1^b	2.3642 ± 0.01^a
Delingha
Before	37.85 ± 0.99^b	96.51 ± 1.80^b	84.36 ± 1.01^b	15.7 ± 0.1^a	1.7146 ± 0.02^a
After	49.50 ± 1.54^a	114.50 ± 2.86^a	103.10 ± 1.06^a	4.8 ± 0.2^b	1.7124 ± 0.02^a
Nomuhong
Before	42.67 ± 0.85^b	102.07 ± 2.00^b	91.77 ± 0.94^b	14.3 ± 0.2^a	1.9646 ± 0.01^a
After	63.88 ± 1.22^a	122.83 ± 2.19^a	112.62 ± 2.55^a	3.1 ± 0.2^b	1.9782 ± 0.02^a

^a Data with the same superscript in the same column are not significantly different (P> 0.05).

^b DPPH, ABTS and FRAP content were expressed as μmol TE/g DW.

^aLos datos con el mismo superíndice en la misma columna no son significativamente diferentes (P> 0.05).

^bEl contenido de DPPH, ABTS y FRAP se expresó como μmol TE/g DW.

Figure 4. Fluorescence spectra of α-glucosidase in the absence (black line) and presence (colored lines) of CLM anthocyanins from the samples before (a) and after (b) in vitro gastrointestinal digestion. From top to bottom, the concentrations of CLM anthocyanins are 0.0, 0.05, 0.1, 0.2, 0.4 and 0.8 mg/mL; Stern-Volmer plots for fluorescence quenching of α-glucosidase by CLM anthocyanins before (c) and after (d) in vitro gastrointestinal digestion. The equations for them were fitted according to Eq. (4).

Figura 4. Espectros de fluorescencia de α-glucosidasa en ausencia (línea negra) y presencia (líneas coloreadas) de antocianinas CLM de las muestras antes (a) y después (b) de la digestión gastrointestinal in vitro. De arriba a abajo, las concentraciones de antocianinas CLM son 0.0, 0.05, 0.1, 0.2, 0.4 y 0.8 mg/mL; diagramas de Stern-Volmer para la desactivación de la fluorescencia de la α-glucosidasa por las antocianinas CLM antes (c) y después (d) de la digestión gastrointestinal in vitro. Las ecuaciones para ello se ajustaron de acuerdo con la ecuación (4).