Anticholinergic and antioxidant activities of avocado (Folium perseae) leaves – phytochemical content by LC-MS/MS analysis

Figures & data

Table 1. LC-MS/MS parameter of selected compounds in water and ethanol extracts of avocado (Folium perseae) leaves [EEFP: ethanol extract of avocado (F. perseae) leaves, WEFP: water extract of avocado (F. perseae) leaves].

	Compounds	Parent ion	Daughter ion		Collision energy (V)
1	Gallic acid	168.6		124	13
2	Herniarin	177		121	12
3	Kaempferol	287		152.3	30
4	Quercitrin	471.9		309.9	16
5	Fumaric acid	115		71	8
6	Pyrogallol	125		80	16
7	Caffeic acid	179		135	10
8	Quercetin	301		178.5	16
9	Ellagic acid	301		228.3	25
10	Chlorogenic acid	353		191	14
11	Rosmarinic acid	359.2		160.5	15
12	Luteolin-7-glucoside	447		284.5	14
13	Luteolin-5-glucoside	447		289.5	20
14	Kaempferon-3-O-rutinoside	593		284.4	18
15	Rutin	609		301	16
16	Curcumin*	369.3		176.9	20

Notes: * Used as internal standard.

Table 2. Validation and uncertainty parameters of selected compounds.

Compounds	Linear regression equation	r^2	LOD (mg/L)	LOQ (mg/L)	RSD (%)
Gallic acid	y = 0.0569x + 0.0177	0.991	0.002	0.008	4.85
Herniarin	y = 0.0325x + 0.0107	0.9917	0.370	1.233	9.47
Kaempferol	y = 0.0230x + 0.0116	0.984	0.002	0.008	5.47
Quercitrin	y = 0.0290x + 0.0058	0.9918	0.001	0.002	4.28
Fumaric acid	y = 0.0569x + 0.0177	0.991	0.003	0.01	5.44
Pyrogallol	y = 0.0438x + 0.0073	0.980	0.001	0.002	5.47
Caffeic acid	y = 0.3300x + 0.0036	0.992	0.028	0.093	8.04
Quercetin	y = 0.1150x + 0.0078	0.993	0.001	0.002	0.11
Ellagic acid	y = 0.2358x + 0.005	0.9950	0.020	0.068	0.11
Chlorogenic acid	y = 0.2620x + 0.0674	0.998	0.445	1.483	5.45
Rosmarinic acid	y = 0.1960x + 0.0043	0.998	0.022	0.072	3.73
Luteolin-7-glucoside	y = 0.1350x + 0.0246	0.995	0.022	0.072	8.56
Luteolin-5-glucoside	y = 0.2300x + 0.0413	0.992	0.01	0.034	1.12
Kaempferon-3-O-rutinoside	y = 0.1080x + 0.0135	0.997	0.014	0.045	8.15
Rutin	y = 0.0232x + 0.0008	0.996	0.01	0.034	7.90

Figure 2. (a) Fe³⁺-Fe²⁺ reductive potential of different concentrations (10–30 μg/mL) of avocado leaf extracts and reference antioxidants. (b) Cu²⁺ reducing ability of different concentrations (10–30 μg/mL) of avocado leaf extracts and reference antioxidants. (c) Fe³⁺-TPTZ-Fe²⁺-TPTZ reducing ability of different concentrations (10–30 μg/mL) of avocado leaf extracts and reference antioxidants [EEFP: ethanol extract of avocado (Folium perseae) leaves, WEFP: water extract of avocado (F. perseae) leaves, BHA, butylated hydroxyanisole, BHT, butylated hydroxytoluene].

Table 3. Determination of reducing power by potassium ferricyanide reduction method and, CUPRAC and FRAP methods of avocado (Folium perseae) leaves at the same concentration (30 μg/mL) [EEFP: ethanol extract of avocado (F. perseae) leaves, WEFP: water extract of avocado (F. perseae) leaves].

Antioxidants	Fe^3+-Fe^2+ reducing		Cu^+-Cu^+ reducing		Fe^3+-TPTZ reducing
	λ700	r^2	λ450	r^2	λ593	r^2
BHA	2.170 ± 0.005	0.9616	2.396 ± 0.018	0.9107	2.853 ± 0.003	0.8282
BHT	1.490 ± 0.002	0.9950	2.020 ± 0.004	0.9206	2.026 ± 0.002	0.8870
Trolox	1.170 ± 0.001	0.9955	1.452 ± 0.050	0.9970	2.102 ± 0.003	0.9201
α-Tocopherol	1.101 ± 0.006	0.9631	1.262 ± 0.018	0.9920	1.855 ± 0.001	0.9175
EEFP	0.606 ± 0.002	0.9691	0.690 ± 0.001	0.9592	1.316 ± 0.001	0.8102
WEFP	0.401 ± 0.002	0.9538	0.309 ± 0.002	0.9926	0.982 ± 0.002	0.8945

Figure 3. (a) DPPH free radical scavenging activity of different concentrations (10–30 μg/mL) of avocado leaf (Folium perseae) extracts and reference antioxidants. (b) ABTS radical scavenging activity of different concentrations (10–30 μg/mL) of avocado leaf extracts and standard antioxidant compounds [EEFP: ethanol extract of avocado (F. perseae) leaves, WEFP: water extract of avocado (F. perseae) leaves, BHA, butylated hydroxyanisole, BHT, butylated hydroxytoluene].

Table 4. Determination of half maximal scavenging concentrations (IC₅₀, μg/mL) of DPPH· and ABTS^•+ scavenging, and AChE/BChE inhibition assays for EEFP and WEFP [EEFP: ethanol extract of avocado (F. perseae) leaves, WEFP: water extract of avocado (F. perseae) leaves].

Antioxidants	DPPH^. scavenging		ABTS^•+ scavenging		AChE inhibition		BChE inhibition
	IC50	r^2	IC50	r^2	IC50	r^2	IC50	r^2
BHA	45.6456	0.9482	18.8416	0.7539	–	–	–	–
BHT	87.9514	0.8916	16.9169	0.9159	–	–	–	–
α-Tocopherol	85.8573	0.9781	125.8622	0.9217	–	–	–	–
Trolox	58.1614	0.9585	17.0084	0.7263	–	–	–	–
EEFP	240.400	0.9989	286.0504	0.9829	0.0168	0.9803	0.0214	0.9912
WEFP	601.001	0.9876	524.4258	0.9883	0.0171	0.9911	0.0226	0.9945
Tacrine	–	–	–	–	1.0376	0.9922	0.0870	0.9862

Figure 1. Chromatogram of secondary metabolites of avocado leaf (F. perseae) in EEFP [EEFP: ethanol extract of avocado (F. perseae) leaves].

Table 5. The quantity (mg/kg) of secondary metabolites in avocado (F. perseae) leaves extracts [EEFP: ethanol extract of avocado (F. perseae) leaves, WEFP: water extract of avocado (F. perseae) leaves].

Compounds	EEFP	WEFP
Gallic acid	5.41 ± 0.38	–
Herniarin	17.98 ± 1.81	0.60 ± 0.06
Kaempferol	663.54 ± 46.83	50.15 ± 3.54
Quercitrin	58.61 ± 3.74	15.10 ± 0.96
Fumaric acid	214.32 ± 14.86	59.18 ± 4.10
Pyrogallol	–	122.25 ± 8.14
Caffeic acid	32.74 ± 6.48	10.83 ± 2.14
Quercetin-3-O-arabinoside	253.18 ± 33.66	–
Quercetin	16.20 ± 2.15	–
Ellagic acid	5.56 ± 0.37	–
Chlorogenic acid	852.81 ± 118.09	28.83 ± 3.99
Rosmarinic acid	5.46 ± 0.42	–
Luteolin-7-glucoside	43.15 ± 4.39	1.08 ± 0.11
Luteolin-5-glucoside	18.44 ± 1.19	–
Kaempferon-3-O-rutinoside	25.45 ± 2.30	9.45 ± 0.85
Rutin	68.41 ± 4.48	26.05 ± 1.71
Isorhamnetin	34.37 ± 9.33	1.84 ± 0.12