Developed validation for simultaneous determination of three Di-caffeoylquinic acid derivatives from the Leaf of <i>Eribotrya japonica</i> Lindl. by HPLC-DAD

SEON, Ho-Young; KIM, Hyun-Hee; YIM, Soon-Ho

doi:10.1590/fst.68121

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Original Article • Food Sci. Technol (Campinas) 42 • 2022 • https://doi.org/10.1590/fst.68121 copy

Developed validation for simultaneous determination of three Di-caffeoylquinic acid derivatives from the Leaf of Eribotrya japonica Lindl. by HPLC-DAD

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Abstract

The aim of this study was to developed HPLC simultaneous analysis method for the determination of Caffeoylquinic acid isolated from the leaf of Eribotrya japonica Lindl. extract. The phenolic compounds in E. japonica leaves were isolated and identified by HPLC, NMR, Mass etc. The quantitative analysis was carried out HPLC-DAD using C18 column with gradient elution of water and acetonitrile at 280 nM for 60 min. The method was successfully validated by specificity, LOD (linearity, limit of detection), LOQ (limit of quantification), precision and recovery. The three phenolic compounds were isolated from E. japonica leaves including 5-Caffeoylquinic acid (5-CQA), 4-Caffeoylquinic acid (4-CQA) and 3-Caffeoylquinic acid (3-CQA). In a quantitative analysis of three phenolic compounds were 3-CQA 7.82-14.48 µg/mL, 4-CQA 3.75-12.58 µg/mL and 5-CQA 3.60-7.82 µg/mL using the standard chemicals, respectively. The developed HPLC validation method would be applicable for simultaneous quantitative analysis of phenolic compounds in E. japonica leaves extracts.

Keywords:
validation; simultaneous analysis; 5-Caffeoylquinic acid; 3-Caffeoylquinic acid; 4-Caffeoylquinic acid

Compounds	Retention time	Regression equation	R²	LOD (µg/mL)	LOQ (µg/mL)
5-CQA (1)	19.227	y = 16.629x+4.275	0.9999	0.245	0.741
3-CQA (2)	29.776	y = 19.463x+11.4613	0.9996	0.334	1.043
4-CQA (3)	31.562	y = 18.531x+4.439	0.9999	0.112	0.340

Compound	Concentration	Intra-day¹ 1) Intra-day: three time per day; )		Inter-day² 2) Inter- day: one time analysis of compounds 1-3 per day for 3 days; )
Compound	Concentration	Mean ± SD³ 3) Values are mean±standard deviation in triplicate (n = 6); ) (µg/mL)	RSD⁴ 4) Relative standard deviation. ) (%)	Mean ± SD (µg/mL)	RSD (%)
5-CQA (1)	9.54	9.53 ± 0.05	0.52	9.51 ± 0.00	0.00
	38.15	38.27 ± 0.14	0.36	37.70 ± 0.10	0.26
	76.30	76.15 ± 0.34	0.44	75.13 ± 0.24	0.31
3-CQA (2)	9.54	9.15 ± 0.03	0.32	9.20 ± 0.08	0.86
	38.15	38.96 ± 0.06	0.15	38.23 ± 0.11	0.28
	76.30	75.91 ± 0.28	0.36	75.24 ± 0.12	0.15
4-CQA (3)	9.54	9.40 ± 0.01	0.10	9.37 ± 0.02	0.21
	38.15	38.47 ± 0.03	0.07	39.77 ± 0.02	0.05
	76.30	76.51 ± 0.15	0.19	75.04 ± 0.04	0.05

Compound	Spiked amount (µg/mL)	Measured amount (µg/mL)	RSD(%)	Recovery (%)
5-CQA	9.54	9.59 ± 0.05	0.52	99.9
				100.9
				100.8
	38.15	38.27 ± 0.14	0.36	99.9
				100.6
				100.5
	76.30	76.15 ± 0.34	0.44	99.6
				99.5
				100.3
3-CQA	9.54	9.15 ± 0.03	0.32	95.6
				96.1
				96.1
	38.15	38.96 ± 0.06	0.15	102.0
				102.3
				102.1
	76.30	75.91 ± 0.28	0.36	99.6
				99.1
				99.8
4-CQA	9.54	9.40 ± 0.01	0.10	98.3
				98.6
				98.6
	38.15	38.47 ± 0.03	0.07	100.8
				100.8
				100.9
	76.30	76.51 ± 0.15	0.19	100.3
				100.1
				100.5

Compound		Content (µg/mL of extract)	RSD (%)
Hot Water	5-CQA	7.82 ± 0.02	0.23
	3-CQA	12.20 ± 0.06	0.52
	4-CQA	12.58 ± 0.11	0.87
20% E	5-CQA	3.60 ± 0.02	0.61
	3-CQA	9.28 ± 0.01	0.08
	4-CQA	4.27 ± 0.04	0.98
40% E	5-CQA	4.68 ± 0.04	0.75
	3-CQA	14.48 ± 0.06	0.42
	4-CQA	7.02 ± 0.09	1.24
60% E	5-CQA	4.29 ± 0.05	1.13
	3-CQA	10.60 ± 0.00	0.03
	4-CQA	4.78 ± 0.03	0.55
80% E	5-CQA	4.18 ± 0.03	0.74
	3-CQA	10.00 ± 0.01	0.14
	4-CQA	4.39 ± 0.01	026
100% E	5-CQA	4.54 ± 0.04	0.97
	3-CQA	7.82 ± 0.02	0.23
	4-CQA	3.74 ± 0.02	0.40

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[1] *Corresponding author: virshyim@dsu.ac.kr