Quantification of chlorogenic acid, rosmarinic acid, and caffeic acid contents in selected Thai medicinal plants using RP-HPLC-DAD

Chaowuttikul, Chayanon; Palanuvej, Chanida; Ruangrungsi, Nijsiri

doi:10.1590/s2175-97902019000317547

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Brazilian Journal of Pharmaceutical Sciences

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Article • Braz. J. Pharm. Sci. 56 • 2020 • https://doi.org/10.1590/s2175-97902019000317547 copy

Quantification of chlorogenic acid, rosmarinic acid, and caffeic acid contents in selected Thai medicinal plants using RP-HPLC-DAD

Authorship SCIMAGO INSTITUTIONS RANKINGS

The chlorogenic acid, rosmarinic acid, and caffeic acid contents in 100 selected plants were determined using reversed phase high performance liquid chromatography equipped with diode array detector. The optimum condition was 0.2% phosphoric acid in water (solvent A) and methanol (solvent B) as the mobile phase, which was set at 45% B for 20 minutes at a flow rate of 1.2 mL/min. The column temperature was maintained at 30 ºC and the detection wavelength was 325 nm. Among 100 selected plants, 39.64% contained all 3 compounds, 40.54% contained 2 compounds, 14.41% contained only 1 compound, and 5.41% could not detect any of the 3 compounds. The highest contents of chlorogenic acid, rosmarinic acid, and caffeic acid were found in Lonicera japonica flowering buds, Melissa officinalis leaves, and Coffea canephora seeds at the concentration of 9.900 ± 0.004, 19.908 ± 0.171, and 1.233 ± 0.003 g/100 g of dried plant, respectively.

Keywords:
Chlorogenic acid; Rosmarinic acid; Caffeic acid; RP-HPLC-DAD; Medicinal plant

Voucher specimen Number	Scientific plant name	Plant parts used	% yield (g/100 g)	Content (g/100 g of dried plant)
Voucher specimen Number	Scientific plant name	Plant parts used	% yield (g/100 g)	Chlorogenic acid	Rosmarinic acid	Caffeic acid
	Family: Acanthaceae
CCPh001	Andrographis paniculata (Burm.f.) Wall. ex Nees	Leaves	25.595	0.625 ± 0.001	0.338 ± 0.000	0.045 ± 0.000
	Family: Alliaceae
CCPh002	Allium sativum L.	Bulbs	10.393	-	-	0.005 ± 0.000
CCPh003	Allium cepa L.	Bulbs	66.467	0.034 ± 0.000	-	-
	Family: Amaranthaceae
CCPh004	Spinacia oleracea L.	Leaves	21.103	-	0.039 ± 0.000	0.037 ± 0.000
	Family: Anacardiaceae
CCPh005	Mangifera indica L. cv. Okrong	Leaves	28.698	0.354 ± 0.003	-	1.008 ± 0.008
	Family: Apiaceae
CCPh006	Anethum graveolens L.	Aerial part	28.714	7.361 ± 0.038	-	0.184 ± 0.002
CCPh007	Apium graveolens L.	Aerial part	36.739	0.913 ± 0.003	2.880 ± 0.018	0.088 ± 0.001
CCPh008	Apium graveolens L. var. secalinum	Aerial part	36.066	2.574 ± 0.011	2.556 ± 0.014	0.112 ± 0.000
CCPh009	Centella asiatica (L.) Urb.	Aerial part	31.917	0.848 ± 0.003	0.910 ± 0.003	0.086 ± 0.002
CCPh010	Coriandrum sativum L.	Seeds	4.770	0.028 ± 0.000	0.061 ± 0.001	0.024 ± 0.000
CCPh011	Daucus carota L. subsp. sativus (Hoffm.) Arcang.	Roots	52.063	0.326 ± 0.003	-	0.085 ± 0.000
CCPh012	Eryngium foetidum L.	Leaves	30.451	4.979 ± 0.006	4.302 ± 0.100	0.160 ± 0.007
CCPh013	Petroselinum crispum (Mill.) Nyman ex A.W. Hill	Aerial part	31.826	0.058 ± 0.004	9.153 ± 0.602	0.048 ± 0.001
	Family: Apocynaceae
CCPh014	Telosma cordata (Burm. f.) Merr.	Flowers	34.584	0.350 ± 0.000	-	0.184 ± 0.002
	Family: Asteraceae
CCPh015	Artemisia dracunculus L.	Aerial part	28.885	5.253 ± 0.047	5.359 ± 0.037	0.062 ± 0.008
CCPh016	Artemisia pallens Wall. ex DC.	Aerial part	8.141	0.042 ± 0.000	-	0.011 ± 0.000
CCPh017	Chromolaena odorata (L.) R. M. King & H. Rob.	Leaves	28.239	4.137 ± 0.035	0.279 ± 0.009	0.621 ± 0.018
CCPh018	Gnaphalium polycaulon Pers.	Aerial part	23.409	0.699 ± 0.009	1.207 ± 0.012	0.236 ± 0.009
CCPh019	Helianthus annuus L.	Pericarps	2.880	0.038 ± 0.000	0.004 ± 0.000	0.003 ± 0.000
CCPh020	Helianthus annuus L.	Seeds	11.523	2.144 ± 0.006	0.008 ± 0.000	0.110 ± 0.000
CCPh021	Helianthus annuus L.	Sprouts	30.726	2.199 ± 0.005	3.117 ± 0.029	0.093 ± 0.002
CCPh022	Lactuca sativa L.	Leaves	24.088	1.396 ± 0.036	0.343 ± 0.001	0.258 ± 0.002
	Family: Brassicaceae
CCPh023	Brassica juncea (L.) Czern.	Leaves	28.414	0.351 ± 0.002	-	0.083 ± 0.001
CCPh024	Brassica oleracea L. Group Capitata	Aerial part	38.510	0.172 ± 0.002	-	-
CCPh025	Brassica rapa L. Group Pekinensis	Aerial part	41.573	0.154 ± 0.001	-	-
CCPh026	Raphanus sativus L.	Roots	52.566	0.072 ± 0.002	-	0.033 ± 0.001
	Family: Caprifoliaceae
CCPh027	Lonicera japonica Thunb.	Flowering bud	32.734	9.896 ± 0.004	2.543 ± 0.007	0.195 ± 0.002
	Family: Caricaceae
CCPh028	Carica papaya L.	Leaves	17.759	0.049 ± 0.000	0.023 ± 0.001	0.508 ± 0.000
	Family: Convolvulaceae
CCPh029	Ipomoea aquatica Forssk.	Aerial part	30.503	2.541 ± 0.014	8.303 ± 0.042	0.113 ± 0.002
	Family: Cucurbitaceae
CCPh030	Momordica charantia L. (Thai varieties)	Fruits	28.359	0.042 ± 0.000	-	<LOQ
CCPh031	Momordica charantia L. (Chinese varieties)	Fruits	35.728	0.085 ± 0.001	-	<LOQ
	Family: Eucommiaceae
CCPh032	Eucommia ulmoides Oliv.	Stem barks	10.441	0.039 ± 0.000	-	0.019 ± 0.000
	Family: Euphorbiaceae
CCPh033	Euphorbia hirta L.	Aerial part	11.150	0.234 ± 0.000	-	0.021 ± 0.000
CCPh034	Phyllanthus emblica L.	Fruits	28.671	0.300 ± 0.001	0.077 ± 0.000	0.029 ± 0.000
CCPh035	Ricinus communis L.	Leaves	22.599	0.039 ± 0.000	-	0.352 ± 0.004
	Family: Fabaceae
CCPh036	Pisum sativum L.	Fruits	45.838	-	-	-
CCPh037	Pisum sativum L. var. macrocarpon	Fruits	43.713	-	-	-
CCPh038	Sesbania grandiflora (L.) Poir.	Flowers	42.093	0.016 ± 0.000	0.204 ± 0.007	<LOQ
CCPh039	Sesbania grandiflora (L.) Poir.	Stem barks	3.960	0.009 ± 0.000	-	0.006 ± 0.000
	Family: Gnetaceae
CCPh040	Gnetum gnemon L. var. tenerum Markgr.	Leaves	22.424	0.092 ± 0.001	-	-
	Family: Labiatae
CCPh041	Hyptis suaveolens (L.) Poit.	Aerial part	11.194	0.078 ± 0.001	3.961 ± 0.006	0.111 ± 0.000
CCPh042	Leonotis nepetifolia (L.) R. Br.	Leaves	22.287	0.171 ± 0.001	-	0.418 ± 0.010
CCPh043	Leonurus sibiricus L.	Aerial part	14.098	0.045 ± 0.003	-	0.012 ± 0.000
CCPh044	Melissa officinalis L.	Leaves	20.371	0.048 ± 0.004	19.908 ± 0.171	0.174 ± 0.006
CCPh045	Mentha arvensis L. var. piperascens Malinv.	Leaves	19.968	1.038 ± 0.002	6.809 ± 0.086	0.108 ± 0.001
CCPh046	Mentha cordifolia Opiz ex Fresen	Leaves	21.461	0.117 ± 0.000	7.537 ± 0.010	0.100 ± 0.001
CCPh047	Ocimum africanum Lour.	Leaves	15.466	0.033 ± 0.000	1.691 ± 0.002	0.092 ± 0.000
CCPh048	Ocimum basilicum L.	Leaves	15.756	0.243 ± 0.001	0.597 ± 0.003	0.311 ± 0.001
CCPh049	Ocimum gratissimum L. var. macrophyllum Briq.	Leaves	19.372	0.189 ± 0.002	1.756 ± 0.001	0.240 ± 0.003
CCPh050	Ocimum tenuiflorum L.	Leaves	17.317	0.215 ± 0.001	2.292 ± 0.003	0.112 ± 0.002
CCPh051	Origanum majorana L.	Leaves	24.119	-	7.954 ± 0.028	0.065 ± 0.003
CCPh052	Origanum vulgare L.	Leaves	31.593	0.123 ± 0.005	9.902 ± 0.091	0.410 ± 0.009
CCPh053	Orthosiphon aristatus (Blume) Miq.	Leaves	11.332	0.072 ± 0.002	2.101 ± 0.003	0.115 ± 0.001
CCPh054	Perilla frutescens (L.) Britton	Leaves	21.621	0.042 ± 0.001	13.185 ± 0.021	0.189 ± 0.001
	Family: Labiatae
CCPh055	Plectranthus amboinicus (Lour.) Spreng.	Leaves	12.369	0.049 ± 0.001	0.279 ± 0.003	0.121 ± 0.001
CCPh056	Plectranthus rotundifolius (Poir.) Spreng.	Leaves	22.849	-	0.334 ± 0.003	0.086 ± 0.001
CCPh057	Plectranthus rotundifolius (Poir.) Spreng.	Tubers	6.628	-	0.669 ± 0.005	0.270 ± 0.004
CCPh058	Plectranthus scutellarioides (L.) R. Br.	Leaves	26.274	-	2.594 ±0.016	0.217 ± 0.001
CCPh059	Rosmarinus officinalis L.	Aerial part	17.989	0.124 ± 0.005	2.611 ± 0.023	0.146 ± 0.010
CCPh060	Salvia hispanica L.	Seeds	5.427	0.029 ± 0.000	0.576 ± 0.025	0.007 ± 0.000
CCPh061	Salvia officinalis L.	Aerial part	20.719	-	6.829 ± 0.070	0.166 ± 0.009
CCPh062	Thymus citriodorus (Pers.) Schreb.	Aerial part	19.963	0.165 ± 0.001	10.176 ± 0.417	0.128 ± 0.010
CCPh063	Thymus vulgaris L.	Aerial part	21.499	0.093 ± 0.005	4.349 ± 0.025	0.155 ± 0.002
	Family: Lauraceae
CCPh064	Persea americana Mill.	Flesh	16.142	0.029 ± 0.000	-	-
CCPh065	Persea americana Mill.	Peels	15.259	0.539 ± 0.001	-	0.008 ± 0.000
CCPh066	Persea americana Mill.	Seeds	22.143	1.381 ± 0.005	-	-
	Family: Malvaceae
CCPh067	Hibiscus sabdariffa L.	Leaves	31.829	1.117 ± 0.002	-	0.351 ± 0.002
	Family: Meliaceae
CCPh068	Azadirachta indica A. Juss.	Leaves	16.665	-	-	-
	Family: Moraceae
CCPh069	Morus alba L.	Leaves	20.045	3.028 ± 0.005	0.050 ± 0.001	0.025 ± 0.001
	Family: Moringaceae
CCPh070	Moringa oleifera Lam.	Leaves	16.509	0.525 ± 0.002	-	0.030 ± 0.000
CCPh071	Moringa oleifera Lam.	Seeds	8.414	-	-	0.004 ± 0.000
	Family: Myrtaceae
CCPh072	Psidium guajava L.	Fruits	50.035	-	-	0.022 ± 0.000
CCPh073	Syzygium antisepticum (Blume) Merr. & L. M. Perry	Leaves	23.821	-	-	-
	Family: Oxalidaceae
CCPh074	Averrhoa carambola L.	Fruits	63.412	0.045 ± 0.001	-	0.009 ± 0.000
	Family: Piperaceae
CCPh075	Piper betle L.	Leaves	21.431	0.136 ± 0.000	-	0.041 ± 0.000
CCPh076	Piper nigrum L. (Black pepper)	Fruits	7.985	-	-	-
CCPh077	Piper nigrum L. (White pepper)	Seeds	5.619	-	-	-
	Family: Poaceae
CCPh078	Cymbopogon citratus (DC.) Stapf	Rhizomes	26.886	0.132 ± 0.002	-	0.073 ± 0.003
	Family: Polygonaceae
CCPh079	Persicaria odorata (Lour.) Soják	Leaves	17.391	0.289 ± 0.017	-	0.091 ± 0.000
	Family: Punicaceae
CCPh080	Punica granatum L. var. granatum	Leaves	43.525	0.194 ± 0.009	-	0.305 ± 0.006
CCPh081	Punica granatum L. var. granatum	Peels	40.172	-	0.049 ± 0.000	0.363 ± 0.003
	Family: Rosaceae
CCPh082	Fragaria vesca L.	Fruits	70.506	0.046 ± 0.001	-	-
CCPh083	Malus domestica Borkh.	Fruits	85.332	0.277 ± 0.001	-	-
CCPh084	Pyrus communis L.	Fruits	74.522	0.407 ± 0.001	-	-
	Family: Rubiaceae
CCPh085	Coffea arabica L.	Seeds	8.228	5.967 ± 0.007	0.845 ± 0.002	0.380 ± 0.001
CCPh086	Coffea canephora Pierre ex A. Froehner	Seeds	12.262	7.843 ± 0.037	1.663 ± 0.007	1.233 ± 0.003
CCPh087	Morinda citrifolia L.	Fruits	23.686	0.046 ± 0.001	-	0.010 ± 0.000
CCPh088	Morinda citrifolia L.	Leaves	28.441	0.081 ± 0.000	-	-
	Family: Scrophulariaceae
CCPh089	Limnophila aromatica (Lam.) Merr.	Aerial part	16.237	0.649 ± 0.001	-	0.058 ± 0.001
	Family: Solanaceae
CCPh090	Capsicum annuum L. (Green bell pepper)	Fruits	51.863	0.148 ± 0.000	-	0.025 ± 0.000
CCPh091	Capsicum annuum L. (Orange bell pepper)	Fruits	55.366	0.169 ± 0.001	-	0.026 ± 0.001
CCPh092	Capsicum annuum L. (Red bell pepper)	Fruits	58.196	0.164 ± 0.002	-	0.085 ± 0.002
CCPh093	Capsicum annuum L. (Yellow bell pepper)	Fruits	58.237	0.175 ± 0.003	-	0.026 ± 0.001
CCPh094	Nicotiana tabacum L.	Leaves	30.838	3.317 ± 0.001	-	0.057 ± 0.000
CCPh095	Physalis angulata L.	Aerial part	12.733	0.196 ± 0.002	-	0.012 ± 0.000
CCPh096	Physalis peruviana L.	Fruits	61.376	0.097 ± 0.001	-	<LOQ
CCPh097	Physalis peruviana L.	Calyx	13.857	0.474 ± 0.001	-	0.086 ± 0.001
CCPh098	Solanum lycopersicum L. var. cerasiforme	Fruits	45.374	0.225 ± 0.010	-	0.037 ± 0.000
CCPh099	Solanum lycopersicum L. var. lycopersicum	Fruits	60.356	0.280 ± 0.009	-	0.035 ± 0.000
	Family: Strychnaceae
CCPh100	Strychnos nux-vomica L.	Seeds	3.228	0.389 ± 0.000	-	0.004 ± 0.000
	Family: Theaceae
CCPh 101	Camellia sinensis (L.) Kuntze var. assamica (Mast.) Kitam.	Leaves	46.363	0.719 ± 0.003	-	0.051 ± 0.001
	Family: Thunbergiaceae
CCPh 102	Thunbergia laurifolia Lindl.	Leaves	14.292	0.082 ± 0.004	11.487 ± 0.019	0.218 ± 0.001
	Family: Verbenaceae
CCPh 103	Clerodendrum calamitosum L.	Leaves	20.401	0.023 ± 0.001	-	0.010 ± 0.000
CCPh 104	Clerodendrum indicum (L.) Kuntze	Leaves	24.400	-	-	0.011 ± 0.001
CCPh 105	Clerodendrum quadriloculare (Blanco) Merr.	Leaves	21.407	0.066 ± 0.001	-	-
CCPh 106	Clerodendrum serratum (L.) Moon	Leaves	33.686	1.804 ± 0.006	-	-
CCPh 107	Clerodendrum thomsoniae Balf. f.	Leaves	19.390	0.018 ± 0.002	0.903 ± 0.004	0.077 ± 0.002
CCPh 108	Vitex agnus-castus L.	Leaves	32.946	5.557 ± 0.068	3.083 ± 0.037	0.084 ± 0.001
CCPh 109	Vitex negundo L.	Leaves	28.441	1.238 ± 0.002	1.135 ± 0.005	0.054 ± 0.004
CCPh 110	Vitex trifolia L. subsp. litoralis Steenis	Leaves	28.545	0.379 ± 0.003	1.497 ± 0.008	0.167 ± 0.003
CCPh 111	Vitex trifolia L. subsp. trifolia	Leaves	31.403	2.180 ± 0.056	2.745 ± 0.098	0.177 ± 0.006

Parameter	Result	Acceptance criteria
Retention factor (k)	4.30 ± 0.01	k ˃ 2
Theoretical plate number (N)	2745.17 ± 158.17	N ˃ 2000
Tailing factor (T_f)	1.027 ± 0.07	T_f ≤ 2

Compounds	Spike concentration (µg/mL)	% recovery (n = 3)	%RSD
Compounds	Spike concentration (µg/mL)	% recovery (n = 3)	Repeatability precision (n = 3)	Intermediate precision (n = 5)
Chlorogenic acid	10	108.632	0.130	0.989
	25	103.976	0.077	0.699
	50	107.396	0.054	1.770
Rosmarinic acid	10	97.230	0.259	1.522
	25	99.089	0.234	1.039
	50	98.328	0.135	1.415
Caffeic acid	10	100.447	0.169	6.468
	25	99.407	0.046	5.795
	50	100.851	0.074	3.119

Compounds	% RSD of sample
	Flow rate		Temperature		Wavelength
	Rt	Area	Rt	Area	Rt	Area
Chlorogenic acid	0.31	0.50	0.79	0.78	0.06	1.14
Rosmarinic acid	0.19	0.66	2.63	0.89	0.02	1.33
Caffeic acid	0.27	0.95	1.11	3.14	0.07	2.08

Compounds	% RSD of standard compounds
	Flow rate		Temperature		Wavelength
	Rt	Area	Rt	Area	Rt	Area
Chlorogenic acid	0.24	0.33	0.78	0.14	0.14	1.13
Rosmarinic acid	0.11	1.38	2.57	1.10	0.03	2.41
Caffeic acid	0.23	0.29	1.20	0.25	0.09	0.87

Universidade de São Paulo, Faculdade de Ciências Farmacêuticas Av. Prof. Lineu Prestes, n. 580, 05508-000 S. Paulo/SP Brasil, Tel.: (55 11) 3091-3824 - São Paulo - SP - Brazil
E-mail: bjps@usp.br

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[1] *Correspondence: C. Palanuvej. College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand. E-mail: chanida.p@chula.ac.th.