An optimization approach of dynamic maceration of <i>Centella asiatica</i> to obtain the highest content of four centelloids by response surface methodology

Monton, Chaowalit; Settharaksa, Sukanya; Luprasong, Chitradee; Songsak, Thanapat

doi:10.1016/j.bjp.2019.01.001

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Original articles • Rev. bras. farmacogn. 29 (2) • Mar-Apr 2019 • https://doi.org/10.1016/j.bjp.2019.01.001 copy

An optimization approach of dynamic maceration of Centella asiatica to obtain the highest content of four centelloids by response surface methodology

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

Centella asiatica (L.) Urb., Apiaceae, is commonly used as food, food supplement, and medicine. Development of the extraction process to obtain the high extent of the active compound is necessary. So, the response surface methodology was used in this work to optimize the dynamic maceration of C. asiatica to obtain the highest content of the four centelloids including asiatic acid, madecassic acid, asiaticoside, and madecassoside. Two factors: extraction temperature and extraction time, were studied. The content of four centelloids was observed. After the extraction of C. asiatica using ethanol, the content of four centelloids was analyzed using validated high performance liquid chromatography. The optimization result showed that madecassoside and asiaticoside had a similar pattern of the contour plots and response surfaces. These two centelloids were highly extracted at a high extraction time and high extraction temperature. The other two centelloids had the same pattern, they had a high content at a high temperature and time as well as at a low temperature and time. The simultaneous highest content of four centelloids was achieved when extracted at 60 °C for 120 min. The optimal condition could be used as standard condition for extraction of C. asiatica to provide the highest content of four centelloids.

Keywords:
Centelloids; Dynamic maceration; HPLC analysis; Optimization; Response surface methodology

Factor	Condition
	1	2	3	4	5	6	7	8	9	10
X ₁ ^a a X1 was extraction temperature; -√2 = 40 °C, -1 = 42.9 °C, 0 = 50 °C, 1 = 57.1 °C, and √2 = 60 °C.	-1	1	-1	1	-√2	√2	0	0	0	0
X ₂ ^b b X2 was extraction time; -√2 = 60 min, -1 = 68.8 min, 0 = 90 min, 1 = 111.2 min, and √2 = 120 min.	-1	-1	1	1	0	0	-√2	√2	0	0

Compounds	Equation	R ²	Range (µg/ml)	DL (µg/ml)	QL (µg/ml)
Madecassoside	$y = 3922.5 x + 1354.7$	0.9999	0.78-50	0.21	0.61
Asiaticoside	$y = 5199.6 x + 1398.3$	0.9999	0.78-50	0.20	0.60
Madecassic acid	$y = 16018 x + 2342.6$	0.9999	0.78-50	0.19	0.59
Asiatic acid	$y = 22659 x + 5060.4$	0.9998	0.78-50	0.22	0.65

Compounds	Conc. (µg/ml)	Precision (%RSD)		Spike amount (µg/ml)	Accuracy
		Intraday	Inter-day		Recovery (%)
Madecassoside	1.5625	1.47	2.22	1.5625	93.68 ± 0.72
	12.5	0.33	1.41	12.5	93.03 ± 0.44
	25	0.45	0.98	25	98.19 ± 0.04
Asiaticoside	1.5625	0.84	1.87	1.5625	109.40 ± 1.64
	12.5	0.15	1.36	12.5	97.88 ± 2.55
	25	0.16	0.91	25	98.74 ± 3.11
Madecassic acid	1.5625	0.66	2.20	1.5625	105.08 ± 0.55
	12.5	1.05	0.62	12.5	102.39 ± 0.08
	25	1.07	1.18	25	104.91 ± 0.04
Asiatic acid	1.5625	0.15	1.76	1.5625	93.30 ± 0.36
	12.5	0.18	0.64	12.5	104.66 ± 0.10
	25	0.06	0.52	25	107.01 ± 0.06

Condition	Yield (%w/w)	Content (%w/w)
		Madecassoside	Asiaticoside	Madecassic acid	Asiatic acid
1	3.30	0.475 ± 0.002	0.096 ± 0.001	0.039 ± 0.000	0.022 ± 0.000
2	4.55	0.818 ± 0.004	0.166 ± 0.001	0.028 ± 0.000	0.005 ± 0.001
3	3.25	0.810 ± 0.017	0.156 ± 0.002	0.025 ± 0.000	0.008 ± 0.000
4	5.20	0.904 ± 0.006	0.182 ± 0.001	0.037 ± 0.000	0.014 ± 0.000
5	3.20	0.661 ± 0.060	0.151 ± 0.016	0.020 ± 0.001	0.005 ± 0.001
6	4.65	0.941 ± 0.005	0.192 ± 0.000	0.034 ± 0.000	0.008 ± 0.001
7	2.20	0.617 ± 0.052	0.127 ± 0.013	0.018 ± 0.001	0.007 ± 0.000
8	3.75	0.856 ± 0.080	0.169 ± 0.018	0.034 ± 0.002	0.011 ± 0.001
9	2.30	0.623 ± 0.059	0.122 ± 0.013	0.020 ± 0.001	0.006 ± 0.001
10	2.90	0.743 ± 0.002	0.151 ± 0.000	0.032 ± 0.000	0.012 ± 0.000

Polynomial term	Madecassoside		Asiaticoside		Madecassic acid		Asiatic acid
	Coefficient	p-value	Coefficient	p-value	Coefficient	p-value	Coefficient	p-value
Model	-	<0.0001^a a Significant value.	-	<0.0001^a a Significant value.	-	0.0020^a a Significant value.	-	0.0003^a a Significant value.
Intercept	0.6833	-	0.1364	-	0.0286	-	-.0097	-
X₁: temp.	0.1041	<0.0001^a a Significant value.	0.0192	<0.0001^a a Significant value.	0.0026	0.0384^a a Significant value.	-0.0008	0.3373
X₂: time	0.0947	<0.0001^a a Significant value.	0.0168	<0.0001^a a Significant value.	0.0022	0.0770	0.0001	0.8674
X ₁ X ₂	-0.0624	0.0002^a a Significant value.	-0.0109	0.0096^a a Significant value.	0.0058	0.0024^a a Significant value.	0.0056	<0.0001^a a Significant value.
X ₁ ²	0.0547	0.0005^a a Significant value.	0.0153	0.0003^a a Significant value.	-	-	-	-
X ₂ ²	0.0224	0.1086	0.0034	0.3596	-	-	-	-
R ²	0.9042	-	0.8252	-	0.4278	-	0.5101	-
Adjusted R²	0.8843	-	0.7888	-	0.3618	-	0.4536	-
Predicted R²	0.8543	-	0.7315	-	0.2376	-	0.3401	-
Adequate precision	19.7686	-	14.3129	-	7.7524	-	9.1687
Lack of Fit	-	0.6034	-	0.0675	-	<0.0001^a a Significant value.	-	<0.0001^a a Significant value.

Compound	Model	Coded equation
Madecassoside	Quadratic	$Y_{1} = 0.6833 + 0.1041 (X_{1}) + 0.0947 (X_{2}) - 0.0624 (X_{1}) (X_{2}) + 0.0547 {(X_{1})}^{2} + 0.0224 {(X_{2})}^{2}$
Asiaticoside	Quadratic	$Y_{2} = 0.1364 + 0.0192 (X_{1}) + 0.0168 (X_{2}) - 0.0109 (X_{1}) (X_{2}) + 0.0153 {(X_{1})}^{2} + 0.0034 {(X_{2})}^{2}$
Madecassic acid	2FI	$Y_{3} = 0.0286 + 0.0026 (X_{1}) + 0.0022 (X_{2}) + 0.0058 (X_{1}) (X_{2})$
Asiatic acid	2FI	$Y_{4} = 0.0097 - 0.0008 (X_{1}) + 0.0001 (X_{2}) + 0.0056 (X_{1}) (X_{2})$

Response	Predicted value (%w/w)	Actual value (%w/w)	Error (%)^a a %Error = (actual value - predicted value) × 100/actual value.	95% CI (lower-upper)
Y ₁	0.993	0.855 ± 0.0005	-16.1	0.854-1.132
Y ₂	0.203	0.174 ± 0.0001	-16.7	0.165-0.240
Y ₃	0.047	0.053 ± 0.0001	11.3	0.032-0.062
Y ₄	0.020	0.025 ± 0.0003	20.0	0.010-0.030

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[1] * Corresponding author. E-mail:chaowalit@rsu.ac.th (C. Monton).