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Sedative Effect and Standardization Parameters of Herbal Medicinal Product Obtained from the Ocimum americanum L. Herb

M. Shanaida
M. Shanaida
, 
O. Golembiovska
O. Golembiovska
, 
I. Jasicka-Misiak
I. Jasicka-Misiak
, 
O. Oleshchuk
O. Oleshchuk
, 
N. Beley
N. Beley
, 
I. Kernychna
I. Kernychna
 et 
P.P. Wieczorek
P.P. Wieczorek
   | 14 avr. 2021
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Article Category: Original Paper
Publié en ligne: 14 avr. 2021
Pages: 1 - 9
Reçu: 09 sept. 2020
Accepté: 01 févr. 2021
DOI: https://doi.org/10.2478/afpuc-2020-0015
Mots clés
© 2021 Shanaida M. et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

HPTLC chromatograms of the OATEs (1), Ocimum americanum essential oil (2), and linalool standard (A) after the derivatization with anisaldehyde solution at day light. Mobile phase: ethyl acetate – toluene (5:95).
HPTLC chromatograms of the OATEs (1), Ocimum americanum essential oil (2), and linalool standard (A) after the derivatization with anisaldehyde solution at day light. Mobile phase: ethyl acetate – toluene (5:95).

Figure 2

HPTLC chromatograms of the OATEs (A) and polyphenols standards (1 – rutin; 2 – caffeic acid; 3 – RA) after the derivatization with 1% methanolic AlCl3 at λ=366 nm. Mobile phase: ethyl acetate – formic acid – water (15:1:1).
HPTLC chromatograms of the OATEs (A) and polyphenols standards (1 – rutin; 2 – caffeic acid; 3 – RA) after the derivatization with 1% methanolic AlCl3 at λ=366 nm. Mobile phase: ethyl acetate – formic acid – water (15:1:1).

Figure 3

UV absorption spectrum of RA in the HPLC method
UV absorption spectrum of RA in the HPLC method

Figure 4

HPLC chromatogram of the RA standard solution (A) and OATEs (B) (λ=330 nm)
HPLC chromatogram of the RA standard solution (A) and OATEs (B) (λ=330 nm)

The robustness testing of RA identification in OATEs by HPTLC.

Indicator Level of changes Rf variability (RSD, %)
Mobile phase composition (ethyl acetate – formic acid – water, V/V) 15:1:1 0.8
15:1.05:1 1.3
15:0.95:1 1.7
15:1:1.05 1.3
15:1:0.95 1.3
Development distance, cm 8 0.8
18 1.3
Chamber 3 different containers 1.7
Analysts 2 different analysts 0.6

The system suitability parameters for the determination of RA by HPLC.

Standard Retention time (Rt) Tailing factor (T) Theoretical plates (N)
Rosmarinic acid 38.0 1.2 2399586

The gradient mode in the HPLC analysis.

Time (min) Mobile phase A (vol%) Mobile phase B (vol%)
0–5 95 5
5–35 95 → 75 5 → 25
35–40 75 25
40–60 75 → 50 25 → 50
60–65 50 → 20 50 → 80
65–70 20 80
70–85 95 5

The linearity, regression equation, limit of detection, limit of quantitation, precision parameters of RA standard in the HPLC analysis.

Standard Linearity range [μg/ml] Regression equation R2 LOD [μg/ml] LOQ [μg/ml] RSD [%]
Inter-day Intra-day
Rosmarinic acid 5–50 Y = 0.235x + 0.164 0.9998 0.005 0.015 0.16 0.04

The results of HPTLC analyses of biologically active compounds in OATEs.

Compound Rf value Colour Presence in OATEs Presence in essential oil of O. americanum
Method A
Unknown compound 0.03 Violet + −
Unknown compound 0.11 Violet + −
Unknown compound 0.19 Violet + −
Linalool 0.29 Violet + +
Unknown compound 0.85 Violet + +
Method B
Rutin 0.09 Yellow + *
Unknown compound 0.14 Blue + *
Unknown compound 0.27 Yellow + *
Unknown compound 0.29 Blue + *
Unknown compound 0.43 Dark blue + *
Unknown compound 0.64 Blue + *
Rosmarinic acid 0.75 Blue + *
Caffeic acid 0.79 Blue + *

The influence of tinctures prepared from Ocimum americanum herb on the spontaneous behavior of rats in the open field test (n = 6).

Group Number of squares crossed Tentative research activity Indicators of anxiety (emotional reactions)
Vertical racks Examined holes Defecations Urinations Grooming
Control 32.61 ± 2.29 9.52 ± 0.89 5.21 ± 0.47 1.03 ± 0.1 0.51 ± 0.05 2.32 ± 0.21
OAT 29.17 ± 1.09 8.83 ± 0.28 4.83 ± 0.35 0.67 ± 0.091 0.5 ± 0.05 2.33 ± 0.24
OATEs 26.0 ± 1.021 8.0 ± 0.421 4.5 ± 0.21 0.5 ± 0.051 0.33 ± 0.021,2 2.17 ± 0.19
Diazepam 18.5 ± 0.541 6.5 ± 0.181 3.17 ± 0.091 0.5 ± 0.021 01 1.17 ± 0.041

The robustness testing for the identification of linalool in OATEs by HPTLC.

Indicator Level of changes Rf variability (RSD, %)
Mobile phase composition (ethyl acetate – toluene, V/V) 5:95 0.6
5.2:95 1.3
4.8:95 0.8
Development distance, cm 8 1.3
18 1.8
Chamber 3 different containers 1.3
Analysts 2 different analysts 0.8
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