Quantitative HPLC Analysis of Benzene Derivatives of Melicope Ptelefolia Leaves

Figures & data

Figure 1 Structures of the three marker benzene derivatives. (Description: The chemical structures of the three markers identified as tHPA, tHGA, and GCA.)

Figure 2 HPLC chromatogram of marker compounds in 1 mg/mL ethanolic leaf extract of M. ptelefolia (tHPA [R_T: 6.25 min], tHGA [R_T: 17.35 min], and GCA [R_T: 20.84 min]). R_T = retention time. (Description: Under the optimized HPLC conditions for the ethanolic M. ptelefolia leaf extract, the best resolution of the three marker components: tHPA, tHGA, and GCA in 1 mg/mL of crude extract was obtained.)

Figure 3 Overlay of UV spectra of the maker compounds in the sample and standards at the same retention time: (a) tHPA, (b) tHGA, and (c) GCA. (Description: Specificity of the method was evaluated by comparing ultraviolet (UV) absorption spectra at the same retention times of the eluted sample peaks with those of the pure standards.)

Table 1  Intra- and inter-day precision of HPLC analysis of tHPA, tHGA, and GCA

Standards	Nominal concentration (μg/mL)	Intraday precision^a detected concentration (μg/mL)	RSD (%)	Interday precision^b detected concentration (μg/mL)	RSD (%)
tHPA	10	8.99	2.2	9.21	3.8
	100	99.5	0.7	102.2	3.0
tHGA	10	9.17	2.1	9.10	2.0
	100	100.3	0.7	103.9	2.7
GCA	10	8.91	1.4	9.62	6.6
	100	100.7	0.7	103.6	3.4
^aAnalyzed on the same day (n = 6).
^bAnalyzed on different days (n = 6).

Table 2  Concentrations of tHPA, tHGA, and GCA in 1 mg/mL of ethanolic crude M. ptelefolia sample solution (mean ± SD; n = 3)

Sample No.	tHPA (mg/mL)	RSD (%)	tHGA (mg/mL)	RSD (%)	GCA (mg/mL)	RSD (%)
1	0.031 ± 0.0003	1.005	0.053 ± 0.0005	0.962	0.209 ± 0.0020	0.965
2	0.029 ± 0.0012	4.079	0.103 ± 0.0034	3.351	0.140 ± 0.0042	2.982
3	0.025 ± 0.0008	3.235	0.087 ± 0.0023	2.700	0.189 ± 0.0049	2.611
4	0.022 ± 0.0007	3.197	0.070 ± 0.0015	2.149	0.150 ± 0.0024	1.606

Table 3  Content of the three marker compounds (w/w) in the leaf crude extract

Sample No.	Crude extract of leaf (g)	Crude^a extract (% yield)	tHPA (g)	Content (%)	tHGA (g)	Content (%)	GCA (g)	Content (%)
1	37.22	9.3	1.15	2.7	1.97	5.3	7.78	20.9
2	69.16	17.3	2.01	2.9	7.12	10.3	9.68	14.0
3	48.62	12.2	1.22	2.5	4.23	8.7	9.19	18.9
4	56.31	14.1	1.24	2.2	3.94	7.0	8.45	15.0
^aThe % yield obtained from each sample was obtained from 400 g of fresh leaves.