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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (11): 1057-1060.doi: 10.3736/jcim20091107

• Original Experimental Research • Previous Articles     Next Articles

Determination of isoguanosine in Semen Crotonis Tiglii by high-performance liquid chromatography

Yun Ling , Hong-yu Zhao, Xin-liang Pan, Chun-xiang Zhou   

  1. Division of Shanghan and Jingui, Preclinical Medicine College,Nanjing University of Chinese Medicine, Nanjing 210046, Jiangsu Province, China
  • Received:2009-06-04 Accepted:2009-09-07 Online:2009-11-20 Published:2009-11-15
  • Contact: Chun-xiang Zhou E-mail:chunxiangzhou@hotmail.com

Objective

To develop a high-performance liquid chromatographic method for the determination of isoguanosine in Semen Crotonis Tiglii.
Methods

The determination was done by using reverse-phase high-performance liquid chromatography with a Hedera ODS-2 column (4.6 mm ×250 mm, 5 μm). Elution was employed with the mobile phase of acetonitrile-methanol-water (1︰4︰95) at flow rate of 1.0 mL/min. The detection wavelength was set at 292 nm and the column temperature was 25 ℃. Injection volume was 10 μL.
Results

The linear range of isoguanosine was from 0.161 to 0.967 mg. The correlation coefficient of the calibration curves was 0.999 8. The average recovery rate was 98.78% with relative standard deviation of 0.02%. Conclusion: The results show that the method is simple, accurate, and repeatable, and it is suitable for quality control of Semen Crotonis Tiglii.

Key words: High-performance liquid chromatography, Semen Crotonis Tiglii, Isoguanosine

Table 1

Recovery rates of isoguanosine"

Semen Crotonis
Tiglii (g)
Assayed
isoguanosine (mg)
Added reference
substance amount (mg)
Detected
amount (mg)
Recovery
rate (%)
Average
recovery rate
RSD
0.3 1.613 2 1.612 3.198 6 98.35
0.3 1.619 7 1.612 3.176 3 96.56
0.3 1.615 4 1.612 3.258 8 101.95 98.78% 0.02%
0.3 1.617 5 1.612 3.228 4 99.93
0.3 1.615 4 1.612 3.163 7 96.05
0.3 1.613 2 1.612 3.222 7 99.84

Table 2

Determination of isoguanosine in Semen Crotonis Tiglii ($\bar{x}$±s,mg/g)"

No. Habitat n Batch No. Concentration
1 Yunnan 3 20080906 12.0±1.4
2 Yunnan 3 20081145 11.9±1.8
3 Yunnan 3 20081028 16.2±0.5
4 Sichuan 3 080301 13.4±1.4
5 Guangxi 3 080815 10.8±1.8
6 Sichuan 3 080816 11.2±1.5
7 Sichuan 3 080817 11.7±0.4
8 Sichuan 3 081206 12.4±1.8
9 Yunnan 3 090205 10.2±0.6
10 Gansu 3 090210 10.6±1.7

Figure 1

HPLC chromatograms of reference substance and sample A: Reference substance; B: Sample; 1: Isoguanosine."

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