The Study Of Petroleum Ether Extract Of Cistanche Deserticola
Mar 14, 2022
Chemical constituents and cell cycle inhibitory activity of petroleum ether extract of Cistanche deserticola
Contact: joanna.jia@wecistanche.com / WhatsApp: 008618081934791
Lu Kang, Liu Hongbing, Gu Qianqun*
(Key Laboratory of Marine Medicine, Ministry of Education, Institute of Marine Medicine and Food, Ocean University of China, Qingdao, Shandong 266003)
Cistanche deserticola Y. C. Ma, also known as Cistanche, is a plant belonging to the genus Cistanche in the family Lydanaceae. Its dry fleshy stem with scales is used as a traditional Chinese medicine Cistanche deserticola. It is a genuine product included in the Pharmacopoeia of the People's Republic of China. Cistanche is sweet in taste and warm in nature. It has the effect of "tonifying kidney yin, nourishing essence, and blood, moisturizing the intestines and laxative". It can be used to treat male impotence, female infertility, cold waist and knee pain, blood dry constipation, etc. [1].
In recent years, the traditional Chinese medicine Cistanche has attracted people's attention because of its remarkable memory improvement, anti-aging, anti-fatigue, and enhancement of body immunity. Its chemical composition and pharmacological effects have been reported more [2, 3]. However, literature search shows that there are few related studies on fresh desert Cistanche, only Tu Pengfei and others have studied the chemical composition of the n-butanol extract of its flowers [4]. In the systematic study of the chemical components of fresh Cistanche, the author found that both petroleum ether extract and ethyl acetate extract have certain anti-tumor activities. In this experiment, GC-MS was used to analyze and identify the chemical composition of petroleum ether extract, and flow cytometry was used to detect its cycle inhibitory activity on mouse breast cancer ts FT210 cells.
1 Experimental part
1. 1 Instruments, reagents, and samples
GC-M S combined instrument British V G company, Fiso ns M D800GC /M S type. Using mouse breast cancer ts-FT210 cells. Flow cytometry uses Va n-tage type flow cytometer from BD company in the United States, and the distribution of cells in each phase of the cell cycle is analyzed and calculated by Mod Fit LT software. Fetal Bovine Serum (FBS) and RPM I-1640 cell culture medium were produced by Hyclone (Cat. No. STF721) and Gibcobrl, respectively. Propidium iodide (PI) and Triton X-100 were both produced by Sigma. The new fresh cistanche medicinal material was collected in Alxa League, Inner Mongolia, and was identified as a fleshy stem with scale leaves of cistanche C. deserticola Y. C. Ma by Mr. Liu Hongbing from the Institute of Marine Medicine and Food Research of the Ocean University of China.
1. 2 Extraction of samples
After 450 g of fresh desert Cistanche, medicinal material was chopped, it was diafiltered with 2 000 mL petroleum ether with a boiling range of 30～60℃, the solvent was recovered under low temperature and reduced pressure, and then dried over anhydrous sodium sulfate to obtain a yellow transparent liquid, which was sealed and stored at -10 Keeps in the refrigerator at ℃. The yield was 0.93%.
1.3 GC-MS analysis of petroleum ether extract
Gas chromatographic conditions: Chromatographic column is 30QC2/AC5 quartz capillary column (30 m×0.32 mm); column temperature is 80～280℃, the programmed temperature rise is 3℃/min; split ratio is 30:1; vaporization chamber temperature is 240℃; The front pressure of the column is 82.8 kPa; the carrier gas is high-purity N2.
Mass spectrometry conditions: EI-MS, electron energy 70 eV; ion source temperature 200°C; filament current 4.1 A; mass range 35～450 m/z; scanning period 1 s; LAB-BASE system for data processing, using the United States The NBS library of the National Bureau of Standards was searched and confirmed by reference.
1. 4 Anti-tumor activity test
Refer to literature method [5], using flow cytometry to detect the cycle inhibitory activity of Cistanche deserticola petroleum ether extract on tsFT210 cells.
2. Results
2. 1 Identification of the chemical composition of petroleum ether extract
After analysis and identification by GC-MS combined instrument, a total of 46 peaks were detected in the petroleum ether extract of Cistanche deserticola, which was searched with the NBS library of the National Bureau of Standards and confirmed by reference, and a total of 25 compounds were identified, accounting for the peak area. Of 75. 03%, the results are shown in Table 1. The results showed that esters and alkanes are the main components of Cistanche petroleum ether extract. The highest mass fraction is 14-methyl-pentadecanoic acid methyl ester (13.61%), followed by hexadecanoic acid ethyl ester (12.39%) 2,5,6-trimethyl-octane. Alkane (7.60%).
2.2 Anti-tumor activity
The results of flow cytometry showed that the petroleum ether extract of Cistanche deserticola had obvious G0/G1 phase inhibitory activity on tsFT210 cells at 10μg/mL (Figure 1).
Table 1 Constituents of petroleum ether extract from fresh C. deserticola
3. Discussion
At present, the research on the biological activity of Cistanche is mainly focused on improving memory, anti-aging, and enhancing the immune function of the body. There is no report on the anti-tumor aspect. In this experiment, flow cytometry was used to preliminarily evaluate the cell cycle inhibitory activity of fresh Cistanche deserticola petroleum ether extract on mouse breast cancer tsFT210 cell cycle, and use GC-MS to analyze its chemical composition. Among the petroleum ether extracts of Cistanche deserticola, there are mainly alkanes and alkanoate esters and a small number of phenols and cyclohexanone compounds. The anti-tumor activity of these compounds has not been reported yet. Therefore, the anti-tumor active ingredients in the petroleum ether extract of Cistanche deserticola need further research to determine.
Acknowledgments: GC-MS was determined by Professor Ding Jingkai, Kunming Institute of Botany.
References
[1] Jiangsu New Medi cal Coll ege. Dictionary y of Chinese Ma teria Med ica (Chinese Medicine Dictionary) [M ]. Sh shanghai: Shanghai People e's Publishing House, 1997.
[2] Song ZH, Lei L, Tu P F. Advances in research of pharma- col logical activity in plants of Cistanche Huffing. et Link [J]. Chin Tra dit Herb Drugs (Chinese herbal medicine), 2003, 34( 9): 16-17.
[3] Zhang LH, Du N S. Advances in research of chemical constituents in plants of Herba Cistanche [J]. Chin Trad it Pat Med (Chinese patent medicine), 2003, 25( 4): 323-327.
[4] Tu PF, He YP, Lou Z C. Studies on the chemical constituents of the fresh inflorescences of d desert living cist Anche (Cistanche deserticola) [J]. Chin Trad it Herb Drugs (Chinese herbal medicine ), 1994, 25(9): 451-452.
[5] Liu HB, Cui CB, Cai B, et al. Isolation, identification and antitumor activity of triterpenes from Pterocar ya tonkinesis (French.) Dode [J]. Chin J Med Chem (Chinese Journal of Medicinal Chemistry ), 2004, 14(3): 168-170.
From: Chinese Traditional and Herbal Drugs; Volume 36 Number 4 April 2005.
