Determination of nucleosides in Cordyceps sinensis and Ganoderma lucidum by high performance liquid chromatography method

摘要

Background: Nucleosides are supportive in the regulation and modulation of various physiological processes in body, they acts as precursors in nucleic acid synthesis, enhance immune response, help in absorption of iron and influence the metabolism of fatty acids. Cordyceps sinensis and Ganoderma lucidum are well-known for its use in traditional medicine of China, Nepal and India. They are rich in nucleosides such as adenine, adenosine, cordycepin, etc. Hence, a simple, economic and accurate high-performance liquid chromatography (HPLC) analytical method was proposed for determination of adenine and adenosine for the quality control of plants. Materials and Methods: Chromatographic experiments were conducted on YL9100 HPLC system (South Korea). Reversed-phase chromatography was performed on a C18 column with methanol and dihydrogen phosphate as the mobile phase in isocratic elution method at a flow rate of 1.0 mL/min. Detection was carried out at 254 nm, which gives a sharp peak of adenine and adenosine at a retention time of 6.53 ± 0.02 min and 12.41 ± 0.02, respectively. Results and Discussion: Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 25–200 μg/mL for adenosine and 100–800 μg/mL for adenine with regression coefficient of 0.999 and 0.996, respectively. The adenine was found 0.16% and 0.71% w/w in G. lucidum and in C. sinensis , respectively, and adenosine was found to be 0.14% w/w in G. lucidum whereas absent in C. sinensis . Conclusion: The developed HPLC method for the quantification of adenosine and adenine can be used for the quality control and standardization of crude drug and for the different herbal formulations, in which adenine and adenosine are present as major constituents. The wide linearity range, sensitivity, accuracy, and simple mobile phase imply the method is suitable for routine quantification of adenosine and adenine with high precision and accuracy. KEY WORDS: Adenine, adenosine, Cordyceps sinensis , Ganoderma lucidum , high performance liquid chromatography Nucleosides are supportive in the regulation and modulation of various physiological processes in body, they act as precursors in nucleic acid synthesis, enhance immune response, help in absorption of iron and influence the metabolism of fatty acids.[ 1 ] In India, the folk healers of Sikkim use Cordyceps sinensis for the prevention of ailments such as cancer, asthma, tuberculosis, diabetics, cough, cold, and hepatitis.[ 2 ] This C. sinensis is reportedly effective in renal and cardiovascular diseases,[ 2 , 3 , 4 , 5 ] immunological disorders and stress,[ 6 ] and inhibits inflammation and airway hyper-responsiveness. Ganoderma lucidum , is one of the most well-known traditional Chinese herbal medicinal, mushroom-like fungi, has been reported as a potential preventive agent for human disorders by exhibiting its anti-allergy, anti-inflammatory and antioxidant properties.[ 7 , 8 , 9 ] Both the drugs are rich in nucleosides such as adenine, adenosine, cordycepin, etc. As an endogenous nucleoside, adenosine has been widely investigated in different products, for instance, an adenosine is an important index for quality assessment of G. lucidum and C. sinensis .[ 10 , 11 ] Hence, a simple, economic and accurate high-performance liquid chromatography (HPLC) method was proposed for determination of adenine and adenosine for the quality control of both the plants.