| Ganoderma lucidum is the dried fruiting body of Ganoderma lucidum (Leyss.ex Fr.)Karst. or Ganoderma sinense Zhao,Xu et Zhang, which has long been used in traditional chinese medicine f-or the promotion of longevity and maintenance of vitaliy. Zhiji Capsule is one kind of health foods which is made of G. lucidum and has the function of adjusting immunity and so on.The present stu-dy provides the methods to isolate and simultaneously determine the major biological components in G. lucidum and Zhiji Capsule and HPLC fingerprint chromatogram was also established for the herb. Subsequently, the systematic quality evaluation of G.lucium and its capsule were developed.Triterpenic acids have been reporting as effective components of G.lucidum. The fruiting bodies of G.lucidum were extracted with 95% aqueous alcohol to give an alcoholic extract, which was extracted with dichlormethane,dissolved by base and precipitated by acid to afford the total triterpenic acids. Seven compounds were isolated from the total triterpenic acids by silica gel, sephadex LH-20 and pre-HPLC methods and they were identified as follow by chemical reactions and spectral analysis:ganoderic acid DM,lucidone A,ganosporelactone B,methyl ganoderate A,ganoderic acid D, ganoderenic acid A and ganoderic acid A.The RP-HPLC fingerprint analysis for the quality control of G.lucidum was established. The separation was performed on a Kromasil C18 analytical column (250 mm×4.6 mm,5μm) with the mobile phase consisting of acetonitrile(A) and 0.03% aqueous phosphonic acid(B) with gradient elution mode at the flow rate of 1.0 mL-min-1. Elution procedure:0-35 min,32%-33%A; 35-38 min,33%-37%A; 38-60 min,37%-40%A; 60-70 min,40% A. The wavelength of measurement was 252 nm and the column temperature was set at 35℃.16 batches of G.lucidum were determined under the condition described above. With the Within-groups linkage method and the Cosine used as the measurement index, the Hierarchical cluster was applied to the fingerprints of G.lucidum and the 16 batches of G.lucidum was divided into two grades. Among them the grade I was commendatory and the grade II was non-commendatory. Mutual pattern was established from 11 batches in the grade I and the similarity data were also obtained. The results of the commendatory should be all over 0.90 and the other samples were the non-commendatory.A reversed-phase high performance liquid chromatographic method was developed for the simultaneous determination of four triterpenic acids namely ganoderic acid C2, ganoderenic acid A, ganoderic acid A, ganoderic acid D in 16 batches of G. lucidum. The separation was performed on Kromasil C18 column (250 mm×4.6 mm,5μm) with the mobile phase consisting of acetonitrile(A) and 0.03% aqueous phosphonic acid(B) with gradient elution mode at the flow rate of 1.0 mL·min-1. Elution procedure:0-35 min,32%-33%A; 35-38 min,33%-37%A; 38-60 min,37%-40%A; 60-65 min,40% A. The wavelength of measurement was 252 nm and the column temperature was set at 35℃. The calibration curves of ganoderic acid C2, ganoderenic acid A, ganoderic acid A, ganoderic acid D were linear over the range of 5.0-50.0μg·mL-1(r= 0.9999),7.2-72μg·mL-1(r= 0.9999),11.67-116.7μg·mL-1(r=0.9999) and 5.32-53.2μg·mL-1(r= 0.9998), respectively. The mean recoveries were 98.7%(RSD=1.8%),99.1%(RSD=2.1%),99.6%(RSD=1.9%) and 98.5%(RSD=1.8%), respectively.7 batches of Zhiji Capsules were analyzed under the same chromatographic condition. The mean recoveries were 98.6%(RSD=1.8%),100.7%(RSD=2.2%), 99.5%(RSD=2.1%) and 99.0%(RSD=1.8%).The assay was found to be simple simple, accurate and reproducible, it can be used for the quality control of G. Lucidum and Zhiji Capsule. |
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