Abstract:
An analytical method for identifying woods-grown ginseng and mountain cultivated ginseng comprise: (a) preparing ginseng extracts, (b) acquiring fingerprint regions by GC-MS analysis, (c) selecting at least 8 GC peaks for similarity calculation; (d) calculating similarity coefficients; and (e) comparing similarity coefficients of samples to those of standards.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a continuation of International Patent Application No. PCT/CN2006/002319 with an international filing date of Sep. 7, 2006, designating the United States, now pending, and further claims priority benefits to Chinese Patent Application No. 200510030084.4, filed on Sep. 28, 2005. The contents of these specifications are incorporated herein by reference. 
    
    
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     This invention relates to analytical methods, and particularly to analytical identification methods applicable to identification of woods-grown ginseng and mountain-cultivated ginseng. 
     2. Description of the Related Art 
     Radix ginseng is the dried root of a perennial plant Panax ginseng C. A. Meyer (Araliaceae). Panax ginseng C. A. Meyer (oriental ginseng) comes in at least three varieties depending on where and how it is grown: wild ginseng, woods-grown ginseng, and mountain-cultivated ginseng (also known as garden ginseng). All of these varieties derive from the same root source. 
     Wild ginseng grows without human interference in remote mountains and dense forests under the conditions of natural climate, soil, slope, forest canopy, moisture and associated plants. It is a perennial, rare and endangered plant, listed as the first class of national protection species in the  Chinese Red Book,  1 st  Ed. Wild Panax ginseng is not allowed to be collected and traded in China. 
     Woods-grown ginseng, seeded artificially, grows in undisturbed natural forest for 15-20 years or longer. The cultivation takes advantage of fertile soil, proper temperature, light and water in a broadleaf forest or a mixed forest of broadleafs and conifers, which imitates the natural growth conditions of wild Panax ginseng. Woods-grown ginseng can be induced to have some of the same characteristics of wild Panax ginseng. Because wild Panax ginseng is increasingly scarce, woods-grown ginseng is the next best thing and is universally sought-after for its medicinal properties. 
     Mountain-cultivated ginseng is planted in a suitable, hilly area after the area has been logged. Mountain-cultivated ginseng is typically picked after 5-6 years of growth. 
     The length of cultivation and growth conditions determine ginseng characteristics. Wild ginseng is better in terms of its medicinal properties than wood-grown ginseng, which is in turn better than mountain-cultivated ginseng. However, differentiation of ginseng varieties is difficult based on their exterior characteristics or the characteristics of ground ginseng powder. 
     The exterior characteristics of ginseng root include the rhizome, the main root, the lateral root, rootlets, and annular marks, which may be used to identify a variety of ginseng. Often, however, exterior characteristics don&#39;t reflect the internal quality of ginseng accurately and overemphasis of the usefulness of exterior characteristics in the identification process leads to loss of the consumer confidence. 
     Chemical characterization work on ginseng has begun as early as in 1939 with a large number of reports appearing in the 1960s. The chemical composition of ginseng extracts is very complicated, with about 200 different compounds having been identified. They are divided into several general classes according to their chemical structures:
         1) Terpenes: including monoterpenes, such as α-pinene, β-pinene, dihydrocarvone, carvone, etc; sesquiterpenes, such as straight chain sesquiterpenes nerolidol, cis-farnesene, trans-β-farnesene, etc, single ring sesquiterpenes, such as β-bisabolene, α-elemene, β-elemene, etc; bicyclic sesquiterpenes, such as α-selinene, β-selinene, β-caryophyllene, β-santalol, etc.   2) Aliphatic compounds, including alkanes, alkenes, alkynes, alcohols, ethers, aldehydes, ketones, acids, esters etc.   3) Aromatic compounds, such as ethylbenzene, tert-butylbenzene, 1,2-dimethyl benzene, naphthalene, naphthylamine, etc.; and   4) Heterocyclic compounds, such as 2-pentylfuran, 5-methyl-2-furfural, benzofuran, teramethyl pyrazine, 1,2-dimethyl anthraquinone, etc.       

     The amount and contents of various compounds in fresh ginseng extracts vary depending on the length of ginseng growth period and cultivation area, which makes it possible to identify different varieties of ginseng by chromatographic analysis. 
     The concept of chromatographic fingerprint analysis of complex mixtures is known. By comparing chromatographic fingerprints, samples can be identified. Chromatographic fingerprint analysis involves comparing the similarities and differences (relative intensities) of the corresponding peaks of each chromatogram using a quantization process. Chromatographic fingerprint analysis comprises the following steps: (1). Digitization of the fingerprint (peak measurement, e.g. peak area, peak height etc.); and (2) Calculation of fingerprint spectrum similarity. 
     In the early research on chromatogram fingerprints, the digitization was realized using the peak height or peak area extracted from the main peak of a chromatogram. However, this method used part of the chromatogram characteristics to express the whole fingerprint, discarding certain information and at last reducing the identification sensitivity of chromatogram fingerprint. 
     Spectra detected by the modern chromatography are stored as a group of binary numbers in the computer. This group of numbers is called a vector, which conveys the holographic information of the entire spectrum. The similarity measured by this kind of vector reflects the similarity of the entire chromatographic fingerprint of the chemical species. Any chemical species can be expressed by a vector. The resemblance between two chemical species can be measured by the similarity between their vectors. 
     Chromatogram fingerprint similarity can be calculated from one, several, or all peaks in the chromatogram fingerprint. The latter fingerprint similarity is usually used to measure the overall fluctuation of chemical composition in crude drugs. 
     Spectra obtained by modern chromatography are usually stored as a group of binary numbers, i.e., a vector. A similarity coefficient S between two chromatographic spectra can be calculated by the following formula: 
     
       
         
           
             S 
             = 
             
               
                 
                   ∑ 
                   
                     i 
                     = 
                     1 
                   
                   n 
                 
                 ⁢ 
                 
                   
                     ( 
                     
                       
                         x 
                         i 
                       
                       - 
                       
                         x 
                         _ 
                       
                     
                     ) 
                   
                   ⁢ 
                   
                     ( 
                     
                       
                         y 
                         i 
                       
                       - 
                       
                         y 
                         _ 
                       
                     
                     ) 
                   
                 
               
               
                 
                   
                     
                       ∑ 
                       
                         i 
                         = 
                         1 
                       
                       n 
                     
                     ⁢ 
                     
                       
                         ( 
                         
                           
                             x 
                             i 
                           
                           - 
                           
                             x 
                             _ 
                           
                         
                         ) 
                       
                       2 
                     
                   
                 
                 ⁢ 
                 
                   
                     
                       ∑ 
                       
                         i 
                         = 
                         1 
                       
                       n 
                     
                     ⁢ 
                     
                       
                         ( 
                         
                           
                             y 
                             i 
                           
                           - 
                           
                             y 
                             _ 
                           
                         
                         ) 
                       
                       2 
                     
                   
                 
               
             
           
         
       
     
     In this formula,  x  and  y  respectively represent the mean values of two groups of chromatographic numbers and x i  and y i  represent the corresponding points of chromatographic numbers. The obtained related coefficient is Sε[ 0 , 1 ]. Thereby, a similarity between the two groups of numbers can be determined. The similarity gained by this method can reflect the resemblance of the entire spectra of two samples. 
     Gas chromatography (GC) uses gas as the mobile phase. It separates components according to their partition ratios in the immobile phase and gas-carrying phase, which has the advantages of high resolution, good selectivity, rapid measurement and wide applicability. Accordingly, GC is particularly useful in determining relative composition of plant extracts and other volatile mixtures. 
     GC-MS is a technique which combines mass spectroscopy with gas chromatography. This coupling technique records mass spectroscopic data of various compounds of mixture after they have been resolved by GC. This allows for an easier identification of each peak in a chromatographic fingerprint. 
     SUMMARY OF THE INVENTION 
     In one embodiment, this invention provides a method for identifying between woods-grown ginseng and mountain-cultivated ginseng, comprising: (a) preparing a ginseng extract of a ginseng sample; (b) obtaining a GC-MS spectrum of the extract; (c) calculating a similarity coefficient based on at least 8 peaks in the GC-MS spectrum with respect to a selected standard; and (d) comparing the similarity coefficient calculated in (c) to pre-calculated standard similarity coefficients. 
     In a class of this embodiment, the standard similarity coefficients are: for woods-grown ginseng: main root 0.88-0.91 median mode, 0.77-0.88 mean value mode; lateral root 0.78-0.96 median mode, 0.78-0.96 mean value mode; fibrous root 0.81-0.96 median mode, 0.90-0.92 mean value mode; for mountain cultivated ginseng: main root 0.38-0.57 median mode, 0.36-0.54 mean value mode; lateral root 0.24-0.57 median mode, 0.24-0.58 mean value mode; fibrous root 0.25-0.61 median mode, 0.24-0.71 mean value mode. 
     In another class of this embodiment, the GC-MS spectrum is obtained at a column temperature rising from 100° C. to 170° C. at 1.5° C./min, then from 170° C. to 190° C. at 8.0° C./min and at last from 190° C. to 240° C. at 2.0° C./min. 
     In another class of this embodiment, the peaks are common to woods-grown ginseng and mountain-cultivated ginseng samples. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  illustrates a GC-MS chromatogram of ginseng extracts from main root of woods-grown ginseng; 
         FIG. 2  illustrates a GC-MS chromatogram of ginseng extracts from main root of mountain-cultivated ginseng; 
         FIG. 3  illustrates a GC-MS chromatogram of ginseng extracts from main root of 15-year-old woods-grown ginseng; 
         FIG. 4  illustrates a GC-MS chromatogram of ginseng extracts from lateral root of 15-year-old woods-grown ginseng; 
         FIG. 5  illustrates a GC-MS chromatogram of ginseng extracts from rootlets of 15-year-old woods-grown ginseng; 
         FIG. 6  illustrates a GC-MS chromatogram of ginseng extracts from main root of 18-year-old woods-grown ginseng; 
         FIG. 7  illustrates a GC-MS chromatogram of ginseng extracts from lateral root of 18-year-old woods-grown ginseng; 
         FIG. 8  illustrates a GC-MS chromatogram of ginseng extracts from rootlets of 18-year-old woods-grown ginseng; 
         FIG. 9  illustrates a GC-MS chromatogram of ginseng extracts from main root of 4-year-old Jian City mountain-cultivated ginseng; 
         FIG. 10  illustrates a GC-MS chromatogram of ginseng extracts from lateral root of another specimen of 4-year-old Jian City mountain-cultivated ginseng; 
         FIG. 11  illustrates a GC-MS chromatogram of ginseng extracts from rootlets of 4-year-old Jian City mountain-cultivated ginseng; 
         FIG. 12  illustrates a GC-MS chromatogram of ginseng extracts from main root of 6-year-old Jingyu County mountain-cultivated ginseng; 
         FIG. 13  illustrates a GC-MS chromatogram of ginseng extracts from lateral root of 6-year-old Jingyu County mountain-cultivated ginseng; 
         FIG. 14  illustrates a GC-MS chromatogram of ginseng extracts from rootlets of 6-year-old Jingyu County mountain-cultivated ginseng; 
         FIG. 15  illustrates a GC-MS calibration spectrogram of ginseng extracts from main roots of three batches of woods-grown ginseng; 
         FIG. 16  illustrates a GC-MS chromatogram of ginseng extracts from main roots of three batches of woods-grown ginseng after calibration; 
         FIG. 17  illustrates a GC-MS chromatogram of a standard of ginseng extracts from main roots of three batches of woods-grown ginseng (mean value); 
         FIG. 18  illustrates a GC-MS chromatogram of a standard of ginseng extracts from main roots of three batches of woods-grown ginseng (median value); 
         FIG. 19  illustrates a GC-MS chromatogram of ginseng extracts from main roots of three batches of woods-grown ginseng and five batches of mountain-cultivated ginseng after calibration; 
         FIG. 20  illustrates a GC-MS chromatogram of ginseng extracts from lateral roots of three batches of woods-grown ginseng after calibration; 
         FIG. 21  illustrates a GC-MS chromatogram of a standard of ginseng extracts from lateral roots of three groups of woods-grown ginseng (mean value); 
         FIG. 22  illustrates a GC-MS chromatogram of a standard of ginseng extracts from lateral roots of three batches of woods-grown ginseng (median value); 
         FIG. 23  illustrates a GC-MS chromatogram of ginseng extracts from lateral roots of three batches of woods-grown ginseng and five batches of mountain cultivated ginseng after calibration; 
         FIG. 24  illustrates a GC-MS chromatogram of ginseng extracts from rootlets of three batches of woods-grown ginseng after calibration; 
         FIG. 25  illustrates a GC-MS chromatogram of a standard of ginseng extracts from rootlets of three batches of woods-grown ginseng (mean value); 
         FIG. 26  illustrates a GC-MS chromatogram of a standard of ginseng extracts from rootlets of three batches of woods-grown ginseng (median value); and 
         FIG. 27  illustrates a GC-MS chromatogram of ginseng extracts from rootlets of three batches of woods-grown ginseng and five batches of mountain-cultivated ginseng after calibration. 
     
    
    
     DETAILED DESCRIPTION OF THE EMBODIMENTS 
     GC-MS technology is used to determine the chemical composition of fresh ginseng extracts via its chromatographic fingerprint. The chromatographic fingerprints of ginseng extracts from various species of ginseng are used to differentiate woods-grown ginseng from mountain-cultivated ginseng. The method of identification according to the invention is based on the following techniques. 
     1) Preparation of Ginseng Extracts 
     Various protocols for preparing ginseng extracts were tried to maximize extraction efficiency, wherein the following were varied: extraction solvent (n-hexane, ether, acetone), extraction mode (ultrasound, quench), extraction time (30 min, 1 hr, 2 hrs), extraction temperature (room temperature, 30° C.), and purification (centrifugation, liquid-liquid extraction). The following protocol was determined to be most efficient. Main root, lateral root and rootlets of ginseng were separately broken up into pieces. About 50 mg of each sample was placed into a 1 mL Eppendorf tube, and 0.5 mL of n-hexane were added to each. The samples were sonicated for 1 h and centrifuged. The supernatant was decanted and used directly for GC-MS analysis. 
     2) Chromatographic Conditions: 
     Various protocols for GC-MS analysis were tried to maximize resolution, wherein the following were varied: chromatography column, injection volume (1 μL, 3 μL), carrier gas flow rate (1.0, 1.2, 1.3, 1.5 mL/min), and temperature programs (13 various temperature programs). The following protocol resulted in the best resolution: Dikma DM-1 (30 mm×0.25 mm×0.25 um) capillary column; column temperature 100° C.; injection temperature: 270° C.; temperature program (see Table 1); injection volume: 1 μL; injection mode: split stream, split ratio 2.0; flow rate of carrier gas: 1.30 mL/min. 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 GC Temperature Program 
               
             
          
           
               
                 Rate 
                 Temperature 
                 Hold time 
               
               
                 (° C./min) 
                 (° C.) 
                 (min) 
               
               
                   
               
               
                 — 
                 100.0 
                 0.00 
               
               
                 1.5 
                 170.0 
                 0.00 
               
               
                 8.0 
                 190.0 
                 0.00 
               
               
                 2.0 
                 240.0 
                 0.00 
               
               
                   
               
             
          
         
       
     
     Experimental Conditions 
     1) Sample Preparation 
     Ginseng samples were selected by conventional method. n-Hexane was added and the samples were sonicated for 1 hr. The samples were then centrifuged and the supernatant was decanted for GC-MS analysis. 
     2) GC-MS Analysis 
     GC conditions: Dikma DM-1 (30 mm×0.25 mm×0.25 um) capillary column; column temperature: 100° C.; injection temperature: 270° C.; temperature program (Table 1); injection volume: 1 μL; mode of sample injection: split flow, rate of split flow 2.0; flow rate of carrier gas: 1.30 mL/min. 
     MS conditions: ion source temperature: 200° C.; interface temperature: 250°C.; solvent cut time: 5 min; quality scanning area: m/z 35-380; scanning interval: 0.5 s. 
     GC-MS charts of the samples were obtained under the conditions listed above. 
     3) Identification of the Samples 
     Between 8 and 9 GC peaks were chosen in the chromatographic fingerprint region obtained from step 2) to calculate similarity relative to a standard spectrum (mean or median spectrum of main root, lateral root or rootlets of woods-grown ginsengs). 
     Then, calculated values were compared with the following standard similarity values to identify the sample as woods-grown ginseng or as mountain-cultivated ginseng. 
     Woods-Grown Ginseng:
         main root: 0.88-0.91 median mode, 0.77-0.88 typical value mode;   lateral root: 0.78-0.96 median mode, 0.78-0.96 typical value mode;   fibrous root: 0.81-0.96 median mode, 0.90-0.92 typical value mode;       

     Mountain-Cultivated Ginseng:
         main root 0.38-0.57 median mode, 0.36-0.54 typical value mode;   lateral root 0.24-0.57 median mode, 0.24-0.58 typical value mode;   fibrous root 0.25-0.61 median mode, 0.24-0.71 typical value mode.       

     Setting the GC-MS charts of main root, lateral root and rootlets of woods-grown ginseng as the standard of chromatographic fingerprints, by calculating the similarity values between the ginseng sample to be examined and the mutual modes, the identity of a sample (whether woods-grown or mountain-cultivated) can be ascertained. 
     After extracting 50 μg extracts composition of ginseng using a conventional method, the invention uses examination conditions mentioned above to perform a GC-MS analysis, i.e., to make the GC-MS charts of the extracts as vectors to calculate the similarity value of woods-grown ginseng and cultivated ginseng aided by a computer assessment system of similarity. In this way, woods-grown ginseng can be effectively distinguished from mountain-cultivated ginseng. The method can be also used to determine the part of the ginseng from which extract was made. It can also be used for identification of ginseng powder and other ginseng products. 
     EXAMPLES 
     Example 1 
     Identification Between Woods-Grown Ginseng and Mountain Cultivated Ginseng 
     1) Equipment and Agent 
     Shimadzu GC-MS MS-QP2010 gas chromatograph-mass spectrometer 
     Crest ultrasonic cleaning machine (Model 1875 HTAG) 
     Spectrafuge 16M Hydroextractor 
     N-Hexane: Lab-Scan Analytical Sciences, CBAC-34484-2.5 L 
     2) Sample Source 
     All the ginseng samples used in the research were purchased from different areas in Jilin Province. See Table 2. 
     
       
         
               
             
               
               
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Type and Geological Origin of Ginseng Samples 
               
             
          
           
               
                 Sample 
                   
                   
               
               
                 No. 
                 Type 
                 Geological origin 
               
               
                   
               
             
          
           
               
                 1 
                 Woods-grown ginseng (11-year-old) 
                 Linjiang Region 
               
               
                 2 
                 Woods-grown ginseng (15-year-old) 
                 Linjiang Region 
               
               
                 3 
                 Woods-grown ginseng (18-year-old) 
                 Linjiang Region 
               
               
                 4 
                 Woods-grown ginseng (11-year-old) 
                 Linjiang Region 
               
               
                 5 
                 Woods-grown ginseng (15-year-old) 
                 Linjiang Region 
               
               
                 6 
                 Woods-grown ginseng (18-year-old) 
                 Linjiang Region 
               
               
                 7 
                 Mountain cultivated ginseng (6-year-old) 
                 Jian City 
               
               
                 8 
                 Mountain cultivated ginseng (5-year-old) 
                 Jian City 
               
               
                 9 
                 Mountain cultivated ginseng (4-year-old) 
                 Jian City 
               
               
                 10 
                 Mountain cultivated ginseng (5-year-old) 
                 Jingyu County 
               
               
                 11 
                 Mountain cultivated ginseng (6-year-old) 
                 Jingyu County 
               
               
                   
               
             
          
         
       
     
     3) Preparation of Sample Extract 
     Ginseng samples, including main root, lateral root and rootlet samples, were crushed and grinded individually. 50 mg of powder was picked from each sample and sonicated in about 0.5 mL of n-hexane in a 1 mL Eppendorf tube for 60 min. The final solution was centrifuged to obtain clear supernatant liquid for GC-MS analysis. 
     4) Experimental Condition: 
     4.1) GC condition: Dikma DM-1 (30 mm*0.25 mm*0.25 μm) capillary column; column temperature: 100° C.; injection temperature: 270° C.; temperature program (Table 1); injection volume: 1 μL; injection mode: split stream, split ratio 2.0; flow rate of carrier gas: 1.30 mL/min. 
     4.2) MS condition: Ion source temperature: 200° C.; interface temperature: 250° C.; solvent cut time: 5 min; quality scanning area: m/z 35-380; scanning interval: 0.5 s. 
     5) Chromatogram 
       FIG. 1  is a GC-MS chromatogram of extracts from the main root of woods-grown ginseng under the experimental conditions mentioned above. 
     Table 3 shows the characteristic chemical components (characteristic GC peaks) of extracts from the main root of woods-grown ginseng. 
     
       
         
               
             
               
               
             
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Essential Components of Extracts from 
               
               
                 Main Root of Woods-grown Ginseng 
               
             
          
           
               
                 Peak No 
                 Name 
               
               
                   
               
             
          
           
               
                 1 
                 5-Methyltetradecane 
               
               
                 2 
                 5-Isobutylnonane 
               
               
                 3 
                 5-Butylnonane 
               
               
                 4 
                 β-panasinsene 
               
               
                 5 
                 p-Acetylacetophenone 
               
               
                 6 
                 Paeonal 
               
               
                 7 
                 [1aR-(1aα,7α,7aα,7bα)]-1a,2,3,5,6,7,7a,7b- 
               
               
                   
                 Octahydro-1,1,7,7a-tetramethyl-1H- 
               
               
                   
                 cyclopropa[a] naphthalene 
               
               
                 8 
                 α-Caryophyllene 
               
               
                 9 
                 [1aR-(1aα.,4aα,7α,7aβ,7bα)]-Decahydro- 
               
               
                   
                 1,1,7-trimethyl-4-methylene-1H- 
               
               
                   
                 Cycloprop[e]azulene 
               
               
                 10 
                 Isocaryphyilene 
               
               
                 11 
                 8-Methylheptadecane 
               
               
                 12 
                 n-Elcosane 
               
               
                 13 
                 Crocetane 
               
               
                 14 
                 Falcarinol 
               
               
                 15 
                 cis,cis-Linoleic acid 
               
               
                   
               
             
          
         
       
     
       FIG. 2  is a GC-MS chromatogram of extracts from the main root of mountain cultivated ginseng. 
     Table 4 shows the characteristic chemical components (characteristic GC peaks) of extracts from the main root of mountain cultivated ginseng. 
     
       
         
               
             
               
               
             
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Essential Components of Extracts from the 
               
               
                 Main Root of Mountain-Cultivated Ginseng 
               
             
          
           
               
                 Peak No 
                 Name 
               
               
                   
               
             
          
           
               
                 1 
                 [1aR-(1aα,7α,7aα,7bα)]- 
               
               
                   
                 1a,2,3,5,6,7,7a,7b-Octahydro-1,1,7,7a- 
               
               
                   
                 tetramethyl-1H- 
               
               
                   
                 cyclopropa[a]naphthalene 
               
               
                 2 
                 Neoclovene 
               
               
                 3 
                 Decahydro-1,1,7-trimethyl-4- 
               
               
                   
                 methylene-1H- 
               
               
                   
                 cyclopropa[a]naphthalene 
               
               
                 4 
                 Isobutylnonane 
               
               
                 5 
                 Germacrene B 
               
               
                 6 
                 2,5-bis(1,1-dimethylethyl)-phenol 
               
               
                 7 
                 8-Methylheptadecane 
               
               
                 8 
                 n-Heptadecane 
               
               
                 9 
                 Falcarinolol 
               
               
                 10 
                 cis,cis-Linoleic acid 
               
               
                 11 
                 Tetratriacontane 
               
               
                 12 
                 n-Elcosane 
               
               
                   
               
             
          
         
       
     
     As shown in the figures, the chemical composition of the extracts of ginseng is very complicated, especially at high and low boiling points. The content of polyacetylene-alcohol (having a retention time of around 50 min) appears the highest. 
     5 batches of woods-grown ginseng were examined and the same number of batches of mountain-cultivated ginseng under the above-mentioned experimental conditions.  FIGS. 3 ,  4 ,  5  and  6  show GC-MS chromatograms of 3 batches of woods-grown ginseng and 2 batches of mountain cultivated ginseng 
     The above results manifest that extracts from different parts of the same ginseng vary considerably in quantity and content of characteristic chemical components. The differences are even greater than those between different varieties of ginsengs of the same part. The extracts from the same parts of similar samples are roughly the same in composition, proving the comparability between the extracts contents in the same part of the same variety. The comparison between the same parts in woods-grown ginseng and mountain cultivated ginseng can exclude the confounding factors of different parts, thus highlighting the difference of the samples per se. 
     A standard GC-MS fingerprint of the main root, the lateral root and rootlets from the woods-grown ginseng was established by analysis of the GC-MS chromatograms of the main root, lateral root and rootlets from 3 batches of woods-grown ginseng, which was obtained using the above described protocols as a mean and median. With the standard on hand, the resemblance was calculated between the 2 batches of woods-grown ginseng and 5 batches of mountain cultivated ginseng and the standard. The results were as follows: 
     1)The standard GC-MS fingerprint and similarity calculation of the extracts from main root of woods-grown ginseng. 
     1.1) Establishment of the standard of main root in woods-grown ginseng 
     Samples: sample 4, 5 and 6, or the woods-grown ginsengs of 11, 15 and 18 years old. 
     Preprocess of data: In the GC-MS chromatograms of three batches of woods-grown ginseng, 8 peaks were selected (marked numerically in  FIG. 7 ). The principle for selecting the peak was as follows: 
     1.1.1) The main peaks with a high response value were selected in that they were easy to identify and rectify. 
     1.1.2) The peaks selected appeared in each chromatogram, which enabled every peak to be used in comparisons. 
     1.1.3) The chromatographic peaks selected were distributed evenly in the chromatogram and the least square software method was employed in calculations, which made the adjacent chromatographic peak shift simultaneously. The selection ensured that all of the selected chromatographic peaks were used in calculations.  FIG. 8  is a chromatogram rectified by means of time calibration via the software of fingerprints of the sample, and  FIG. 9  is the standard. 
     
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Similarity in Groups of Main Roots of 
               
               
                 Three Batches of Woods-grown Ginseng 
               
             
          
           
               
                   
                   
                   
                 Related 
                 Related 
               
               
                   
                   
                 Sample 
                 Coefficient 
                 Coefficient 
               
               
                   
                 No 
                 No 
                 (median value) 
                 (mean value) 
               
               
                   
                   
               
               
                   
                 1 
                 4 
                 0.9405 
                 0.9359 
               
               
                   
                 2 
                 5 
                 0.9730 
                 0.9353 
               
               
                   
                 3 
                 6 
                 0.8768 
                 0.9441 
               
               
                   
                   
               
             
          
         
       
     
     1.2) Similarity Calculation 
     With the three woods-grown ginseng fingerprints established by the above method as standards, the similarity between the 3 batches of woods-grown ginseng, and 5 batches of mountain cultivated ginseng (see the GC-MS chromatograms in  FIG. 10 ) and the standards were calculated. (See results in Table 6). 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 Similarity between Different Samples of Mountain cultivated Ginseng 
               
               
                 and Standard of Main Roots of Woods-grown Ginseng 
               
             
          
           
               
                   
                   
                   
                 Related 
                 Related 
               
               
                   
                   
                   
                 Coefficient 
                 Coefficient 
               
               
                 Chromatogram 
                 Sample 
                   
                 (median 
                 (mean 
               
               
                 No. 
                 No. 
                 Sample 
                 value) 
                 value) 
               
               
                   
               
             
          
           
               
                 4 
                 1 
                 woods-grown 
                 0.9090 
                 0.7925 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (11-year-old) 
               
               
                 5 
                 2 
                 woods-grown 
                 0.8812 
                 0.7730 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (15-year-old) 
               
               
                 6 
                 3 
                 woods-grown 
                 0.9071 
                 0.8707 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (18-year-old) 
               
               
                 7 
                 7 
                 mountain 
                 0.4237 
                 0.3873 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (6-year-old) 
               
               
                 8 
                 8 
                 mountain 
                 0.4482 
                 0.4228 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (5-year-old) 
               
               
                 9 
                 9 
                 mountain 
                 0.4495 
                 0.3993 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (4-year-old) 
               
               
                 10 
                 10 
                 mountain 
                 0.3866 
                 0.3651 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (5-year-old) 
               
               
                 11 
                 11 
                 mountain 
                 0.5712 
                 0.5431 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (6-year-old) 
               
               
                   
               
             
          
         
       
     
     2. The standard GC-MS fingerprint and similarity calculation of extracts from lateral root of woods-grown ginseng. 
     2.1) Establishment of the standard of lateral root in woods-grown ginseng 
     Sample: samples 4, 5 and 6, or the woods-grown ginsengs of 11, 15 and 18 years old. 
     Preprocess of data: In the GC-MS chromatograms of three batches of woods-grown ginseng, 8 peaks were selected (marked numerically in  FIG. 11  to be used in calculations. See the rectified chromatogram in  FIG. 11 ; see the standard in  FIG. 12  and the similarity between the 3 batches of woods-grown ginseng in Table 7. 
     
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 Similarity in Groups of Lateral Roots of 
               
               
                 Three Batches of Woods-grown Ginseng 
               
             
          
           
               
                   
                   
                   
                 Related 
                 Related 
               
               
                   
                   
                 Sample 
                 Coefficient 
                 Coefficient 
               
               
                   
                 No 
                 No 
                 (median value) 
                 (mean value) 
               
               
                   
                   
               
               
                   
                 1 
                 4 
                 0.8973 
                 0.9515 
               
               
                   
                 2 
                 5 
                 0.9002 
                 0.9223 
               
               
                   
                 3 
                 6 
                 0.9511 
                 0.9520 
               
               
                   
                   
               
             
          
         
       
     
     2.2) Similarity Calculation 
     With the chosen standard of woods-grown ginseng established by the above method as criterion, the similarity of the GC-MS chromatograms of 2 batches of woods-grown ginseng and 5 batches of mountain cultivated ginseng was calculated ( FIG. 13 ). The result is as follows (Table 8): 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 8 
               
             
             
               
                   
               
               
                 Similarity between Different Samples of Mountain cultivated Ginseng 
               
               
                 and Standard of Lateral Roots of Woods-grown Ginseng 
               
             
          
           
               
                   
                   
                   
                 Related 
                 Related 
               
               
                 Chromatogram 
                 Sample 
                   
                 Coefficient 
                 Coefficient 
               
               
                 No. 
                 No. 
                 Sample 
                 (median value) 
                 (mean value) 
               
               
                   
               
             
          
           
               
                 1 
                 2 
                 Woods-grown 
                 0.7863 
                 0.7854 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (15-year-old) 
               
               
                 2 
                 3 
                 Woods-grown 
                 0.9510 
                 0.9555 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (18-year-old) 
               
               
                 3 
                 7 
                 Mountain- 
                 0.5739 
                 0.5349 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (6-year-old) 
               
               
                 4 
                 8 
                 Mountain- 
                 0.5719 
                 0.5202 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (5-year-old) 
               
               
                 5 
                 9 
                 Mountain- 
                 0.5537 
                 0.5842 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (4-year-old) 
               
               
                 6 
                 10 
                 Mountain- 
                 0.2449 
                 0.2441 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (5-year-old) 
               
               
                 7 
                 11 
                 Mountain- 
                 0.4441 
                 0.4846 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (6-year-old) 
               
               
                   
               
             
          
         
       
     
     3. The common GC-MS mode and similarity calculation of extracts from rootlets of woods-grown ginseng. 
     3.1) Establishment of the standard of rootlets of woods-grown ginseng 
     Sample: woods-grown ginseng of 11, 15 and 18 years old. 
     Preprocess of data: In the GC-MS chromatograms of three batches of woods-grown ginseng, 9 peaks were selected (marked numerically in  FIG. 14 ) to be used in calculations. See the rectified chromatograms in  FIG. 14 , the standard in  FIG. 15 , and the similarity within groups in Table 9. 
     
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 Similarity in Groups of Rootlets of 
               
               
                 Three Batches of Woods-grown Ginseng 
               
             
          
           
               
                   
                   
                   
                 Related 
                 Related 
               
               
                   
                   
                 Sample 
                 Coefficient 
                 Coefficient 
               
               
                   
                 No 
                 No 
                 (median value) 
                 (mean value) 
               
               
                   
                   
               
               
                   
                 1 
                 1 
                 0.9610 
                 0.9437 
               
               
                   
                 2 
                 2 
                 0.8937 
                 0.8948 
               
               
                   
                 3 
                 3 
                 0.8093 
                 0.9244 
               
               
                   
                   
               
             
          
         
       
     
     3.2) Similarity Calculation 
     With the woods-grown ginseng established by the above method as standard, the similarity of the GC-MS chromatograms of woods-grown ginseng and mountain cultivated ginseng was calculated ( FIG. 16 ). The result is as follows (Table 10): 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 10 
               
             
             
               
                   
               
               
                 Similarity between Different Samples of Mountain cultivated 
               
               
                 Ginseng and Standard of Rootlets of Woods-grown Ginseng 
               
             
          
           
               
                   
                   
                   
                 Related 
                 Related 
               
               
                   
                   
                   
                 Coefficient 
                 Coefficient 
               
               
                 Chromatogram 
                 Sample 
                   
                 (median 
                 (mean 
               
               
                 No. 
                 No. 
                 Sample 
                 value) 
                 value) 
               
               
                   
               
             
          
           
               
                 2 
                 5 
                 woods-grown 
                 0.9514 
                 0.9168 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (15-year-old) 
               
               
                 3 
                 6 
                 woods-grown 
                 0.8126 
                 0.9134 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (18-year-old) 
               
               
                 3 
                 7 
                 mountain 
                 0.6187 
                 0.7100 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (6-year-old) 
               
               
                 4 
                 8 
                 mountain 
                 0.6116 
                 0.7056 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (5-year-old) 
               
               
                 5 
                 9 
                 mountain 
                 0.6121 
                 0.6604 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (4-year-old) 
               
               
                 6 
                 10 
                 mountain 
                 0.2525 
                 0.2339 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (5-year-old) 
               
               
                 7 
                 11 
                 mountain 
                 0.6009 
                 0.6465 
               
               
                   
                   
                 cultivated 
               
               
                   
                   
                 ginseng 
               
               
                   
                   
                 (6-year-old) 
               
               
                   
               
             
          
         
       
     
     4. Conclusion 
     The samples of woods-grown ginseng and mountain cultivated ginseng can be well distinguished from each other by similarity calculation of GC-MS fingerprint of the main root, lateral root and rootlets. (Table 11 and Table 12) 
     
       
         
               
             
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 11 
               
             
             
               
                   
               
               
                 Similarity (median value) between Different Samples of 
               
               
                 Mountain cultivated Ginseng and Standard of 
               
               
                 Woods-grown Ginseng 
               
             
          
           
               
                   
                 Related coefficient (median value) 
               
             
          
           
               
                   
                 Sample 
                 Main root 
                 Lateral root 
                 Rootlets 
               
               
                   
                   
               
               
                   
                 Woods-grown 
                 0.88-0.91 
                 0.78-0.96 
                 0.81-0.96 
               
               
                   
                 ginseng 
               
               
                   
                 Mountain 
                 0.38-0.57 
                 0.24-0.57 
                 0.25-0.61 
               
               
                   
                 cultivated 
               
               
                   
                 ginseng 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 12 
               
             
             
               
                   
               
               
                 Similarity (median value) between Different Samples of 
               
               
                 Mountain cultivated Ginseng and Standard of 
               
               
                 Woods-grown Ginseng 
               
             
          
           
               
                   
                 Related coefficient (mean value) 
               
             
          
           
               
                   
                 Sample 
                 Lateral root 
                 Lateral root 
                 Rootlets 
               
               
                   
                   
               
               
                   
                 Woods-grown 
                 0.77-0.88 
                 0.78-0.96 
                 0.90-0.92 
               
               
                   
                 ginseng 
               
               
                   
                 Mountain 
                 0.36-0.54 
                 0.24-0.58 
                 0.24-0.71 
               
               
                   
                 cultivated 
               
               
                   
                 ginseng 
               
               
                   
                   
               
             
          
         
       
     
     Woods-grown ginseng and mountain cultivated ginseng can be distinguished from each other using the similarity coefficient calculation using standards based on GC-MS fingerprint of the main root, lateral root and rootlets. 
     Example 2 
     Identification of ginseng powder sample: sample: 2 samples of ginseng powder, marked as A and B respectively, were analyzed by gas chromatogram described as described above. Similarity calculation was conducted to the obtained GC-MS chromatograms. Because the main root in the ginseng sample generally accounts for 70-90% of the total weight of ginseng, the GC-MS chromatograms of the unknown sample and the GC-MS standard of main root in woods-grown ginseng were compared, and the result is as follows (normally, if identity of the part of ginseng from which extracts were made is known, extracts from main roots are compared to the main root standard, extracts from lateral roots are compared to the lateral root standard, and extracts from rootlets are compared to the rootlet standard): 
     
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 13 
               
             
             
               
                   
               
               
                 Similarity Calculation and Identification Results between Unknown 
               
               
                 Ginseng Samples and Standard of Woods-grown Ginseng (Main Root) 
               
             
          
           
               
                   
                 Related 
                 Related 
                   
               
               
                 Sample 
                 coefficient 
                 coefficient 
                 Identification 
               
               
                 No 
                 (median value) 
                 (mean value) 
                 Result 
               
               
                   
               
               
                 A 
                 0.7051 
                 0.7380 
                 woods-grown 
               
               
                   
                   
                   
                 ginseng 
               
               
                 B 
                 0.3871 
                 0.3568 
                 Mountain 
               
               
                   
                   
                   
                 cultivated 
               
               
                   
                   
                   
                 ginseng 
               
               
                   
               
             
          
         
       
     
     Result is as follows: A was woods-grown Ginseng, B was Mountain cultivated ginseng. 
     While particular embodiments of the invention have been shown and described, it will be obvious to those skilled in the art that changes and modifications may be made without departing from the invention in its broader aspects, and therefore, the aim in the appended claims is to cover all such changes and modifications as fall within the true spirit and scope of the invention.