Study On Fingerprints Of Processed Largehead Atractylodes Hizome

BackgroundFingerprints can fully reflected overall chemical characteristics and overall quality of traditional Chinese medicine. Fingerprints can also overcome the disadvantages of previous quality control methods which only focus on single component and is a more reasonable quality control model. State Food and Drug Administration in 2000 enacted the "Technical requirements on fingerprints researched of traditional Chinese medicine injections (temporary)" to regulate the fingerprint of traditional Chinese medicine research which promoted the fingerprint researches and the unique advantages of fingerprints have been noticed gradually. For more effective quality control of different processed Largehead Atractylodes Rhizome and revealing the connotation of effect changing after processing, HPLC method and GC-MS method were established and analyzed in this paper.ObjectiveFingerprints of different processed Largehead Atractylodes Rhizome were established by HPLC and GC-MS. The differences and similarity were compared for the standard processing technology, quality control, and further developing. Methods1. Development of HPLC fingerprint of different processed Largehead Atractylodes RhizomeChromatographic conditions:Kromasil C18 (250mm×4.6mm,5μm) column and diode array detector (DAD) were used. A gradient solvent program consisted with acetonitrile (A) and water (B) was used as mobile phase in gradient elution. The column was eluted at the flow-rate at 1mL·min-1, column temperature was 25℃and detection wavelength was 242nm; The injection volume was 10μL. Under these conditions, HPLC fingerprints of different processed materials were determined and changes among processed materials were compared.2. Development of GC-MS Fingerprint of volatile oil different processed productsChromatographic conditions:The DB-5MS capillary column (30m×250μm×0.25μm) was used and the injection volume was 1.0μl; Split injection (5:1) was adopted at 250℃; He was used as carrier gas at constant flow-rate of 1ml·min-1, and the average line speed was 37cm/sec. The temperature program was as follows: initial temperature 100℃increasing to 130℃at the interval of 3℃·min-1, keeping 10min; then increasing to 250℃at the interval of 6℃·min-1. The interface temperature was 280℃.MS conditions:ion source temperature was 230℃, quadrupole temperature was 150℃, ionization voltage was 1906V, ionization energy was 70eV, mass scan range was 30-450amu. NIST05, WILEY275 database were adopted.Different processed materials were prepared by fried crude Largehead Atractylodes Rhizome with bran, soil or deeply fried.Volatile oil was extracted by steam distillation to extract from the fried products. GC-MS fingerprints were established with above chromatographic conditions and changes in. Results1. Determination of HPLC fingerprints of different processed materials HPLC fingerprints of 10 batches Largehead Atractylodes Rhizome, bran-fried Largehead Atractylodes Rhizome, soil-fried Largehead Atractylodes Rhizome, deeply fried Largehead Atractylodes Rhizome and fried Largehead Atractylodes Rhizome were determined and 17 common peaks were demarcated. The peaks of different processed products were similar with the similarity exceeding 0.9. The model of common peaks was developed and it was found that common peaks existed in different processed materials and their retention time and relative retention time were accordant but intensity varied. It was indicated that The difference of common peak area between soil-fried Largehead Atractylodes Rhizome and deeply fried material was obvious, nevertheless the difference of common peak area between bran-fried Largehead Atractylodes Rhizome and fried material was small.2. Determination of GC-MS fingerprints of essential oil of different processed Largehead Atractylodes Rhizomey-caryophyllene,β-caryophyllene,γ-elemene,α-humulene,β-eudesmene,α-gurjunene,δ-himachalene, germacrene B,4,5-dehydrogenation Isolongifolene,β-Citrus aurantium L. cv. Zhulan, Atractylon, a-eudesmol, juniper camphor, 6,7-dihydro-methoxy furan coumarin factors Selina-3,7(11)-diene, oleic acid were common components of 10 batches processed Largehead Atractylodes Rhizome, these components were about 85.5% to 98.2% of total volatile oil, the relative peak areas of different processed products were different, the peak areas and retention time were the characteristics of fingerprints.ConclusionsHPLC fingerprint of different processed products and GC-MS fingerprints of volatile oil can be used for evaluating the composition differences between Largehead Atractylodes Rhizome and its processed material. The analysis of component changes in different processed materials can also be used as a quality control method. It was found that increased trends of atractylenolideⅠandⅢcontents and decreased trend of atractylone may be the reason for the increasing effects of processed Largehead Atractylodes Rhizome.