| BaiZhi is the dried root of Umbelliferae Angelica dahurica (Fisch. ex Hoffm.) Benth. et Hook. f. or Angelica dahurica (Fisch. ex Hoffm.) Benth. et Hook. f. var. formosana (Boiss) Shan et Yuan, it’s a commonly used Chinese medicine. BaiZhi is mainly produced in Sichuan, Zhejiang, Hebei, Henan and other provinces, where it is ChuanBaiZhi, HangBaiZhi, QiBaiZhi, YuBaiZhi. In the course of standardized planting ChuanBaiZhi, seed production of ChuanBaiZhi is seriously lagging behind. There is evaluating the quality of ChuanBaiZhi seed full using of the experience of local farmer, having the common phenomenon of their own breeding, serous lacking its legal standards, what is extremely harmful to the production of quality ChuanBaiZhi. In the study of quality standards of ChuanBaiZhi seed, combining the literature, we found large differences in the quality of seeds coming from different regions, and the phenomenon that GongBaiZhi seed instead of Angelica seed. So, we should improve standards of Angelica seeds and identificate different regions of Angelica seeds at the same time.In the article, the test method of ChuanBaiZhi seed was studied, the grading standards of ChuanBaiZhi seed were developed, the grading standards by field testing were proved. They provided standardized criteria for production, circulation, restriction, using of ChuanBaiZhi seed, and a reference for other Chinese herbal medicines. We tentatively identified the difference in Angelica seeds including ChuanBaiZhi, GongBaiZhi, HangBaiZhi, QiBaiZhi,YuBaiZhi, by Morphology and histology, Chemical Composition, Proteins,etc, what gave theoretical and technical support to identify Angelica seeds from different regions. The results were as foliows:1 The study on quality standards of ChuanBaiZhi seed1.1 The study on the test methods of ChuanBaiZhi seedReference to the international inspection of seed, The inspection of crop seeds (GB/T3543-1995) and the assessment objectives and research of Chinese herbal seeds standard platform, we systematically conducted a methodological study from seeds samping, clarity analysis, germinal;on lest, identification of authenticity, moisture, viability, weight, health, using two batches of ChuanBaiZhi seed from GAP base. We developed ChuanBaiZhi seed testing standards: Minimum sampling amount was 12g; clarity analysis after 20 mesh sifting: morphhological identification and size measurement; 1000-grain weight; putting seeds on the sand, illuminantion 12 hour, temperalure of 25℃/18℃ (day/nighi). coant time 9～30d; seeds moisture determination was drying 4 hour in lower the thermostat(105±2)℃, seeds viability determination was staining by 0.7%TTC; seeds heallhy determination was direet inoculating on PDA medium, counting after 28℃, culture 3 days. In the study, the lest method of ChuanBaiZhi seed was first studied and developed, what gave the digital support to development of inspection procedures and grading standards of ChuanBaiZhi seed.1.2 The study on the grading standards of ChuanBaiZhi seedIn the study, several indicators o[" ChuanBaiZhi seed were first measured, and the grading standards of ChuanBaiZhi seed were deveh-ped, which were considered Io be scientific and applicable. They gave the theoretical support to quality contral and rational use of ChuanBaiZhi seed. We determined the indicators of germination rate, grain weight, viability, clarity and moisture of 56 batches of ChuanBaiZhi seeds, using the lest methods of ChuanBaiZhi seed, and standardized the research data by grade math principles about K-means clustering analysis, and developed the grading standards of ChuanBaiZhi seed whose main indicaiors include germination rate and grain weight, and whose reference indicators including viability, clarity and moisture. The grading standards of ChuanBaiZhi seed were as follows: Ⅰ grade seed germination rate≥54.6%, grain weight≥3.48g, viability≥69.2%, clarity 91.8%, moisture≤10%;Ⅱ grade seed germination rate 38.6%～ 54.6%, grain weight 3.06g～3.48g, viability≥55.7% ～69.2%, clariiy≥ 84.6%～91.8%, moisture≤10%; Ⅲ grade seed germination rate 28.0%～38.6%, grain weight 2.70g～3.06g, viability 45.0%～55.7%, clarity 75.0%～84.6%, moisture≤10%.1.3 The study on correlation between the grading of ChuanBaiZhi seed and the indicators of growth herbs, medicinal yiehl and qualityIn the study, the correlation between the grading of ChuanBaiZhi seed and the indicators of growth herbs, medicinal yield and quality was first studied. The variation of the grading of ChuanBaiZhi seed and plant growth, development and yield and quality of medicines were found out. The grading standards of ChuanBaiZhi seed were considered to be scientific and applicable, and they gave the technical support to the production of high quality ChuanBaiZhi medicine.Based on the grading standards of ChuanBaiZhi seed what are obtained in previous studies, The ChuanBaiZhi seeds were divided into three grades:I grade, Ⅱ grade and Ⅲ grade. Using GongBaiZhi seed and different of ChuanBaiZhi seed, we continuously measured and evaluated a lot of indicators including agronomic traits, fresh and dry weight of various vegetative organs, and we measured the yield, content of volatile oil and coumarn, in order to determine whether the grading standards of ChuanBaiZhi seed were reasonable. The experimental results were as follows:Ⅰ grade seed and Ⅱ grade seed are better than Ⅲ grade seed and GongBaiZhi seed in growth and development of plants. The higher the grading, the higher the yield, and GongBaiZhi seed yielded too low, was not suitable for production. The higher the grading, the higher the content of oxypeucedanin and imperatorin in medicinal material. Different levels had little impact on the content of volatile oil.2 The study on identification of Angelica seeds from different regions2.1 The comparative study on Morphology and histology about Angelica seeds from different regionsIn the study, the comparative study on morphology and histology about Angelica seeds from different regions was first conducted, and the differences in morphology and histology about Angelica seeds from different regions were found out, which provided the morphology and histology basis for identification of Angelica seeds.We did character identification and microscopic identification, including schizocarps shape, size, color, surface characteristics, the distribution of the cross-section tubing, and the related indicators of cross-section tubing. The experimental results were as follows:There was little difference in schizocarps morphology from different regions. HangBaiZhi schizocarps and ChuanBaiZhi schizocarps were longer, QiBaiZhi schizocarps were shorter; ChuanBaiZhi schizocarps and YuBaiZhi schizocarps were wider, HangBaiZhi schizocarps and BoBaiZhi schizocarps were narrower; The proportion of ChuanBaiZhi and YuBaiZhi seed wing was larger, the proportion of HangBaiZhi and BoBaiZhi seed wing was smaller. There were large differences in sehizocarps cross-section tubing from different regions. The perimeter of each tubing of HangBaiZhi schizocarps and YuBaiZhi schizocarps was longer, the area of each tubing of HangBaiZhi schizocarps and YuBaiZhi schizocarps was largr, and each tubing of HangBaiZhi schizocarps and YuBaiZhi schizocarps was more full; The perimeter and area of each tubing of ChuanBaiZhi schizocarps was general, and each tubing of ChuanBaiZhi schizocarps was full; The perimeter of each tubing of GongBaiZhi schizocarps was general, the area of each tubing of GongBaiZhi schizocarps was smaller, and each tubing of GongBaiZhi schizocarps was compare shriveled; The perimeter of each tubing of BoBaiZhi schizocarps was shorter, the area of each tubing of BoBaiZhi schizocarps was smaller, and each tubing of BoBaiZhi schizocarps was full; The perimeter of back dough tubings tubing of QiBaiZhi schizocarps was shorter, the area of back dough tubings of QiBaiZhi schizocarps was smaller, and back dough tubings of QiBaiZhi schizocarps were full, the perimeter of hopson surface tubings tubing of QiBaiZhi schizocarps was longer, the area of hopson surface tubings of QiBaiZhi schizocarps was larger, and hopson surface tubings of QiBaiZhi schizocarps were full.2.2 The comparative study on chemical composition about Angelica seeds from different regionsIn the study, the coumarin HPCL fingerprint and the volatile oils GC fingerprint of Angelica seeds from different regions were first studied, and the differences in coumarin and volatile oils about Angelica seeds from different regions were found out, which provided the chemical composition basis for identification of Angelica seeds.2.2.1 The study on the coumarin HPCL fingerprint of Angelica seeds from different regionsWe established the coumarin HPCL fingerprint of Angelica seeds from different regions, and got 10 common peaks in the control of chromatography after correction. The coumarin HPCL fingerprint of Angelica seeds from different regions was good, there was a large difference in the peak area of common peaks because of different origins. By the cluster analysis, Angelica seeds from different regions were divided into five categories:ChuanBaiZhi seeds, GongBaiZhi seeds, HangBaiZhi seeds, YuBaiZhi seeds, QiBaiZhi seeds.2.2.2 The study on volatile oils GC fingerprint of Angelica seeds from different regionsWe established the volatile oils GC fingerprint of Angelica seeds from different regions, and got 27 common peaks in the control of chromatography after correction. The volatile oils GC fingerprint of Angelica seeds from different regions was good, there was a large difference in the peak area of common peaks because of different origins. By the cluster analysis, Angelica seeds from different regions were divided into four categories:GongBaiZhi seeds, YuBaiZhi seeds, QiBaiZhi seeds, ChuanBaiZhi seeds and HangBaiZhi seeds.The study results were as follows:there were the olefin, the alkanes, the alcohol, the ester and the ketone in the volatile oils of Angelica seeds from different regions. 84 kinds of compounds were identified, and 15 kinds of common compounds were identified.57 kinds of compounds were identified in ChuanBaiZhi seeds(SN2), 54.67% in the total content, olny 3 kinds of compounds in ChuanBaiZhi seed.32 kinds of compounds were identified in GongBaiZhi seeds(SNG2),68.99% in the total content, olny 2 kinds of compounds in GongBaiZhi seed.58 kinds of compounds were identified in HangBaiZhi seeds(PA4),66.26% in the total content, olny 8 kinds of compounds in HangBaiZhi seed.49 kinds of compounds were identified in QiBaiZhi seeds(AG2),46.66% in the total content, olny 8 kinds of compounds in QiBaiZhi seed.50 kinds of compounds were identified in YuBaiZhi seeds(YZ1), 64.78% in the total content, olny 2 kinds of compounds in YuBaiZhi seed. The main component of volatile oil comparative analysis showed that there was identification of ChuanBaiZhi seed and GongBaiZhi seed by 1,2,3,4,4a,5,6,8a-octahydro-7-methyi-4-methylene-i-(1-methylethyl)-, (1.alpha.,4a.alpha.,8a.alpha.)-Naphthalene and its relative content, and that there was identification of Angelica seeds from different regions by Bicyclo[4.1.0]hept-2-ene, 3,7,7-trimethyl-,1,2,4-Metheno-1H-indene,octahydro-1,7a-dimethyl-5-(1-methylethyl)-,[1S-(1.alpha.,2.alpha.,3a.beta.,4.alpha.,5.alpha.,7a.beta.,8.beta.)]- and seychellene and its relative content.2.3 The comparative study on proteins about Angelica seeds from different regionsIn the study, the study on the proteins SDS-PAGE fingerprint of Angelica seeds from different regions was first done, and the differences in the proteins about Angelica seeds from different regions were found out. The preliminary study on proteins Thz-TDS of Angelica seeds from different regions was first conducted, and the differences in the proteins Thz absorption of proteins about Angelica seeds from different regions were found out. They provided the protein basis for identification of Angelica seeds.2.3.1 The study on the proteins SDS-PAGE fingerprint of Angelica seeds from different regionsWe established the proteins SDS-PAGE fingerprint of Angelica seeds from different regions. ChuanBaiZhi seed had 13 standard characteristic protein bands, GongBaiZhi seed had 11 standard characteristic protein bands, HangBaiZhi seed had 17 standard characteristic protein bands, QiBaiZhi seed had 16 standard characteristic protein bands, YuBaiZhi seed had 16 standard characteristic protein bands. All the standard characteristic protein bands were quantified relativized. By the principal component analysis and the cluster analysis, Angelica seeds from different regions were divided into five categories:ChuanBaiZhi seeds, GongBaiZhi seeds, YuBaiZhi seeds, QiBaiZhi seeds, HangBaiZhi seeds.2.3.2 The preliminary study on proteins Terahertz time-domain spectroscopy (Thz-TDS) of Angelica seeds from different regionsThe THz absorption spectrums of Angelica seeds from different regions mainly lay on Thz low frequency,including 0.17THz,0.18 THz,0.22 THz,0.27 THz and 0.28 THz. Compared with other seeds, GongBaiZhi seed had characteristic absorption in the 0.29 THz, which could be used to identify other seeds. QiBaiZhi seed had characteristic absorption in the 0.12 THz,0.14 THz and 0.25 THz. The THz absorption had substantially identical position of ChuanBaiZhi seeds and HangBaiZhi seeds, but the absorption coefficient of HangBaiZhi seeds was greater than the absorption coefficient of ChuanBaiZhi seeds. |
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