To provide reference for quality control and authenticity identification of Arnebiae Radix medicinal materials and decoction pieces in the market. By studied on the ITS2 sequences’ characters of imported Arnebiae Radix，based on DNA barcoding and PCR-RFLP technologies.

The ITS2 region was selected as the DNA barcode sequence for comparison and identification of imported Arnebiae Radix and reference medicinal materials. The ITS2 sequences of imported Arnebiae Radix from different sources with reference medicinal materials were compared based on DNA barcoding and PCR-RFLP technologies.

After the restriction endonucliase AluI enzyme digestion，the agarose-gel electrophoresis results of 39 imported Arnebiae Radix samples showed that，only DH3 had bands at around 500 bp，and none bands between 100 bp and 300 bp. And the results of other imported Arnebiae Radix samples had two or three obvious bands between 100 bp and 300 bp. The ITS2 sequences of imported Arnebiae Radix samples were compared with the reference medicinal materials，among which DH3 had the largest differences of 15 bases compared to the reference medicinal materials，the ITS2 sequence of F2 was same to the reference medicinal materials，and other imported Arnebiae Radix samples had 1-9 bases difference compared to the reference medicinal materials. The clustering results showed that the imported Arnebiae Radix sample DH3 was clearly distinguished from other imported Arnebiae Radix samples and reference medicinal materials which was in a single branch. There were 14 samples，which were clustered together with the reference medicinal materials in one branch with support rate ≥50%.

The ITS2 region is selected to compare the similarities and differences of ITS2 sequences between imported Arnebiae Radix samples and reference medicinal materials based on DNA barcode and PCR-RFLP technologies，which provids a reference for effective identification of Arnebiae Radix medicinal materials and decoction pieces，and a strong guarantee for market supervision of Arnebiae Radix medicinal materials.

To design and screen specific primers for efficient amplification and identification of Arnebiae Radix from market based on the concept of nested PCR.

Nested primers was designed using the software of Primer Premier 5 based on the ITS sequence of Arnebia euchroma and the ITS2 sequence of non-pharmacopoeial Arnebiae Radix. The amplification efficiency of genomic DNA by ITS2 universal primers PCR and nested PCR was compared. The genomic DNA of Arnebiae Radix was amplified directly by nested primers and was detected by agarose gel electrophoresis. The specific primers designed for Arnebiae Radix based on the fragment length and variation sites’ coverage of the amplified product was evaluated.

A total of 11 primers were selected for synthesis after the primers were designed by Primer Premier 5 software. The amplification efficiency of nested PCR was superior to ITS2 universal primers PCR in genomic DNA of Arnebiae Radix. The results of nested primers directly amplified genomic DNA of Arnebiae Radix by agarose gel electrophoresis were better than those of ITS2 primers，and showed a single band. Four pairs of primers，AE-9S/AE-2A，AE-4S/AE-10A，AE-12S/10A，AE-29S/AE-29A，were determined to be suitable for the identification of Arnebiae Radix.

On the basis of DNA barcode identification and nested PCR technology，4 pairs of specific primers are identified which can be used to effectively distinguish Arnebia euchroma from the mainstreamed non-pharmacopoeial Arnebiae Radix in the medicinal materials market，providing reference for the subsequent research and development of identification methods for Arnebiae Radix and other traditional Chinese medicines.

To determine the color of Arnebiae Radix, and the contents of six main purple pigment components (acetylshikonin，β-acetoxyisovalerylalkannin, deoxyshikonin, isobutylshikonin，β，β’-dimethylacrylalkannin and isovalerylshikonin) in Arnebiae Radix, and to study on the correlation between the color of Arnebiae Radix and the contents of six main purple pigment components.

The L，a，and b values of the sample powder were determined using a spectrophotometer to characterize the color of Arnebiae Radix. The International Commission on Illumination (CIE) had developed a Lab color model，which was a digital description of human vision. A higher L value indicated greater brightness，a higher a value indicated redness and a lower a value indicates greenness，and a higher b value indicated yellowing and a lower b value indicates blueness. The contents of purple pigment components were determined using high performance liquid chromatography (HPLC)，and the correlation between L，a，and b values and the contents of six main purple pigments was calculated using SPSS software.

The contents of acetylshikonin in 135 batches of samples ranged from 0.01% to 3.39%. The contents of β-acetoxyisovalerylalkannin ranged from 0.00% to 1.95%. The contents of deoxyshikonin ranged from 0.00% to 0.23%. The contents of isobutylshikonin ranged from 0.01% to 1.13%. And the contents of isovalerylshikonin ranged from 0.02% to 2.88%. The contents of β，β’-dimethylacrylalkannin ranged from 0.01% to 2.17%. There was a significant negative correlation between the contents of acetylshikonin，β-acetoxyisovalerylalkannin and isobutylshikonin and the L (black_white) chromaticity value of Arnebiae Radix，with a Spearman correlation coefficients between -0.138 and -0.222. The chromaticity value of a (red_green) was related to the five components other than acetylshikonin，which were β-acetoxyisovalerylalkannin，deoxyshikonin，isobutylshikonin，β，β’-dimethylacrylalkannin，and isovalerylshikonin，with a spearman correlation coefficients between 0.176 and 0.355；b (blue_yellow) chromaticity value was related to the five components other than β-acetoxyisovalerylalkannin，which were acetylshikonin，deoxyshikonin，and isobutylshikonin，β，β’-dimethylacrylalkannin. β，β’-dimethylacrylalkannin was positively correlated with a coefficient of 0.290，and negatively correlated with the other four components with a coefficients between -0.325 and -0.633.

It is recommended that the assay limits of Arnebiae Radix be revised to β，β’-dimethylacrylalkannin not less than 0.30% and isovalerylshikonin not less than 0.29%.

To compare the regulatory effects of authentic and counterfeit Arnebiae Radix on intestinal flora in mice based on metagenomic sequencing.

Firstly，24 clean grade female BLAB/C mice were randomly divided into 3 groups：blank control group，A1 (Arnebia euchroma) group and A2 (Arnebiae Radix whose origins were not included in Chinese Pharmacopoeia) group. After gavage arrived at the specified time，colon contents (feces)，ileal contents (feces) and small intestinal contents (feces，except ileal parts) were extracted for intestinal flora analysis. Genomic DNA was extracted and amplified by PCR from the extracted mouse intestinal contents. The PCR products were mixed and purified. Then library was constructed and sequenced. After quality control of sequencing data and removal of chimera sequence，the final effective data was obtained. Operational taxonomic unit(OTU) clustering and species annotation were performed on the obtained valid data，and sample diversity analysis was conducted.

In this study，both A1 (Arnebia euchroma) group and A2 (Arnebiae Radix non-pharmacopoeia) group reduced the diversity of mice colon microbiota. At the phylum level，group A1 significantly increased the abundance of Firmicutes in the small intestine and ileum，and groups A1 and A2 significantly increased the relative abundance of Bacteroidetes in the colon. At the genus level，group A1 significantly increased the relative abundance of Lactobacillus in the small intestine of mice，and group A2 significantly increased the relative abundance of Lactobacillus in the ileum of mice. Group A1 increased the relative abundance of Lactobacillus，one of the dominant bacteria in the colon of mice，and group A2 increased the relative abundance of Bacteroides. Alistipes mainly existed in the colon，A2 group significantly reduced the relative abundance of Alistipes in the colon of mice，while Alistipes in the A1 group was cultivated.

According to the results of the regulation effect of intestinal flora，the intestinal flora regulation effect in the two experimental groups of the authentic Arnebiae Radix and its confusion products from markets are not consistent，the results of this study can provide a theoretical basis for further exploring the mechanism of Arnebiae Radix.

To compare the quality of wild and cultivated Arnebiae Radix，using macroscopic investigation and chemometric analysis of the different components in wild and cultivated Arnebiae Radix from three different habitats.

Wild and cultivated Arnebiae Radix were collected and their macroscopic features were compared. Using the ACQUITY UPLC BEH C₁₈ (2.1 mm×100 mm，1.7 μm) column，with acetonitrile-0.05% formic acid water as the mobile phase，the contents of D-shikonin，acetylshikonin，β-acetoxyisovalerylshikonin，isobutyrylshikonin，β，β’-dimethylacrylalkannin and isovalerylshikonin in 48 batches of wild and cultivated Arnebiae Radix were determined. The detection wavelength was 275 nm and the flow rate was 0.2 mL·min^-1. PCA and OPLS-DA were performed to reveal the differential components of wild and cultivated Arnebiae Radix.

There were great differences in macroscopic features of wild and cultivated Arnebiae Radix，and the linear relationship between the contents of six naphthoquinone components was good. The correlation coefficients were above 0.999，the average recovery rates were 93.4%-102.9%，and the RSDs were less than 3.0%. The contents of six components in different batches of wild and cultivated Arnebiae Radix were quite different，and the contents of D-shikonin and acetylshikonin in wild products were significantly higher than those in cultivated products，indicating that there were still certain differences between wild products and cultivated products. The PCA model established could distinguish wild products and cultivars，and two differentiating components in wild products and cultivars were revealed by OPLS-DA，namely isobutyryl shikonin，β，β’-dimethylacrylalkannin.

By comparing the core size，cork curl degree and specific odor of wild and cultivated products，the two can be identified. The established content determination method is repeatable，specific，stable and feasible. The differential components in wild and cultivated Arnebiae Radix in three different regions are identified，which provides a basis for the quality control of Arnebiae Radix and provides ideas for expanding the source of Arnebiae Radix.

To establish a method for simultaneous determination of neochlorogenic acid，chlorogenic acid，cryptochlorogenic acid，cynarin，galuteolin，isochlorogenic acid B，isochlorogenic acid A，isochlorogenic acid C in Shanjujiangya capsules by HPLC and analyze compositional change of Chrysanthemi Flos combined with law of quantity transfer．

The analysis was performed on Waters Symmetry C₁₈ column (250 mm×4.6 mm，5 μm)，with mobile phase composed of acetonitrile -0.1% phosphoric acid solution at a flow rate of 1.0 mL·min^-1 in gradient elution mode. The column temperature was 30 ℃ and the detection wavelength was 328 nm. The transfer rates of the above eight components were used as the indexes for quality evaluation to study the quantity value of transfer rule from the decoction piece to the extracting solution.

The results showed that the determination of eight components manifested a good linear relationship in the range of mass concentration (r>0.999 9)，the average recoveries of neochlorogenic acid，chlorogenic acid，cryptochlorogenic acid，cynarin，galuteolin，isochlorogenic acid B，isochlorogenic acid A，isochlorogenic acid C were 98.3%-101.9%，with RSDs of 0.066%-0.64%. The contents of the above 8 components in 3 samples were 0.257-0.279 mg·g^-1，0.629-0.650 mg·g^-1，0.402-0.476 mg·g^-1，0.454-0.539 mg·g^-1，1.118-1.278 mg·g^-1，0.653-0.740 mg·g^-1，0.659-0.706，1.138-1.167 mg·g^-1，respectively.

The HPLC method established in this study is simple，repeatable and stable. The analysis of quantity transfer provides data support for the establishment of content methods and the formulation of limits. This study can provide basis for quality control method of Shanjujiangya capsules.

To establish a quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of 7 flavanoids (5-hydroxy-6，7，3’，4’-tetramethoxyflavone，apigenin，hispidulin，kaempferol，jaceosidin，eupatilin and casticin) in Artemisiae Argyi Folium.

The HPLC system consisted of the Agilent ZORBAX SB-C₁₈ column (150 mm×4.6 mm，5 μm) column with gradient elution of acetonitrile and 0.2% phosphoric acid as the mobile phase at a flow rate of 1.0 mL·min^-1，a detection wavelength of 350 nm，and a column temperature of 30 ℃. Eupatilin was selected as the internal reference substance，the relative correction factors between eupatilin and the other 6 flavanoids were established，and the contents of these 7 constituents in samples were calculated to realize QAMS. At the same time，compared with the external standard method to verify the accuracy and feasibility of the QAMS method.

Within a certain linear range，the relative correction factors between eupatilin and 5-hydroxy-6，7，3’，4’-tetramethoxyflavone，apigenin，hispidulin，kaempferol，jaceosidin as well as casticin were 0.958，1.387，1.000，0.950，0.957 and 1.297，respectively (RSDs of RCFs were less than 2.0%). The contents of 5-hydroxy-6，7，3’，4’-tetramethoxyflavone，apigenin，hispidulin，kaempferol，jaceosidin，eupatilin，casticin in 20 batches of Artemisiae Argyi Folium were 0.031 4-0.623 5 mg·g^-1，0.000 9-0.092 6 mg·g^-1，0.020 6-0.170 7 mg·g^-1，0.011 0-0.184 7 mg·g^-1，0.011 7-0.864 0 mg·g^-1，0.253 2-2.555 0 mg·g^-1 and 0.015 6-0.250 7 mg·g^-1，respectively.

Using eupatilin as the internal reference，QAMS method for 7 flavanoids is established. The method is accurate and reliable，and can be used for quality control and quantitative analysis of Artemisiae Argyi Folium.

To establish HPLC fingerprint and determine caffeic acid，isoquercitrin，quercitrin，rosmarinic acid，lithospermic acid and salvianolic acid B in Tournefortia sibirica Linnaeus，and to provide evidence for quality control of Tournefortia sibirica Linnaeus.

The chromatographic separation was performed on an Agilent Eclipse Plus C₁₈ column (250 mm×4.6 mm，5 μm) with gradient elution (0-60 min，9%B→38%B) of 0.2% phosphoric acid(A) and acetonitrile(B) ． The detection wavelength was 330 nm. The column temperature was kept at 40 ℃ and the flow rate was 1.0 mL·min^-1. The HPLC fingerprint of all batches of Tournefortia sibirica Linnaeus was established using Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 edition) and the common peaks were identified by reference standards. Six constituents in Tournefortia sibirica Linnaeus were quantified.

Eleven common peaks were confirmed，and 6 common peaks were identified by reference standards including caffeic acid，isoquercetin，quercetin，rosmarinic acid，purple oxalic acid，and salvianolic acid B. The similarities of 17 batches samples were 0.931 to 0.996. By the methodology examination，RSDs for the precision test were 1.9%，1.0%，1.4%，0.19%，1.1% and 0.32%，respectively. RSDs for the reproducible test were 3.5%，2.3%，3.3%，0.14%，1.1% and 0.19%，respectively. RSDs for the stability test were 0.80%，1.1%，1.7%，0.52%，0.54% and 0.78%，respectively. Caffeic acid，isoquercitrin，quercitrin，rosmarinic acid，lithospermic acid and salvianolic acid B had good separation and showed good linearity in their respective linear ranges. The average recoveries ranged from 95% to 105%，and the contents (calculated with reference to the dried drug) were between 0.004% to 0.013%，0.030% to 0.259%，0.032% to 0.256%，0.256% to 1.246%，0.018% to 0.072% and 0.062% 0.499%，respectively.

The established HPLC fingerprint and quantification method is stable and reliable，which can provide basis for the quality control of Tournefortia sibirica Linnaeus.

To establish an HPLC-MS/MS method for simultaneous determination of 11 components（harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone，harpagoside，vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ，and atractylenolide Ⅰ）in Gengnianning.

HPLC assay was performed on C₁₈ column（100 mm×2.1 mm，1.9 μm）with a mixture of methanol and 0.1% formic acid as the mobile phase in gradient elution at a flow rate of 0.3 mL·min^-1. The column temperature was 25 ℃ and the injection volume was 1 μL. Detection was carried out on a triple quadrupole mass spectrometer in positive ion mode（harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone and harpagoside）and negative ion mode（vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ，and atractylenolide Ⅰ）using an electrospray ion source（ESI）. Multiple reaction monitoring（MRM）mode was employed.

The calibration curves were linear within the ranges of 1.485-29.71 μg·mL^-1，1.620-32.40 μg·mL^-1，7.801-156.0 μg·mL^-1，0.518-10.35 μg·mL^-1，0.167-3.333 μg·mL^-1，0.359-7.179 μg·mL^-1，1.455-29.10 μg·mL^-1，1.520-30.40 μg·mL^-1，0.160-3.205 μg·mL^-1，0.143-2.864 μg·mL^-1 and 0.157-3.136 μg·mL^-1 for harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone，harpagoside，vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ，and atractylenolide Ⅰ，respectively. All 11 components showed good linearity（r≥0.998 0）. The average recoveries（n=6）were in the range of 95.9%-102.6% with RSDs within 0.90%-3.0%. The contents of harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone，harpagoside，vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ and atractylenolide Ⅰ in 10 tested samples from 5 manufactures were 14.8-104.5，37.6-288.5，335.8-1 332.8，6.2-10.1，6.6-61.8，13.7-75.1，57.4-132.8，16.9-70.6，11.8-33.9，3.4-15.4 and 6.5-12.9 μg·g^-1.

The developed method is accurate and sensitive. It can be used in quality control of Gengnianning.

To establish a dual-wavelength method for the determination of amylose and amylopectin in potato starch in Tibet.

The determination wavelengths of amylopectin and amylopectin in Tibetan potato were 623 and 557 nm，respectively，and the reference wavelengths were 498 and 729 nm. The contents in 11 batches of starch samples collected were determined and analyzed.

The results showed that the mass concentration of amylose and amylopectin showed a good linear relationship (r=0.999 9，r=0.999 8) in the range of 0- 66 μg·mL^-1 and 0-90 μg·mL^-1，respectively. The average recoveries of amylose and amylopectin were 102.8% (RSD=1.8%) and 98.5% (RSD=2.2%). The precision (RSD=0.071%，RSD=0.31%)，repeatability (RSD=0.26%，RSD=2.4%) and stability (RSD=0.14%，RSD=1.4%) were all in compliance with regulations. The contents of amylose and amylopectin in Tibetan potato starch were 36.74% and 45.87%，respectively. Compared with the commercially available potato starch，the amylose content of Tibetan potato starch was significantly increased (P<0.001)，while the amylopectin content was significantly decreased (P<0.05).

The method is proved to be suitable for the quality evaluation of potato starch in Tibet. There are significant differences in the contents of amylopectin and amylopectin between potato starch in Tibet and potato starch in the market，which provides an experimental basis for the establishment of quality standards of potato starch in Tibet and the development of high-value products.