To establish a hydrogen nuclear magnetic quantitative method which can rapidly determine the contents of linalool and protocatechuate in Homalomena occulta.

Using dimethyl sulfoxide-d₆（DMSO-d₆）as the test solvent and pyrazine -as the internal standard, ¹H-q NMR measurement was performed on a 400 MHz spectrometer. The quantitative resonance peaks of pyrazine, Linalool and protocatechuate were δ 8.66, δ 5.90 and δ 8.60, respectively.

The linear correlation coefficients of linalool and protocatechuate were good with r of 0.999 7 and 0.998 6, respectively. The RSDs of precision were 0.30% and 0.40%, and the recoveries were 98.8%-100.7%（RSD=3.7%） and 99.4%-100.6%（RSD=4.4%）, respectively, which indicated that the established method was accurate, stable and feasible. And the whole detection process was completed in about three minutes. The contents of 2 components in 4 batches of Homalomena occulta were 0.328-0.398 mg·g^-1（linalool）and 0.559-0.630 mg·g^-1 （protocatechuate）, respectively.

The ¹H-q NMR method has the advantages of simple operation, fast analysis speed and specificity, and can be used for the simultaneous determination of two active components in Homalomena occulta, providing a scientific basis for the overall quality evaluation and quality control of Homalomena occulta.

To determine the content of acrylamide in “8 famous medicines from Zhejiang”, and to carry out the preliminary risk assessment based on the determination results.

After optimization of QuEChERS extraction, the content of acrylamide was determined by the method of isotope-labeled internal standard-ultra high performance liquid chromatography-tandem mass spectrometry (isotope-labeled IS-UHPLC-MS/MS). The determination was carried out with a Phenomenex Kinetex C₁₈ column (100 mm×2.1 mm, 1.7 μm) as the stationary phase, gradient elution was carried out with methanol-0.1% formic acid aqueous solution as the mobile phase, ¹³C₃ acrylamide as the internal standard, and multiple reaction monitoring (MRM) mode was used to detect the residual acrylamide.

The content ranges of acrylamide in 247 batches of samples were 0-26 403 μg·kg^-1. These contents exceeded the regulations stipulated by some nations or organizations, and posed certain safety risks in long-term use.

The method established in this study is rapid, simple, sensitive and reproducible, and will lay a scientific basis for the quality control and safety evaluation of acrylamide in “8 famous medicines from Zhejiang” and other genuine medicinal materials.

The early warning of new psychoactive substance iso-propoxate (homologue of etomidate) was carried out through sewage drug monitoring technology, and quantitative research was conducted to provide technical support for management and law enforcement.

The abnormal results were obtained by quantitative analysis of multiple reaction monitoring (MRM) mode using online solid phase extraction-ultra high performance liquid chromatography tandem mass spectrometry (SPE-UPLC-MS/MS), and the basic information of abnormal substance was determined by precursor ion scanning mode. The qualitative determination was carried out using parallel reaction monitoring (PRM) mode by ultra high performance liquid chromatography quadrupole-orbitrap high resolution mass spectrometry (UPLC-Q exactive HRMS) after the directional synthesis of the compound. Quantitative study was then conducted by commercial reference products. The correlation analysis was carried out by SPSS 27.

The abnormal substance was determined to be iso-propoxate. The optimized MRM parameters could be distinguished from its isomers by specific ion abundance ratio. The correlation analysis of quantitative results showed that it was similar to etomidate and ketamine abuse areas.

In the process of sewage drug monitoring, it is necessary to pay attention to the proportional relationship between prototype and metabolite. It is important to summarize the characteristic fragments and characteristic neutral loss information of specific types of compounds, carry out screening non-targeted detection for key samples in key areas in the monitoring process, and analyze potential substances using high-resolution mass spectrometry and other methods, so as to fully leveraging the early warning role of sewage drug monitoring.

To establish an ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of daidzein, formononetin, genkwanin, quercetin, rutin, apigenin, kaempferol, luteolin and puerarin in Tongfu Jingyaotong tincture.

The Waters ACQUITY UPLC HSS T3 Column (100 mm×2.1 mm, 1.8 μm) was used with methanol and 0.1% formic acid (gradient elution) as the mobile phase at a flow rate of 0.3 mL·min^-1. Column temperature was 45 ℃, and injection volume was 2.0 μL. Electrospray ion source was adopted with positive and negative ion modes and multi-reaction monitoring and acquisition.

Apigenin, genkwanin and daidzein had good linearity in the range of 0.2-1 000 ng·mL^-1, puerarin, rutin, luteolin, kaempferol and formononetin in the range of 0.2-2 000 ng·mL^-1, and quercetin in the range of 1.0-5 000 ng·mL^-1. The resolution was good, and the RSDs of precision, repeatability, reproducibility and stability test were all below 5% (n=6). The average recoveries ranged from 82.7% to 111.8% with RSDs (n=6) of 2.0%-5.4%. The contents of daidzein, formononetin, genkwanin, quercetin, rutin, apigenin, kaempferol, luteolin and puerarin in 6 batches of Tongfu Jingyaotong tincture were 8.46-17.97 μg·mL^-1, 0.62-1.67 μg·mL^-1, 0.03-0.08 μg·mL^-1, 1.34-2.22 μg·mL^-1, 0.70-1.14 μg·mL^-1, 0.48-0.99 μg·mL^-1, 0.20-0.90 μg·mL^-1, 0.10-0.16 μg·mL^-1 and 167.95-227.75 μg·mL^-1, respectively.

This method has strong specificity, wide linear range, accuracy and efficiency, and can detect the contents of 9 active ingredients in Tongfu Jingyaotong tincture at the same time, which offers an effective quality control method for Tongfu Jingyaotong tincture.

To establish a simple and practical microbiological counting method suitable for sheet drug packaging materials by comparing the results of four sampling methods.

The samples of artificial contamination of sheet drug packaging materials were prepared by standard strain adding method. The surface microorganisms were collected by wiping method prescribed by national drug packaging container (material) standard, as well as by self-designed washing method, direct inoculation method and contact dish method. The corresponding recovery rate was calculated. The results of 4 sampling methods were analyzed by one way analysis of variance (ANOVA).

From high to low, the recovery rate was as follows: contact dish method, direct inoculation method, wiping method and washing method. There were significant differences in the recovery rates of contact dish method, wiping method and washing method (P<0.05). The recovery rate of contact dish method was slightly higher than that of direct inoculation method, but there was no significant difference between them (P>0.05). There was no significant difference in the recoveries of medicinal film, aluminum foil and hard sheet among the four methods (P>0.05).

The contact dish method is simple, sensitive and accurate, and can be used for the examination of the microbial limit counting method of sheet drug packaging materials.

To establish an HPLC characteristic chromatogram of Xifeng Huoluo capsules, and to determine the contents of gastrodin, amygdalin, strychnine, hydroxysafflower yellow A, brucine, paeoniflorin, ferulic acid, salvianolic acid B, cryptotanshinone, tanshinone Ⅰ, and tanshinone ⅡA.

Extraction of 75% methanol was analyzed by a Venusil MP C₁₈ chromatographic column (250 mm×4.6 mm, 5 μm) using 0.1% phosphoric acid (A) -acetonitrile (B) as the mobile phase with gradient elution at a flow rate of 1.0 mL·min^-1. The column temperature was 30 ℃. The detection wavelengths were variable. The characteristic chromatograms of 10 batches of Xifeng Huoluo capsules were evaluated by similarity evaluation, and the eleven identified index components were quantitatively determined.

There were thirty-one common peaks in the characteristic chromatograms of eleven batches of samples and the similarities were all above 0.99. The common peaks could ascribed to twelve medicinal materials, and eleven of them were identified. Eleven components showed good linearity within their respective ranges (r≥0.999 4), and the average recoveries were 99.3%-103.0% with RSDs of 1.5%-2.4%.

The established method has high sensitivity and strong specificity. The HPLC characteristic chromatogram combined with multi-component quantitative determination can fully reflect its inherent quality and can be used for the quality control of Xifeng Huoluo Capsules.

To optimize the content determination method of morphine sulfate suppositories, which had high toxicity of extraction solvent and poor specificity and generality of chromatographic conditions.

The optimization of the preparation method for the test solution was to prepare the test solution by ice bath freezing and n-heptane extraction, and to compare the content determination results of the test solution prepared by the United States Pharmacopeia method (extracted with trichloromethane). The optimization of chromatographic conditions for content determination was based on the comprehensive comparison of United States Pharmacopeia, British Pharmacopeia and YBH specification. Symmetry C₁₈ column (150 mm×4.6 mm, 5 μm) was finally used. The mobile phase was 0.01 mol·L^-1 potassium dihydrogen phosphate solution containing 0.202% sodium heptane sulfonate (containing 0.1% triethylamine, adjusted to pH 2.5 with phosphoric acid) -methanol (70∶30). The flow rate was 1.0 mL·min^-1. The column temperature was 30 ℃. The detection wavelength was 284 nm. The injection volume was 20 μL.

The content measured by ice bath freezing method was relatively low, while the content measured by n-heptane extraction and trichloromethane extraction was basically consistent, indicating that n-heptane could be used as extraction solvent instead of halogenated alkane. The linear range of morphine sulfate was 2.12-105.94 μg·mL^-1(r=0.999 9), and the LOQ was 10.60 ng. The RSD values of system precision, repeatability, intermediate precision and stability were all less than 2%. The average recoveries were 98.6%-99.6% and RSD was 0.21%-0.66%. After forced degradation, the main peak of the tested solution could be separated from the degraded impurities, with good specificity. By changing the flow rate, column temperature, pH value, mobile phase ratio and column brand, the main peak could be well separated from each impurity peak, with good robustness.

This study optimized the determination method for morphine sulfate suppositories. The preparation of the test solution used n-heptane instead of the highly toxic haloalkane, and meanwhile improved the chromatographic conditions, enhanced the environmental protection, specificity and applicability of the method. It can be used for the content determination of morphine sulfate suppositories.

To study the toxicity of heavy metal cadmium and screen detoxification drugs base on Caenorhabditis elegans.

The microfluidics chip technology was used as the drug screening platform. The 96-well plate exposure pretest was used, and K-medium blank group was set up. The exposure group consisted of nematodes exposed to heavy metal cadmium at the environmental relevant concentration of 0.25-15.0 μg·mL after homogenization. Vitamin C and calcium disodium ethylenediamine tetraacetate were selected as detoxification group. （10±2） nematodes were added into each orifice plate. The number of nematodes was observed and recorded under a microscope, and paid attention to the changes in the structure of the vulva. The experimental data were processed and analyzed by origin 2019b software.

Compared with blank group, the fatality rates of C. elegans in exposed group were respectively 0%, 1.67%, 4.76%, 67.46%, 100% and 100% at 0.25, 1.50, 5.0, 10.0, 12.5, 15.0 μg·mL^-1 concentration, compared with blank group. The structure of the nematode vulva was deformed, from slight protrusion to severe protrusion, and even a tuberous protrusion, and finally ruptured. The mortality of detoxification group was reduced to different degrees compared with exposure group.

Compared with blank control group, with the increase of cadmium exposure concentration and exposure time, cadmium can inhibit the survival of C. elegans, and damage the structure of C. elegans vulva to varying degrees. The results of detoxification group showed that vitamin C had little detoxification effect on cadmium. The complexation of calcium disodium ethylenediamine tetraacetate with cadmium could prolong the survival time of C. elegans to a large extent.

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.