Traditional Chinese Medicine Samples Research Articles

Recent advances in green solvents-based liquid-phase microextraction techniques for chromatographic analysis of active components in traditional Chinese medicine.

Traditional Chinese medicine (TCM) is a treasure of China and a crucial part of traditional medicine in the world, particularly in many oriental countries. TCM is the core and basis of traditional medicine in clinical practice for numerous diseases, and performs important function in nutraceuticals and dietary supplements. However, it is extremely difficult to extract each active ingredient from TCM to elucidate the mechanism of TCM clinical efficacy due to numerous compounds in TCM, especially trace compounds. Consequently, liquid-phase microextraction (LPME) techniques, one of the main extraction methods of active ingredients in TCM, have attracted a considerable attention from researchers. In recent years, many novel green solvents based-LPME methods have been reported, such as single-drop microextraction (SDME), hollow-fiber liquid-phase microextraction (HF-LPME), dispersive liquid-liquid microextraction (DLLME), and electro-membrane extraction (EME). Therefore, in this review, we present an up-to-date and comprehensive summary of various LPME techniques, novel green solvents, and their applications in the analysis of active ingredients within the complex TCM samples. We provide a detailed overview of the fundamental principles, modes, and the critical process parameters of the LPME techniques. In addition, we compare different types of green solvents (i.e., deep eutectic solvents, ionic liquids, magnetic ionic liquids, supramolecular solvents, switchable solvents, among others), and the advantages and disadvantages of these solvents are critically evaluated, highlighting their suitability for various applications. Finally, we elucidate the merits and demerits of different LPME methods, discuss their practical applications, and explore their future research directions. This review aims to provide a valuable resource for researchers and practitioners in the field of TCM, promoting research development and application of the advanced and environmentally friendly sample pretreatment techniques.

Application of Polydopamine-Based Magnetic Solid-Phase Extraction for Highly Sensitive Determination of Aristolochic Acid I from Traditional Chinese Medicine Samples.

A low cost-effective and simple synthesis method combining magnetic solid-phase extraction (MSPE) and high-pressure liquid chromatography was developed for the analysis of aristolochic acids I (AAI) in traditional Chinese medicine samples. A novel polydopamine (PDA) modified magnetic nanoparticles with one single carbon layer (Fe3O4@1C NPs) via one-pot hydrothermal approach was prepared and then successfully employed to extract AAI for the first time. Dopamine (DA) can form a PDA layer on Fe3O4@1C NPs surface through self-polymerization to form Fe3O4@1C@PDA. As a surface modifier of DA, PDA offered more adsorption sites to AAI due to π-π stacking, hydrogen bonding and electrostatic interactions. The parameters of MSPE were optimized by univariate and multivariate methods (Box-Behnken design) in detail. High degree of linearity was obtained in the range of 0.05-200.0μg/mL. The limits of detection (S/N = 3) and quantification (S/N = 10) were 0.08 and 0.25μg/mL, respectively. The recoveries of AAI in spiked Xiaoqinglong mixture samples were in the range of 86.7 to 108.5% with the relative standard deviation of less than 5.2%. Thus, a fast, convenient, sensitive and eco-friendly method was successfully proposed and became a promising approach for the determination of AAI in herbal plants or its preparation in the manufacturing procedure.

Cu,Ce-containing phosphotungstates as laccase-like nanozyme for colorimetric detection of Cr(VI) and Fe(Ⅲ)

Herein, a nanocomposite of Cu,Ce-containing phosphotungstates (Cu,Ce-PTs) with outstanding laccase-like activity was fabricated via a one-pot microwave-assisted hydrothermal method. Notably, it was discovered that both Fe3+ and Cr6+ could significantly enhance the electron transfer rates of Ce3+ and Ce4+, along with generous Cu2+ with high catalytic activity, thereby promoting the laccase-like activity of Cu,Ce-PTs. The proposed system can be used for the detection of Fe3+ and Cr6+ in a range of 0.667–333.33 μg/mL and 0.033–33.33 μg/mL with a low detection limit of 0.135 μg/mL and 0.0288 μg/mL, respectively. The proposed assay exhibits excellent reusability and selectivity and can be used in traditional Chinese medicine samples analysis.

Fabrication of amino- and hydroxyl dual-functionalized magnetic microporous organic network for extraction of zearalenone from traditional Chinese medicine prior to the HPLC determination

Efficient enrichment of trace zearalenone (ZEN) from the complex traditional Chinese medicine (TCM) samples is quite difficult, but of great significance for TCM quality control. Herein, we reported a novel magnetic solid phase extraction (MSPE) strategy for ZEN enrichment using the amino- and hydroxyl dual-functionalized magnetic microporous organic network (Fe3O4@MON-NH2-OH) as an advanced adsorbent combined with the high-performance liquid chromatography (HPLC) determination. Efficient extraction of ZEN was achieved via the possible hydrogen bonding, hydrophobic, and π-π interactions between Fe3O4@MON-NH2-OH and ZEN. The adsorption capacity of Fe3O4@MON-NH2-OH for ZEN was 215.0 mg g−1 at the room temperature, which was much higher than most of the reported adsorbents. Under the optimal condition, the developed Fe3O4@MON-NH2-OH-MSPE-HPLC method exhibited wide linear range (5–2500 μg L−1), low limits of detection (1.4–35 μg L−1), less adsorbent consumption (5 mg), and large enhancement factor (95) for ZEN. The proposed method was successfully applied to detect trace ZEN from 10 kinds of real TCM samples. Conclusively, this work demonstrates the Fe3O4@MON-NH2-OH can effectively extract trace ZEN from the complex TCM matrices, which may open up a new way for the application of MONs in the enrichment and extraction of trace contaminants or active constituents from the complex TCM samples.

A fluorescence method based on malachite green/aptamer for detection of ochratoxin A in traditional Chinese medicines

A label-free fluorescence method based on malachite green/aptamer was developed for the detection of ochratoxin A(OTA) in traditional Chinese medicines. Malachite green itself exhibits weak fluorescence. Upon interaction with the aptamer specific to OTA, the G-quadruplex structure of the aptamer provides a protective microenvironment for malachite green, which significantly enhances its fluorescence signal. After OTA is added, preferential binding occurs between the aptamer and OTA, and malachite green will be released from the aptamer, which weakens the fluorescence signal. According to this principle, this paper established a fluorescence method with the aptamer of OTA as the recognition element and malachite green as the fluorescent probe for the detection of OTA in traditional Chinese medicines. The key experimental factors such as the concentrations of metal ions, aptamer, and malachite green were optimized to improve the performance of the method. OTA was detected under the optimal experimental conditions, and the results showed that with the increase in OTA concentration, the fluorescence signal gradually weakened. Within the range of 20-1 000 nmol·L~(-1), the OTA concentration was linearly correlated with the fluorescence signal ratio ΔF/F(ΔF=F_0-F, where F_0 is the fluorescence signal of aptamer/malachite green, and F is the fluorescence signal of OTA/aptamer/malachite green), with R~2 of 0.995. The limit of detection of the established method was 7.1 nmol·L~(-1). Furthermore, three substances structurally similar to OTA and two mycotoxins that may coexist with OTA were selected for experiments, which aimed to examine the cross-reactivity and specificity of the established method. The cross-reactivity experiments demonstrated that the interferers did not significantly affect the fluorescence signal of the detection system. The specificity experiments revealed that when mycotoxins were mixed with OTA, the fluorescence signal generated by the mixture closely resembled that of OTA itself. The results indicated that even in the presence of interferents, the established method remained unaffected and demonstrated excellent specificity. Additionally, this method exhibited remarkable reproducibility and stability. In the case of simple centrifugation and dilution of traditional Chinese medicine samples(Puerariae Lobatae Radix, Sophorae Flavescentis Radix, and Periplocae Cortex), the OTA detection method was applicable, with recovery rates ranging from 91.5% to 121.3%. Notably, this approach does not need complex pretreatment of traditional Chinese medicines while offering simple operation, low detection costs, and short detection time. Furthermore, by incorporating aptamers into the quality evaluation of traditional Chinese medicines, this method expands the application scope of aptamers.

DNA barcoding combined with high-resolution melting analysis to discriminate rhubarb species and its traditional Chinese patent medicines.

Introduction: Rhubarb is a frequently used and beneficial traditional Chinese medicine. Wild resources of these plants are constantly being depleted, meaning that rhubarb products have been subjected to an unparalleled level of adulteration. Consequentially, reliable technology is urgently required to verify the authenticity of rhubarb raw materials and commercial botanical drugs. Methods: In this study, the barcode-DNA high-resolution melting (Bar-HRM) method was applied to characterize 63 rhubarb samples (five Polygonaceae species: Rheum tanguticum, Rh. palmatum, Rh. officinale, Rumex japonicus and Ru. sp.) and distinguish the rhubarb contents of 24 traditional Chinese patent medicine (TCPM) samples. Three markers, namely ITS2, rbcL and psbA-trnH, were tested to assess the candidate DNA barcodes for their effectiveness in distinguishing rhubarb from its adulterants. A segment from ITS2 was selected as the most suitable mini-barcode to identify the botanical drug rhubarb in TCPMs. Then, rhubarbs and TCPM samples were subjected to HRM analysis based on the ITS2 barcode. Results: Among the tested barcoding loci, ITS2 displayed abundant sites of variation and was effective in identifying Polygonaceae species and their botanical origins. HRM analysis based on the ITS2 mini-barcode region successfully distinguished the authenticity of five Polygonaceae species and eight batches of TCPMs. Of the 18 TCPM samples, 66.7 % (12 samples) were identified as containing Rh. tanguticum or Rh. officinale. However, 33.3 % were shown to consist of adulterants. Conclusions: These results demonstrated that DNA barcoding combined with HRM is a specific, suitable and powerful approach for identifying rhubarb species and TCPMs, which is crucial to guaranteeing the security of medicinal plants being traded internationally.

In-situ growth of metal coordination-synergistic imprinted polymer onto shrimp shell-derived magnetic FeNi biochar for specific recognition of monocrotaline in herbal medicine

In this work, an in-situ growth of metal coordination-synergistic imprinted polymer onto magnetic FeNi biochar was synthesized and further used for the recognition and isolation of toxic monocrotaline (Mct) from herbal medicines. The shrimp shell-derived iron nickel oxide doped magnetic biochar (INOMB) was successfully prepared by a combined impregnation and pyrolysis method. The molecularly imprinted polymers (MIPs) layer was then in-situ coated onto the surface of the INOMB carrier by the incipient metal coordination interaction between Mct and bimetallic Fe/Ni elements, and dopamine self-autoxidation process. The obtained FeNi@Mct-MIPs was systematically characterized through various characterization techniques. Profiting from the synergistic effect, the synthesized FeNi@Mct-MIPs obviously enhanced the adsorption capacity in capturing Mct, and the maximum adsorption capacity and imprinting factor were determined as 64.59 mg g−1 and 2.16 accordingly. Furthermore, the resultant FeNi@Mct-MIPs composite can be further used as a novel solid-phase extraction absorbent to enrich and detect trace Mct in herbal plants by using ultra-high performance liquid chromatography combined with mass spectrum (UHPLC-MS) technology. Overall, the present INOMB-based composite shows great potential for selective separation and detection of Mct from traditional Chinese medicine samples.

The United States dried seahorse trade: A comparison of traditional Chinese medicine and ecommerce-curio markets using molecular identification.

Tens of millions of dried seahorses (genus Hippocampus) are traded annually, and the pressure from this trade along with their life history traits (involved parental care and small migration distances and home ranges) has led to near global population declines. This and other forms of overexploitation have led to all seahorse species being listed in Appendix II under the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). The signatory nations of CITES recommended a 10-cm size limit of seahorses to ensure harvested individuals have reached reproductive maturity, and have thus had the chance to produce offspring, to maintain a more sustainable global seahorse fishery. We assessed adherence to CITES recommendations using DNA barcoding and size measurements to compare two prominent U.S. dried seahorse markets: (1) traditional Chinese medicine (TCM), and (2) non-medicinal ecommerce and coastal curio (ECC). We also estimated U.S. import abundance from CITES records. Of the nine species identified among all samples (n = 532), eight were found in the TCM trade (n = 168); composed mostly (75%) of the Indo-Pacific species Hippocampus trimaculatus, and Hippocampus spinosissimus, and the Latin American Hippocampus ingens. In contrast, ECC samples (n = 344) included 5 species, primarily juvenile Indo-Pacific Hippocampus kuda (51.5%) and the western Atlantic Hippocampus zosterae (40.7). The majority of TCM samples (85.7%) met the CITES size recommendation, in contrast to 4.8% of ECC samples. These results suggest non-size discriminatory bycatch is the most likely source of imported ECC specimens. In addition, CITES records indicate that approximately 602,275 dried specimens were imported into the U.S. from 2004-2020, but the exact species composition remains unknown as many U.S. imports records list one species or Hippocampus spp. from confiscated shipments due to difficulties in morphological identification and large numbers of individuals per shipment. Molecular identification was used to identify the species composition of confiscated shipment imports containing undesignated species, and similar to TCM, found H. trimaculatus and H. spinosissimus the most abundant. By combining DNA barcoding, size comparisons, and CITES database records, these results provide an important glimpse into the two primary dried U.S. seahorse end-markets, and may further inform the conservation status of several Hippocampus species.

Rational design of a ratiometric fluorescent aptasensor for patulin in traditional Chinese medicine through the studies of the interaction mechanism between its DNA aptamer and the target molecule.

Patulin (PAT) is a kind of mycotoxin which must be monitored for the sake of quality and safety in traditional Chinese medicine (TCM) owing to its harm to human health. On this account, a rationally designed ratiometric fluorescent aptasensor was developed based on the studies of the interaction mechanism between PAT and its aptamer (PAT-APT). First, CD spectroscopy, molecular docking, and molecular dynamic simulation were applied to investigate the details on how PAT-APT binds with its target molecule. The results indicated that the structure of PAT-APT changed to a certain extent and was stabilized after binding with PAT. C-11, C-37 and C-38 were the key sites for the recognition and interaction between PAT-APT and its target. Second, based on these results, a ratiometric aptasensor was designed using fluorescence resonance energy transfer (FRET) and synchronous fluorescence spectroscopy. A complementary sequence (cDNA) to the aptamer with an appropriate length and hybridization position was obtained through rational design and optimization. Both PAT-APT and cDNA were labeled using a pair of fluorophores, which could generate FRET when the two single-stranded oligonucleotides hybridized. The accurate detection of PAT could be realized according to the change ratio of the fluorescence intensity at the corresponding wavelengths of the two fluorophores before and after the assay. The aptasensor achieved an ultralow limit of detection of 0.16 nM, perfect selectivity, and satisfactory practicability in complex TCM samples. To our knowledge, this is the first aptasensor for PAT designed through the interaction mechanism between its aptamer and the target molecule. Moreover, the assay for PAT is cost-effective, does not need complicated pretreatment and only takes less than an hour. In summary, this study makes a contribution to the safety control of TCM and provides a thinking mode from mechanism to rational design to conquer the problem of sensitive aptasensing of one component in a complex system.

Colorimetric assay based on peroxidase-like activity of dodecyl trimethylammonium bromide-tetramethyl zinc (4-pyridinyl) porphyrin for detection of organophosphorus pesticides.

A simple and sensitive colorimetric assay for detecting organophosphorus pesticides (OPs) was developed based on 3,3',5,5'-tetramethylbenzidine (TMB)/hydrogen peroxide (H2O2)/dodecyl trimethylammonium bromide (DTAB)-tetramethyl zinc (4-pyridinyl) porphyrin (ZnTPyP). In this system, based on the peroxidase-like activity of DTAB-ZnTPyP, H2O2 decomposes to produce hydroxyl radicals, which oxidize TMB, resulting in blue oxidation products. The OPs (trichlorfon, dichlorvos, and thimet) were first combined with DTAB-ZnTPyP through electrostatic interactions. The OPs caused a decrease in the peroxidase-like activity of DTAB-ZnTPyP due to spatial site blocking. At the same time, π-interactions occurred between them, and these interactions also inhibited the oxidation of TMB (652 nm), thus making the detection of OPs possible. The limits of detection for trichlorfon, dichlorvos, and thimet were 0.25, 1.02, and 0.66 μg/L, respectively, and the corresponding linear ranges were 1-35, 5-45, and 1-40 μg/L, respectively. Moreover, the assay was successfully used to determine OPs in cabbage, apple, soil, and traditional Chinese medicine samples (the recovery ratios were 91.8-109.8%), showing a great promising potential for detecting OPs also in other complex samples.

A piecewise mirror extension local mean decomposition method for denoising of near-infrared spectra with uneven noise

As a new signal analysis tool, local mean decomposition (LMD) can decompose a spectrum into a set of single-component signals with different frequencies. However, LMD may produce endpoint effect. At the same time, due to instrumental instability and experimental errors, uneven noise may occur in the measurement process of near-infrared (NIR) spectra. A piecewise mirror extension local mean decomposition (PME-LMD) method is proposed for denoising of NIR spectrum with uneven noise. Firstly, the NIR spectrum with uneven noise is segmented to obtain several intervals. Then, each interval is mirrored extension to the left and right for overcoming the endpoint effect. Subsequently, a series of product functions (PFs) are obtained by LMD for each piecewise mirror extended NIR spectrum. Finally, the denoised NIR spectrum is obtained by intercepting and reconstructing the components without noise. One artificial noised signal and NIR spectra of traditional Chinese medicine (TCM) and beer samples are used to validate the performance of this method. As comparison to PME-LMD method, Savitzky-Golay (SG) smoothing, discrete wavelet transform (DWT), empirical mode decomposition (EMD) and LMD have also been investigated. Visualization spectra, signal-to-noise ratio (SNR) and modeling results of partial least squares (PLS) are used as evaluation indicators. The results demonstrate that the proposed method performed best in terms of SNR, root mean squared error of prediction (RMSEP) and correlation coefficient (R) of PLS. Furthermore, the piecewise step is critical to perform LMD denoising for NIR spectrum with uneven noise. At the same time, the endpoint effect of LMD is effectively overcome by mirror extension.

Characterization of effective phytochemicals in traditional Chinese medicine by mass spectrometry.

Traditional Chinese medicines (TCMs) have been widely used in clinical and healthcare applications around the world. The characterization of the phytochemical components in TCMs is very important for studying the therapeutic mechanism of TCMs. In the analysis process, sample preparation and instrument analysis are key steps to improve analysis performance and accuracy. In recent years, chromatography combined with mass spectrometry (MS) has been widely used for the separation and detection of trace components in complex TCM samples. This article reviews various sample preparation techniques and chromatography-MS techniques, including the application of gas chromatography-MS and liquid chromatography-MS and other MS techniques in the characterization of phytochemicals in TCM materials and Chinese medicine products. This article also describes a new ambient ionization MS method for rapid and high-throughput analysis of TCM components.

Elucidation of aroma compounds in passion fruit (Passiflora alata Ait) using a molecular sensory approach.

In this experiment, Guangxi passion fruit was used as the raw material for natural aroma extraction using the spinning cone column (Spinning Cone Column, SCC) technique. In combination with the semi-quantitative method, the aroma characteristics of the raw pulp (raw whole-fruit puree, PU) before SCC processing, residue (Residue, RS) and extract (Extract, EX) after SCC processing, and passion fruit juice (Juice, JU) were evaluated for their aroma characteristics using headspace gas chromatography-mass spectrometry (HS-SPME-GC-MS), gas chromatography-ion mobility spectrometry (GC-IMS), electronic nose, and sensory evaluation. As a result, a total of 110 aroma substances were detected in four samples, and 33, 38, 73, and 28 aroma components were detected from PU, RS, EX, and JU, respectively. There are 50 compounds in EX with concentrations greater than 10 μg/kg, and 19 of them had OAV values greater than 1, including β-Ionone and linalool, which contributed significantly to the aroma. The aroma profiles and characteristics were further analyzed for JU and EX using the e-nose sensor, and it was found that both showed similar aroma profiles. The sensory evaluation results were also in general agreement with the results obtained from the electronic nose, with EX having mainly "floral", "fruity," and "sweet" aromas. The results demonstrated that the spinning cone column technique can increase the fresh and natural fruity aroma of passion fruit in the extract, which has the effect of enriching the aroma and improving the aftertaste. This study will make a foundation for passion fruit SCC extract application in drinks. PRACTICAL APPLICATIONS: Compared with traditional extraction technology, spinning cone column technology has the advantages of high mass transfer efficiency, short extraction time, a wide range of temperature control, and the most complete extracted flavor substances, which greatly reduces the damage degree of heat-sensitive flavor substances and condense aroma. It is widely used in beverages, wine, dairy products, fruit and vegetable, spice essential oil, and other industries. Passion fruit flavor prepared by SCC technology has the advantages of high purity and high concentration, which can be used in solid drinks, baked food, convenience food, tobacco, perfume, and other products. Besides, GC-IMS is an efficient and rapid new analytical technique, which has been widely used in the flavor analysis of volatile organic compounds in food and traditional Chinese medicine samples.

Preparation of molecularly imprinted foam for selective extraction of toxic monocrotaline from herbs

Monocrotaline (MCT), derived from most flowering plants, showed significant hepatotoxicity and carcinogenicity. Humans are easily exposed to MCT by eating traditional herbs or contaminated foods, posing a huge threat to human health. In order to selectively and conveniently separate and enrich MCT from these complex samples, we fabricated three-dimensional porous and homogeneous molecularly imprinted polymer foams (MIPFs) by using considerably inexpensive polyurethane foam (PU) as the carrier. The morphology, stability, adsorption properties and extraction parameters of MIPFs were investigated. The results indicated that an imprinted layer was coated on the surface of the carrier; the stability of MIPFs was excellent; In addition to hydrogen bonding and spatial complementarity, the electrostatic interactions were crucial for the recognition between MCT and MIPFs; MIPFs exhibited high adsorption capacity of 22.78 mg g−1, fast mass transfer rate, and satisfactory selectivity for MCT. Subsequently, MIPFs were applied as the solid phase extraction (SPE) absorbents and combined with high performance liquid chromatography (HPLC) to enrich and detect MCT in herbal plants. The results showed that MCT could be efficiently enriched, and the impurities could be dramatically reduced. MIPFs hold great potential for selective separation and detection of MCT in complex matrices, such as traditional Chinese medicine samples and food samples.

Rapid and interference-free quantification of nine coumarins in Cnidii fructus using HPLC-DAD assisted with second-order calibration model

High performance liquid chromatography-diode array detection (HPLC-DAD) assisted with second-order calibration model based on alternating trilinear decomposition-assisted multivariate curve resolution (ATLD-MCR) model was proposed to quantify nine coumarins in traditional Chinese medicine (TCM) samples of Cnidii Fructus. This advanced analytical strategy shows good linearity (r > 0.9991), rapidity (t < 10.50 min) and accuracy, in low chromatographic resolution data even in the presence of potential interfering species. Satisfactory qualitative and quantitative results were achieved in the presence of complex matrix interferences from the TCM samples and the second-order advantage was fully exploited. The average spiked recoveries for most target analytes are in the range of 84.1%–108.0% with the standard deviations less than 6.9%. Furthermore, the quantitative results were further compared with a classical univariate calibration applied to a series of peak area data acquired from HPLC-UV detection. Statistical parameters and analytical figures of merit were calculated according to modern analytical protocols to validate the accuracy of the proposed analytical method. The results show that the proposed analytical method had the advantages of high-efficiency, low-cost and met the requirements of green analytical chemistry. Therefore, it promises to be a potential and universal way for qualitative and quantitative analysis of coumarins in pharmaceutical and food fields.

Simultaneous determination of 76 pesticide residues in the traditional Chinese medicine by magnetic hydrophilic-lipophilic-balanced materials assisted matrix solid phase dispersion extraction-high performance liquid chromatography-tandem mass spectrometry

建立灵敏、可靠的中药材中农药多残留的检测方法对保证中药材的质量和安全十分重要。制备了磁性亲水亲脂平衡萃取材料Fe3O4@PLS,将其应用于农药多残留的磁性基质固相分散萃取中,并结合高效液相色谱-串联质谱法(HPLC-MS/MS)检测了金银花、菊花和三七块根(干)3种中药材中76种农药残留量。研究通过扫描电子显微镜(SEM)、傅里叶变换红外光谱(FT-IR)和X-射线衍射仪(XRD)对磁性萃取材料Fe3O4@PLS进行表面形貌和结构的表征。同时考察了影响磁性基质固相分散萃取效率的主要因素,结果表明,磁性萃取材料Fe3O4@PLS的用量为10 mg、研磨分散吸附时间为5 min、淋洗液为10 mL 20%(v/v)甲醇水溶液、涡旋振荡清洗1 min、以0.5 mL 0.1%(v/v)甲酸乙腈为洗脱剂、涡旋振荡洗脱1 min, 76种农药的萃取效果最佳。在实际应用中,76种农药在金银花、菊花、三七块根(干)3种中药材中的萃取回收率别为69.1%~112.2%、67.1%~102.8%和70.1%~105.1%,相对标准偏差分别为2.0%~12.4%、2.1%~13.2%和2.0%~13.5%。该方法利用Fe3O4@PLS良好的磁响应性和亲水亲脂通用吸附特性,可以同时萃取极性农药(如多菌灵等)和非极性农药(如敌瘟磷等),建立了测定中药材中76种农药残留的磁性基质固相分散萃取-高效液相色谱-串联质谱联用的分析方法,具有低消耗、操作简便、灵敏度高等优点,适用于非液态中药材基质中多种类农药残留的检测。

Fingerprint of Shenmai Injection based on ~1H-NMR technique

Shenmai Injection is a Chinese medicinal injection prepared from Ginseng Radix et Rhizoma Rubra and Ophiopogonis Radix, which is widely used in clinical practice for the treatment and adjuvant therapy of cardiovascular diseases with significant pharmacological effects. Proton nuclear magnetic resonance spectroscopy(~1H-NMR) has the advantages of simple and nondestructive sample pretreatment, fast analysis, abundant chemical information, quantification and no need to follow the standard curve. It is widely used in the analysis and research of complex mixtures of traditional Chinese medicine, clinical blood and urine samples. In this study, the ~1H-NMR fingerprint of Shenmai Injection was established. Thirty-two chemical components were identified, including seven amino acids, eight small molecular organic acids, one alkaloid, four sugars, two nucleosides, seven saponins, and three other components. Pearson's correlation coefficient and multivariate analysis of variance(principal component analysis combined with hierarchical cluster analysis) were applied based on the ~1H-NMR fingerprint to evaluate the quality consistency. The results showed high-quality consistency of 82 batches of Shenmai Injection. This study confirms that the ~1H-NMR fingerprint has great potential in the application of quality control of Chinese medicinal injection.

Conductive polymer sorbent for extraction and determination of resveratrol and polydatin in Polygonum cuspidatum root samples.

The quantitative analysis of trace resveratrol and polydatin in plant tissues is suitable for elucidation of the compounds' mechanisms of action. The main objective of this work was to develop a feasible and effective sample pretreatment method to measure the concentrations of resveratrol and polydatin in complex samples. A polymer sorbent, poly(2-mercaptobenzimidazole), was electrochemically prepared and utilized for selective extraction, while resveratrol and polydatin were used as target analytes. The sorbent was characterized by cyclic voltammetry, scanning electron microscopy and Fourier transform infrared spectroscopy. After extraction and elution, the analytes were analyzed by a Thermo U3000 HPLC system. Several affecting parameters, including the volume of elution solution, sample pH value, sample flow rate and sample volume, were evaluated and optimized. The proposed method showed good linearity with low limits of detection (from 0.5 to 0.8ng·mL-1 ) and ideal accuracy with spiked recoveries from 81.30% to 99.16%. A good enrichment factor (more than 200-fold) together with good sensitivity was obtained with this method. Analysis of resveratrol and polydatin in Polygonum cuspidatum samples by this method is efficient. The method developed in this work exhibits several significant merits, including easy operation and high extraction efficiency, indicating that electrochemically prepared polymer sorbent is useful for sample pretreatment and analysis of traditional Chinese medicine samples.

A fluorescent aptasensor based on gold nanoparticles quenching the fluorescence of rhodamine B to detect acetamiprid.

Pesticide residue detection is one of the main safety issues in the utilization of medicinal plants. In this work, a highly selective and sensitive aptasensor for acetamiprid determination was designed. The mechanism of the proposed method is based on the fluorescence resonance energy transfer (FRET) between gold nanoparticles (AuNPs) and rhodamine B (RB). Aptamers protect AuNPs from salt-induced aggregation, which causes fluorescence quenching of RB by the AuNPs via surface energy transfer. In the absence of acetamiprid, AuNPs were coated with aptamers on the surface and dispersed in NaCl solution. At this time, the dispersed AuNPs could perfectly quench the fluorescence intensity of RB. In contrast, in the presence of acetamiprid, aptamers specifically combine with acetamiprid to form a complex. With a high salt concentration, AuNPs would be aggregated without aptamer protection, weakening the RB quenching effect. Therefore, the concentration of acetamiprid could be obtained from the change in fluorescence intensity in the system. A fluorescent sensing method was established with a linear range from 0.1 to 3 μg mL-1, and the LOD was 0.0285 μg mL-1. The recoveries of acetamiprid in traditional Chinese medicine (TCM) samples were 96.23-105.75%. This method has great application value for the detection of acetamiprid in a complex sample matrix.

Development of an immunochromatographic test strip for the rapid detection of aristolochic acid A in herbal medicinal materials.

Plants containing aristolochic acid and its derivatives are nephrotoxic, mutagenic, and carcinogenic to humans; chronic diet poisoning caused by the aristolochic acid is the cause of endemic (Balkan) nephropathy and related cancers. To develop a colloidal gold immunochromatographic test strip (ICS) based on the competitive format for the rapid detection of aristolochic acid A (AA-A) in herbal medicinal materials. For the ICS based on gold nanoparticles (AuNPs), the antigen [AA-A-bovine serum albumin (BSA)], and goat anti-mouse IgG were drawn on the nitrocellulose membrane as the test line (T line) and the control line (C line), respectively. Monoclonal antibody (MAb)-AuNP conjugates were sprayed onto the conjugate pad. The sensitivity of the ICS was 6 ng/mL, and the test was completed in 10 min. The analysis of AA-A in traditional Chinese medicine samples showed that the ICS results were in good agreement with those obtained by high-performance liquid chromatography methods. These results demonstrated that the ICS test could be used as a reliable, rapid, cost-effective, and convenient qualitative tool for on-site screening techniques to detect AA-A in herbal medicinal materials without any special instrumentation.