Development and validation of a deep eutectic solvent-assisted liquid-liquid extraction method for simultaneous quantification of six steroid hormones in serum by liquid chromatography-tandem mass spectrometry.

Steroid hormones have been reported to be associated with endocrine system diseases. This paper proposes a novel procedure of deep eutectic solvent (DES)-assisted liquid-liquid extraction (LLE) to extract six steroid hormones (including cortisone, cortisol, androstenedione, testosterone, 17-hydroxyprogesterone, and progesterone) from serum coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of five types of L-proline, choline chloride, and citric acid-based DESs were tailored; the DES from L-proline and ethylene glycol at a molar ratio of 1:4 with 20 % acetonitrile was selected as the best-fit assisted solvent for the six steroid hormones compared with other DESs. The parameters for extraction by selected DES were optimized using Box-Behnken design (BBD), and the optimal extraction conditions are 200 µL of acetonitrile, 100 µL of the sample, and 80 µL of DES. Under optimum conditions, the method has good linear calibration ranges (between 0.07 ng mL-1 and 600 ng mL-1), correlation coefficients of determination (r2>0.99), and low limits of quantification (between 0.02 and 0.60 ng mL-1). The extraction recoveries were in the range of 81.84-114.43 %, and the intra-day and inter-day relative standard deviations (RSDs) were less than 10 %.In general, the DES-LC-MS/MS method is a simple and environmentally-friendly method, which can be complementary to the presently available methods for determining steroid hormones in serum.

Method validation for detection of afidopyropen and M440I007 in tea.

BACKGROUND: Afidopyropen is a novel biorational insecticide for controlling piercing pests with great potential for application in tea gardens that can form the metabolite M440I007 when utilized for crops. However, because of a lack of analytical method for afidopyropen and M440I007 in tea, there is no means of monitoring the residues. Therefore, method development, validation and simultaneous determination of afidopyropen and M440I007 in fresh tea leaves, dried tea and tea infusion is of prime significance. RESULTS: A TPT cartridge-based method was developed for the solid phase extraction of afidopyropen and M440I007 from tea matrices. Extraction and clean-up conditions, including the composition, volume and temperature of elutions, were optimized to achieve the best results. Both targets were extracted using water and acetonitrile, with a water:acetonitrile (v/v) ratio of 4:10 for fresh leaves and 8:10 for dried tea, which were then cleaned and analyzed using ultraperformance liquid chromatography-tandem mass spectrometry. Both analytes demonstrated excellent linearity with a correlation coefficient above 0.998. The optimized analytical method offered limits of quantifications of 0.005, 0.005 and 0.002 mg kg-1 (converted to dried tea) in fresh tea shoots, dried tea and tea infusion for both targets, respectively. Average recoveries of afidopyropen and M440I007 ranged from 79.0% to 101.5%, with relative standard deviations ≤ 14.7%. CONCLUSION: The results showed that the method of determination for these insecticides in tea matrices was practical and efficient. © 2023 Society of Chemical Industry.

Green and Efficient Determination of Fluoroquinolone Residues in Edible Green Fruits and Leafy Vegetables by Ultrasound-Assisted Extraction Followed by HPLC-MS/MS.

In this work, a simple, quick and efficient analytical method for determination of human and veterinary fluoroquinolone antimicrobial residues in lettuce, cucumber and spinach is developed. The procedure entails a 6 min ultrasound-assisted extraction (UAE, 3 × 2 min) in an alkaline (2% v/v NH3) aqueous solution containing Mg2+ ions (3 × 6 mL), with no need for organic solvents. The extract is submitted to cleanup on the HLB™ cartridge and the fluoroquinolones are separated and quantified by HPLC-MS/MS in a 10 min chromatographic run, using a small amount of acetonitrile in the mobile phase. The method, entirely developed in real matrices, is validated according to the updated analytical guidelines and provided suitable recoveries in the range of 67-116% and precision (RSD ≤ 20%, n = 3) at different concentrations (15, 70 and 150 ng g-1), with method quantification limits of 2-10 ng g-1. Fluoroquinolones were detected and quantified at concentrations from few to hundreds of nanograms per gram in vegetables from supermarkets, demonstrating the applicability of the method for monitoring residues of these pharmaceuticals.

Simple and rapid method for 336 multiresidual pesticide analysis in saliva, determination of their chemical stabilities, and biomonitoring of farmers.

Simultaneous multiresidual pesticide analysis of saliva samples was performed using scaled-down QuEChERS extraction with LC-MS/MS and GC-MS/MS. The optimum extraction procedure using acidified acetonitrile was applicable to 336 pesticides (287 for LC-MS/MS and 49 for GC-MS/MS). To determine pesticide multiresidues in saliva, 100 µL of the sample was extracted with 200 µL of 0.1% formic acid in acetonitrile, and the initial extract was partitioned with 40 mg of MgSO4 and 10 mg of NaCl. The organic supernatants (120 µL) were then mixed with acetonitrile (30 µL) for matrix-matching (4:1, v/v), and the final extract solution was injected into the LC-MS/MS (4 µL) and GC-MS/MS (2 µL) systems. The established analytical method showed a good LOQs between 5 and 25 ng/mL with reliable accuracy/precision values and recovery results (50-140%) for the target pesticides. Under the two different storage conditions, most of the analytes did not undergo chemical changes in the saliva samples, whereas some pesticides were more stable in freeze-thaw processes than those left at room temperature. Biomonitoring of farmers (ten mixers and ten sprayers) was successfully applied using the validated method, and two carbamates (fenobucarb and propamocarb) were determined at trace concentrations (12.5-675.0 ng/mL from 11 positively detected samples).

Acyl-Nω-methylserotonins and Branched-chain Acylserotonins in Lemon and Other Citrus Seeds-New Lipids with Antioxidant Properties and Potential Pharmacological Applications.

We have found 15 previously unknown compounds in seeds of lemon and other citrus species, such as tangerine, grapefruit and pomelo. The structure of these compounds was characterized by HR-MS spectrometry, fluorescence spectroscopy and chemical synthesis. These compounds were predominantly long-chain (C20-C25), saturated acyl-Nω-methylserotonins with the main contribution of C22 and C24 homologues, usually accounting for about 40% and 30% of all acylserotonins, respectively. The other, previously undescribed, minor compounds were branched-chain acylserotonins, as well as normal-chain acylserotonins, recently found in baobab seed oil. Within the seed, acylserotonins were found nearly exclusively in the inner seed coat, where probably their biosynthesis proceeds. On the other hand, lemon seedlings contained only trace amounts of these compounds that were not found in adult leaves. The compounds identified in the present studies were shown to have antioxidant properties in vitro, using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. In the investigated reaction in hexane, Me-C22 and Me-C24-serotonins were less active than n-C22 and n-C24-serotonins and δ-tocopherol, while branched-chain acylserotonins (iso-C21 and -C25) showed higher antioxidant activity than all the normal-chain compounds. On the other hand, all these compounds showed a similar but considerably lower antioxidant activity in acetonitrile than in hexane.

[Rapid determination of 22 functional components in cosmetics using ultra-high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry].

Plant components from extracts of Sophora flavescens, rhodiola, ginseng, Centella asiatica, and tea play important roles in skin whitening, moisturizing, anti-aging, sun protection, anti-inflammation, antiseptic, bacteriostatic, and other effects of cosmetics. At present, no relevant standard methods have been established to detect the addition amounts of plant extracts in cosmetics. In addition, plant extracts listed in product labels may be undetectable due to their addition in trace quantities and the lack of technical support. Therefore, a quantitative method for the simultaneous determination of 22 functional components in cosmetics was established by ultra-high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry (UHPLC-LTQ/Orbitrap MS). Target compounds were extracted with methanol from samples using ultrasonic extraction, and then separated on a C18 column (100 mm × 2.1 mm, 1.8 µm) with gradient elution of 0.1% (v/v) formic acid aqueous solution (A) and acetonitrile (B). The gradient elution program were as follows: 0-5 min, 5%B-8%B; 5-25 min, 8%B-60%B; 25-35 min, 60%B-80%B; 35-36 min, 80%B-5%B; 36-45 min, 5%B. The flow rate was 0.3 mL/min and the injection volume was 5 µL. Accurate masses of precursor ions were used to detect cosmetic functional components in positive ionization mode. The fragment ions obtained by higher energy collisional dissociation were used for confirmation of the functional components. Each compound showed good linearity. The limits of detection (LODs) were in the range of 0.003-2.01 mg/kg, and the limits of quantification (LOQs) were in the range of 0.02-4.36 mg/kg. Recoveries at three levels were 63.2%-125.1%, and relative standard deviations (RSDs) were 0.18%-10.9%. Fifty-four batches of samples labeled with four monomer functional components and nine plant extracts were tested. In the 17 batches of samples labeled with nicotinamide, 4 batches labeled with caffeine, and 6 batches labeled with Sophora flavescens root extract, the labeled functional components were detected. One out of 11 batches of samples labeled with D-panthenol was not detected. Three of the seven batches of samples labeled with ascorbyl glucoside were not detected. In the 21 batches of samples labeled with licorice extracts, the corresponding functional components were not detected in 9 batches. In the 21 batches of samples labeled with Centella asiatica extract, the corresponding functional components were not detected in 11 batches. In the 13 batches of samples labeled with tea extract, the corresponding functional components were not detected in 8 batches. In 11 of the 12 batches containing ginseng root extract, the corresponding functional components were not detected. In five of the six batches of astragalus membranaceus root extract samples, the corresponding functional components were not detected. In samples labeled with Polygonum cuspidatum root extract, Rehmannia glutinosa root extract, and Ophiopogon japonicus root extract, the corresponding functional components were detected. The method is simple, rapid, reliable, accurate, and suitable for the determination of the 22 functional components in cosmetics.

[Rapid screening of 84 pesticide residues in dendrobium by Sin-QuEChERS Nano purification column with gas chromatography-tandem mass spectrometry].

A rapid screening method for 84 pesticide residues in dendrobium perfringens parent material with different polarities was developed using a Sin-QuEChERS Nano clean-up column combined with gas chromatography-tandem mass spectrometry (GC-MS/MS). The differences in extraction efficiency of the targets were compared with different extraction solvents (acetonitrile containing 1% acetic acid, acetone) and methods (immersion with or without water). The purification effect and extraction recoveries of Sin-QuEChERS Nano method and classical dispersive solid-phase extraction (dSPE), solid-phase extraction (SPE) and QuEChERS were systematically compared using Dendrobium nobile samples. The differences in matrix effects between the Sin-QuEChERS Nano method, which was more effective in purification, and the dSPE method were also analyzed. The purification effects of three commercially available Sin-QuEChERS Nano purification columns (simple matrix purification column, complex matrix purification column and herbal purification column) were compared. The applicability of the purification methods were also verified by using different parts of Dendrobium nobile samples (stems, leaves and flowers). From the results, it could be concluded that weighing 2.00 g and the samples in 5 mL of water for 20 min, followed by extraction with acetonitrile containing 1% acetic acid was more effective. The average extraction recovery of the target components by Sin-QuEChERS Nano purification method was 90.5%, which further identified Sin-QuEChERS Nano-Chinese medicine purification column as the preferred purification column for dendrobium purification. The target components were separated by a DB-1701MS quartz capillary column (30 m×0.25 mm×0.25 µm) with programmed temperature rise, detected by multiple reaction monitoring (MRM) mode, and quantified by matrix-matched solution external standard method. The GC-MS/MS assay was used for the methodological validation of the 84 representative pesticides within Dendrobium officinale and Dendrobium nobile was carried out by GC-MS/MS detection method. The results indicated that the targets showed excellent linear correlation in different scopes with correlation coefficients (r2) >0. 990. The limits of detection (LODs, S/N=3) of the method were 1.5 to 5.8 µg/kg, and the limits of quantification (LOQs, S/N=10) ranged from 5.0 to 15.0 µg/kg. The spiked recoveries of the target pesticides under different spiked levels were 68.7%-116.2%, and the relative standard deviations (RSDs, n=6) were less than 15%. Compared to other typical pretreatment methods, the Sin-QuEChERS Nano method provided better performance in terms of purification. The method not only effectively removed pigments, organic acids, and alkaline interferents, but also saved preparation time. Losses due to solvent transfer were also avoided and no further vortexing or centrifugation was required, making it a simplified and effective extraction and purification procedure. The method was sensitive, rapid, simple and reliable. It effectively improved the detection efficiency during the rapid screening of pesticides in dendrobium and presented a strong practical application value. In addition, the developed method could further expand the types of target pesticides and could be used to detect more pesticide residues in foods and Chinese herbal medicine. The established Sin-QuEChERS Nano method was used for the analysis of authentic samples. The applicability of the method was evaluated by analyzing a total of 80 samples collected from Anlong, Libo, Dushan, and Yanhe County in Guizhou Province. The types of samples included dendrobium maple, Dendrobium nobile (flowers, stems, leaves) and Dendrobium officinale (flowers, stems, leaves, powder, tablets). At least one pesticide residue was detected in 12 samples, with a detection rate of 15%. The five pesticides with higher detection rates and residues were chlorpyrifos (0.08-0.5 mg/kg), chlorothalonil (0.06-3.2 mg/kg), propanil zinc (0.03-0.15 mg/kg), methyl parathion (0.04-0.23 mg/kg) and cyhalothrin (0.10-2.68 mg/kg). Except for the pesticides in maximum residue limits (MRLs), the pesticide residues detected from dendrobium samples were below the limits set by Chinese national standard (GB 2763-2021) and local standard DBS 52/048-2020.

Optimization of microwave assisted extraction of simmondsins and polyphenols from Jojoba (Simmondsia chinensis) seed cake using Box-Behnken statistical design.

A closed-vessel microwave-assisted extraction (MAE) of simmondsins and polyphenols from defatted Jojoba cake using Box-Benkhen design with four independent variables (solvent/cake ratio, ethanol concentration, extraction time and microwave power) was investigated. ANOVA results showed that the obtained models were significant at 95% confidence level. Optimal extraction conditions were found for highest values of microwave power (500 W) and extraction time (15 min) and for moderate values of solvent to cake ratio (41 - 45 mL/g). Optimum simmondsins yield (23.35%) was obtained with pure water as solvent. However, optimum polyphenols yield (2.33%) and ORAC antioxidant activity (656 µmol TE/g) were obtained with 46.79% and 42.04% ethanol in water, respectively. ORAC antioxidant activity was found to be well correlated to polyphenol and simmondsin contents. These results indicate that MAE is an effective technique for recovery of bioactive compounds for food and pharmaceutical industries from Jojoba by-products.

Effect of solvent and extraction technique on composition and biological activity of Lepidium sativum extracts.

The aim of this study was to evaluate chemical and biological potential of garden cress (Lepidium sativum L.) to receive valuable plant extracts with potential application in pharmacy or food industry. Four techniques of extraction and three environmentally friendly solvents such as water, supercritical CO2 and ethanol have been tested. Biological activity and chemical profile were evaluated in obtained extracts. GC/MS analysis showed that SFE extract from dried sprouts of L. sativum was especially rich in such glucosinolate derivatives as benzyl cyanide and benzyl thiocyanate. However, the extract obtained from freeze-dried sprouts by SFE with addition of 96% ethanol as co-solvent was especially rich in flavonoids and simultaneously exhibited the best antimicrobial activity. Comparison of MALDI-TOF-MS spectra of all obtained extracts clearly indicates that both SFE and maceration with water are the most selective techniques of extraction due to the lowest level of interfering substances with high molecular masses.

Simultaneous determination and risk assessment of metalaxyl and azoxystrobin in potato by liquid chromatography with tandem mass spectrometry.

A liquid chromatography with tandem mass spectrometry method was developed and validated to simultaneously determine metalaxyl and azoxystrobin in soil, potato, and potato foliage samples. The samples were extracted by 20 mL of acetonitrile and purified with dispersive solid-phase extraction using octadecyl silane as sorbent. The method showed good linearity (determination coefficients ≥ 0.9926) for metalaxyl (2.5-500 ng/mL) and azoxystrobin (5-1000 ng/mL). The limits of detection and quantification for both fungicides were 1.5-20 µg/kg. The average recoveries in soil, potato, and potato foliage were 83.07-92.87% for metalaxyl and 82.71-98.53% for azoxystrobin. The intra- and inter-day relative standard deviations were all less than 9%. The method was successfully applied on the residual analysis of metalaxyl and azoxystrobin in field trial samples. The results showed that the concentrations of metalaxyl and azoxystrobin in potato samples collected from Guizhou and Hunan were below 50 and 100 µg/kg (maximum residue limit set by China), respectively, at 5 days after the last application. When following the recommended application manual, metalaxyl and azoxystrobin do not present health concerns to the population because the risk quotients are far below 100%. All the above data could help and promote the safe and proper use of metalaxyl and azoxystrobin in potato.

Nitrogen compounds in Phacelia tanacetifolia Benth. honey: First time report on occurrence of (-)-5-epi-lithospermoside, uridine, adenine and xanthine in honey.

Lacy phacelia (Phacelia tanacetifolia Borkh.) honey composition was screened by UHPLC-DAD-QqTOF-MS. The targeted analysis revealed 6 major nitrogen compounds including aromatic amino acids (tyrosine, phenylalanine), purine derivatives (adenine, xanthine), nucleoside (uridine) and rare non-cyanogenic cyanoglucoside, (-)-5-epi-lithospermoside ((2Z)-2-[(4R,5R,6S)-4,5-dihydroxy-6-(ß-d-glucopyranosyl)oxycyclohex-2-en-1-ylidene]acetonitrile). Their identity was confirmed by different analytical tools: HRMS, co-chromatography with standard compound or comprehensive NMR experiments. All the compounds, except amino acids, were reported and determined in honey for the first time. The amount of the compounds was quantified in 16 unifloral phacelia samples: adenine (18.45 ±â€¯4.63 mg/kg), xanthine (10.53 ±â€¯2.98 mg/kg), uridine (42.84 ±â€¯9.26 mg/kg), tyrosine (14.66 ±â€¯10.22 mg/kg), (-)-5-epi-lithospermoside (70.61 ±â€¯31.37 mg/kg) and phenylalanine (20.41 ±â€¯11.99 mg/kg). The (-)-5-epi-lithospermoside content is significantly correlated with P. tanacetifolia pollen percentage (R2 = 0.5612, p < 0.001) and it is proposed as a potential marker of botanical origin for phacelia honey.

Occurrence of brominated disinfection byproducts in the air and water of chlorinated seawater swimming pools.

An undesirable consequence of disinfection is the formation of chemical contaminants known as disinfection byproducts (DBPs). Chronic exposure to DBPs has been linked to adverse health effects. The occurrence of DBPs in chlorinated pools filled with seawater (such as thalassotherapy pools and pools in spas) has received little attention so far. The present study evaluated the speciation and levels of disinfection byproducts in indoor swimming pools filled with seawater and treated with chlorine. Water and air samples were collected from three indoor swimming pools located in Southern France. Several classes of DBPs including trihalomethanes, haloacetic acids, haloacetonitriles, and trihaloacetaldehydes were analyzed in water. Halogenated volatile organic compounds were analyzed in air. Extractable organic halides (EOX) contents were determined using combustion/micro-coulometry system. The speciation of DBPs identified in the three pools was predominantly brominated. The mean (arithmetic) concentration of bromoform, dibromoacetic acid, tribromoacetic acid, dibromoacetonitrile and bromal hydrate in the three pools was 79.2, 72.9, 59.9, 26.9 and 10.0µg/L, respectively. By weight, HAAs represented the most abundant chemical class followed by THMs. In air, bromoform was the most abundant THM occurring at a mean concentration of 133.2µg/m3 in the three pools. The mean EOX level was 706µgCl-/L for the three pools. In average, the quantified DBPs accounted for only 14% of EOX, thus 86% of EOX remained unknown. Further research is warranted to identify the unknown DBPs.

Optimization of a one-step method for the multiresidue determination of organophosphorous pesticides in camellia oil.

UNLABELLED: Due to the widespread use and potential toxicity of organophosphorous pesticides (OPs), multiresidue monitoring of OPs in camellia oil has become increasingly important. A simple, rapid, and effective matrix solid-phase dispersion extraction for the determination of 15 organophosphorous pesticides in camellia oil is described. Related important factors influencing the extraction efficiency, such as type of sorbent material, eluting solvent, and ratio of sample/sorbent were studied and optimized. The best results were obtained using 0.5 g of camellia oil, 1.5 g of white carbon black as dispersant sorbent, and 5 mL of acetonitrile: ethyl acetate (2:1, V/V) as eluting solvent. Method validation was performed in order to study sensitivity, linearity, precision, and accuracy. Average recoveries ranged between 76.7% and 102.9% with relative standard deviation values from 2.9% to 13.7% at 2 concentration levels (10 and 100 µg/kg). The method limit of detection at or below the regulatory maximum residue limits for the pesticides was achieved. PRACTICAL APPLICATION: A simple, rapid, and effective method for multiresidue determination of organophosphorous pesticides in camellia oil was developed. The sample preparation could finish in 5 min.

Optimization of micellar electrokinetic capillary chromatography method using central composite design for the analysis of components in Yangwei granule.

Central composite design (CCD), together with multiple linear regression, was successfully used to optimize the electrophoretic buffer system of micellar electrokinetic capillary chromatography (MEKC) for the determination of albiflorin, paeoniflorin, liquiritin, and glycyrrhizic acid in the traditional Chinese medicine (TCM) prescription, Yangwei granule. Concentrations of sodium deoxycholate (SDC) and borate, and proportions of ammonia, acetonitrile, and methanol were optimized. The total resolutions of peaks between the analytes and their adjacent peaks in real samples were integrated into the evaluation index of separation efficiency. The optimum electrophoretic buffer contained 80 mmol/L SDC, 20 mmol/L borate, 5% (v/v) methanol, 0.5% (v/v) ammonia, and 5% (v/v) acetonitrile. The correlation coefficients (R(2)) between the peak areas and the corresponding concentrations of analytes were greater than 0.9956. The limits of detection (LODs) (S/N=3) of the analytes were 0.97-4.00 µg/ml. The results indicate the superiority of CCD in optimizing the separation conditions of complex samples such as TCM prescriptions.

Enhancement of fermentation process in Pu-erh tea by tea-leaf extract.

Pu-erh tea is known as a fermented tea and longer storage enhances its flavor and taste. Recently, Aspergillus, Blastobotrys, and Streptomyces are found to play important roles in nutritional enhancement of Pu-erh tea by fermentation. Since water and temperature affect the microbial growth, we therefore explored the factors that might enhance the Pu-erh tea fermentation. The results showed that the addition of fresh tea-leaf extract (TLE) enhanced the withered tea fermentation (at 37 degrees C, 80 to 85% RH) as compared with the water only. Contents of statin, GABA, gallic acid, DPPH scavenging and polyphenol oxidase (PPO) activities were increased, whereas polyphenols and caffeine were decreased over 6 mo. TLE dose-dependently enhanced some of the qualities (that is, statin, PPO) of Pu-erh tea significantly as compared with the water only. The effect was related to the increase population of A. niger and A. carbonarius at 6 mo (from 7.6 +/- 1.2 x 10(1) and 3.2 +/- 1.3 x 10(1) to 3.1 +/- 1.2 x 10(6) and 2.4 +/- 1.1 x 10(5) colony forming units [CFU]/g, respectively). After drying process (90 degrees C, 30 min), the total microbial count from these samples returned to background level (3 +/- 0.5 x 10(2) CFU/g). None of ochratoxin and fumonisin, toxins from Aspergillus, was detected in the final products. The flavor and taste were also enhanced by treatment with TLE. The inoculation with S. cinereus Y11 with 2% TLE further enhanced these functional contents (about 2-fold increase of statin level) in the experimental Pu-erh tea. Therefore, this result may add a new process for Pu-erh tea manufacture.

Fingerprint analysis of the fruits of Cnidium monnieri extract by high-performance liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry.

A method incorporating high-performance liquid chromatography (HPLC) with electrospray ionization (ESI) and tandem mass spectrometry (MS), with parallel analysis by HPLC with UV detection using a diode-array detector (DAD) was developed for the qualitative characterization of coumarin and chromone constituents in the fruits of Cnidium monnieri. The chromatographic separations were performed on a Diamonsil C18 column (4.6 mm x 200 mm, 5 microm) with water with 50 mM ammonium acetate and 2% acetic acid (A) and acetonitrile (B) as the mobile phase. According to the characteristic UV spectra, the information of molecular weight and structure provided by ESI-MS/MS, 13 coumarin and 7 chromone components were detected and identified. This method is rapid and reliable for identification of the constituents in the complex herbal system, and the fragmentation patterns proposed could be extended to the similar compounds.

Determination of aristolochic acid in botanicals and dietary supplements by liquid chromatography with ultraviolet detection and by liquid chromatography/mass spectrometry: single laboratory validation confirmation.

The presence of aristolochic acid in some dietary supplements is a concern to regulators and consumers. A method has been developed, by initially using a reference method as a guide, during single laboratory validation (SLV) for the determination of aristolochic acid I, also known as aristolochic acid A, in botanical species and dietary supplements at concentrations of approximately 2 to 32 microg/g. Higher levels were determined by dilution to fit the standard curve. Through the SLV, the method was optimized for quantification by liquid chromatography with ultraviolet detection (LC-UV) and LC/mass spectrometry (MS) confirmation. The test samples were extracted with organic solvent and water, then injected on a reverse phase LC column. Quantification was achieved with linear regression using a laboratory automation system. The SLV study included systematically optimizing the LC-UV method with regard to test sample size, fine grinding of solids, and solvent extraction efficiency. These parameters were varied in increments (and in separate optimization studies), in order to ensure that each parameter was individually studied; the test results include corresponding tables of parameter variations. In addition, the chromatographic conditions were optimized with respect to injection volume and detection wavelength. Precision studies produced overall relative standard deviation values from 2.44 up to 8.26% for aristolochic acid I. Mean recoveries were between 100 and 103% at the 2 microg/g level, between 102 and 103% at the 10 microg/g level, and 104% at the 30 microg/g level.

A high performance liquid chromatography method for the simultaneous determination of arctiin, chlorogenic acid and glycyrrhizin in a Chinese proprietary medicine.

A high performance liquid chromatography (HPLC) method was developed for the simultaneous determination of arctiin, chlorogenic acid and glycyrrhizin in the tablets of a Chinese proprietary medicine named, "Yin Qiao Jie Du Pian". The analysis was performed by a reverse phase gradient elution, using an aqueous mobile phase (containing 0.4% acetic acid and 4.5% tetrahydrofuran) modified by acetonitrile and detection made simultaneously at three wavelengths. The method was validated for specificity, accuracy, precision and limits of detection and quantification. Tablets of seven commercial brands were analyzed and found to contain different amounts of the three bioactive markers. This raised the question of the quality and the efficacy of the products. The method developed can be used for the quality control of "Yin Qiao Jie Du" tablets.