Multi‐residue method for the analysis of pharmaceutical compounds in sewage sludge,compost and sediments by sonication‐assisted extraction and LC determination

A method for the simultaneous determination of 16 pharmaceutical compounds in three types of sewage sludge (primary, secondary and anaerobically digested dehydrated sludge), compost and sediment samples is described. Pharmaceutical compounds evaluated were nonsteroidal anti‐inflammatory drugs (acetaminophen, diclofenac, ibuprofen, ketoprofen, naproxen and salicylic acid), antibiotics (sulfamethoxazole and trimethoprim), an anti‐epileptic drug (carbamazepine), a β‐blocker (propranolol), a nervous stimulant (caffeine), estrogens (17α‐ethinylestradiol, 17β‐estradiol, estriol and estrone) and lipid regulators (clofibric acid, metabolite of clofibrate and gemfibrozil). The method is based on the ultrasonic‐assisted extraction, clean‐up by SPE and analytical determination by HPLC with diode array and fluorescence detectors. The best extraction recoveries were achieved in a three‐step extraction procedure with methanol and acetone as extraction solvents. Extraction recoveries of several pharmaceutical compounds as caffeine were highly dependent on the type of sample evaluated. The applicability of the method was tested by analyzing primary, secondary and anaerobically digested dehydrated sludge, compost and sediment samples from Seville (Southern Spain). Ten of the sixteen pharmaceutical compounds were detected in sludge samples and five in compost and sediment samples. The highest concentration levels were recorded for ibuprofen in sewage samples, whereas salicylic acid and 17α‐ethinylestradiol were detected in all of the samples analyzed.     

The use of molecularly imprinted polymers for the multicomponent determination of endocrine‐disrupting compounds in water and sediment

The method employing molecularly imprinted polymers for the extraction and clean up of endocrine‐disrupting compounds (estrogens, bisphenol A, and alkylphenols) from water and sediment is described. The identical extraction/clean‐up and LC‐MS/MS condition were used for the analysis of both types of samples. The method showed high recoveries ranging from 90 to 99% with excellent precision (intrabatch: 3.6–9.3%; interbatch: 5.6–11.4% for water; intrabatch: 4.3–8.5%; interbatch: 6.1–9.6% for sediment). The LOD was in the range of 0.7–1.9 ng/L and 0.3–0.6 ng/g for water and sediment, respectively. Overall extraction on molecularly imprinted polymers substantially enhanced sample clean‐up. The difference in efficiency of clean‐up was particularly pronounced when a large sample volume/weight was extracted and analyzed. Finally, the method was successfully applied for the analysis of 20 water and sediment samples.     

超高效液相色谱-串联质谱法同时检测地表水中18种药物与个人护理品的残留量

采用固相萃取对水样进行预处理,建立了同时检测地表水中包括抗生素、β-阻滞剂、驱蚊剂、抗癫痫药、中枢神经兴奋剂、血脂调节剂、非甾体抗炎药、杀菌消毒剂在内的18种药物与个人护理品的超高效液相色谱-串联质谱分析方法。采用中性条件萃取水样,控制上样流速为2 mL/min,用甲醇-乙腈(1:1, v/v)溶液洗脱。纯水中的平均加标回收率为53.9%～112%,相对标准偏差为2.6%～15.3%(n=6);以地表水样加标100 ng/L为样品,目标分析物平均回收率为45.1%～156.6%,相对标准偏差为2.4%～15.7%(n=6)。结果表明,本方法可同时精确检测地表水样中的18种分析物,方法验证结果表明所建立的方法可靠。用该方法分析杭州余杭塘河水,结果检出9种分析物,其中咖啡因平均质量浓度达550.7 ng/L。结果表明该方法可靠。     

Simultaneous determination of triclosan,triclocarban, and transformation products of triclocarban in aqueous samples using solid‐phase micro‐extraction‐HPLC‐MS/MS

The presence of triclosan and triclocarban, two endocrine‐disrupting chemicals and antimicrobial agents, and transformation products of triclocarban, 1,3‐di(phenyl)urea, 1,3‐bis(4‐chlorophenyl)urea and 1,3‐bis(3,4‐dichlorophenyl)urea, in tap water, treated household drinking water, bottled water, and river water samples were investigated using solid‐phase micro‐extraction coupled with‐HPLC‐MS/MS, a rapid, green, and sensitive method. Factors influencing the quantity of the analytes extracted onto the solid‐phase micro‐extraction fiber, such as addition of salt, sample pH, extraction time, desorption time, and sample volume, were optimized using solid‐phase micro‐extraction‐HPLC‐MS/MS. The results showed that the method gave satisfactory sensitivities and precisions for analyzing sub‐part‐per‐trillion levels of triclosan, triclocarban, and transformation products of triclocarban in samples collected locally. The recoveries of analytes ranged from 97 to 107% for deionized water samples, and 99 to 110% for river water samples, and limits of detection were in the range of 0.32–3.44 and 0.38–4.67 ng/L for deionized water and river water samples, respectively. On average, the daily consumption of triclosan and triclocarban by an adult by consuming 2 liters of different types of drinking water were estimated to be in the range of 6.13–425 ng/day as a result of the concentrations of triclosan and triclocarban measured in this study.     

Speciation analysis of mercury in sediments using ionic‐liquid‐based vortex‐assisted liquid–liquid microextraction combined with high‐performance liquid chromatography and cold vapor atomic fluorescence spectrometry

An improved novel method based on ionic liquid vortex‐assisted liquid–liquid microextraction has been developed for the extraction of methylmercury, ethylmercury and inorganic mercury in sediment samples prior to analysis by high‐performance liquid chromatography with cold vapor atomic fluorescence spectrometry. In this work, mercury species were firstly complexed with dithizone, and the complexes were extracted into 1‐hexyl‐3‐methylimidazolium hexafluorophosphate. Key factors that affect the extraction efficiency of mercury species, such as type and amount of ionic liquid and chelatants, extraction time, sample pH, salt effect and matrix effect were investigated. Under the optimum conditions, linearity was found in the concentration range from 0.1–70 ng/g. Limits of detection ranged from 0.037–0.061 ng/g. Reproducibility and recoveries were assessed by extracting a series of six independent sediment samples that were spiked with different concentration levels. Finally, the proposed method was successfully applied in analysis of real sediment samples. In this work, ionic liquids vortex‐assisted liquid–liquid microextraction was for the first time used for the extraction of mercury species in sediment samples. The proposed method was proved to be much simpler and more rapid, as well as more environmentally friendly and efficient compared with the previous methods.     

Metal–organic framework UiO‐67‐coated fiber for the solid‐phase microextraction of nitrobenzene compounds from water

A sol–gel coating technique was applied for the preparation of a solid‐phase microextraction fiber by coating the metal–organic framework UiO‐67 onto a stainless‐steel wire. The prepared fiber was explored for the headspace solid‐phase microextraction of five nitrobenzene compounds from water samples before gas chromatography with mass spectrometric detection. The effects of the extraction temperature, extraction time, sample solution volume, salt addition, and desorption conditions on the extraction efficiency were optimized. Under the optimal conditions, the linearity was observed in the range of 0.015–12.0 μg/L for the compounds in water samples, with the correlation coefficients (r) of 0.9945–0.9987. The limits of detection of the method were 5.0–10.0 ng/L, and the recoveries of the analytes from spiked water samples for the method were in the range of 74.0–102.0%. The precision for the measurements, expressed as the relative standard deviation, was less than 11.9%.     

Simultaneous determination of pharmaceuticals, endocrine disrupting compounds and hormone in soils by gas chromatography-mass spectrometry

Analytical methods have been developed for simultaneous determination of six different pharmaceuticals and personal care products (PPCPs) (clofibric acid, ibuprofen, naproxen, ketoprofen, diclofenac, and triclosan), three endocrine disrupting compounds (EDCs) (4-tert-octylphenol, 4-n-nonylphenol, and bisphenol A (BPA)) and one estrogenic compound (estrone) in soil matrix. The soils were extracted by different solvents with the help of an ultrasonic treatment at 42 kHz, followed by a solid phase extraction (SPE) as a cleanup procedure. The purified extracts were derivatized with N-methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA) and then analyzed by GC-MSD (SIM mode). The method was evaluated by testing the following variables: initial spiking levels, extraction solvents, solvent volumes, and soil types (sandy and clay soils). For 5 g of soil, four successive extraction steps with the mixture of acetone-ethyl acetate provided satisfactory recoveries. In the sandy soil, the recoveries of all the compounds were from 63.8 to 110.7% for the spiking level of 100 ng/g dry soil, and from 52.2 to 108.2% for 5 ng/g dry soil, respectively. Result was similar for the clay soil. The precision across all recoveries was high, suggesting that this method has a good reproducibility. The method was successfully employed to soil samples collected from a golf course irrigated with reclaimed wastewater in southern California, and resulted in the detection of clofibric acid, ibuprofen, naproxen, triclosan, bisphenol A, and estrone at ng per gram dry weight concentration levels. The method is robust and simple, and provides straightforward analyses of these current-emerging trace organic pollutants in solid matrices.     

Preparation of dummy‐imprinted polymers by Pickering emulsion polymerization for the selective determination of seven bisphenols from sediment samples

The dummy molecularly imprinted polymers were prepared by Pickering emulsion polymerization. 4,4′‐(1‐Phenylethylidene) bisphenol was selected as the dummy template to avoid the leakage of the target bisphenols. The microsphere particles were characterized by scanning electron microscopy and nitrogen adsorption–desorption measurements, demonstrating that the regular‐shaped and medium‐sized particles (40–70 μm) were obtained with a specific surface area of 355.759 m²/g and a total pore volume of 0.561 cm³/g. The molecular imprinting properties of the particles were evaluated by static adsorption and chromatographic evaluation experiments. The association constant and maximum adsorption amount of bisphenol A were 0.115 mmol/L and 3.327 μmol/g using Scatchard analysis. The microsphere particles were then used as a solid‐phase extraction sorbent for selective extraction of seven bisphenols. The method of dummy molecularly imprinted solid‐phase extraction coupled with high‐performance liquid chromatography and diode array detection was successfully established for the extraction and determination of seven bisphenols from environmental sediment samples with method detection limits of 0.6–1.1 ng/g. Good recoveries (75.5–105.2%) for sediment samples at two spiking levels (500 and 250 ng/g) and reproducibility (RSDs < 7.7%, n = 3) were obtained.     

Vortex‐assisted extraction combined with dispersive liquid–liquid microextraction for the determination of polycyclic aromatic hydrocarbons in sediment by high performance liquid chromatography

A simple, rapid, and efficient method, vortex‐assisted extraction followed by dispersive liquid–liquid microextraction (DLLME) has been developed for the extraction of polycyclic aromatic hydrocarbons (PAHs) in sediment samples prior to analysis by high performance liquid chromatography fluorescence detection. Acetonitrile was used as collecting solvent for the extraction of PAHs from sediment by vortex‐assisted extraction. In DLLME, PAHs were rapidly transferred from acetonitrile to dichloromethane. Under the optimum conditions, the method yields a linear calibration curve in the concentration range from 10 to 2100 ng g^?1 for fluorene, anthracene, chrysene, benzo[k]fluoranthene, and benzo[a]pyrene, and 20 to 2100 ng g^?1 for other target analytes. Coefficients of determinations ranged from 0.9986 to 0.9994. The limits of detection, based on signal‐to‐noise ratio of three, ranged from 2.3 to 6.8 ng g^?1. Reproducibility and recoveries was assessed by extracting a series of six independent sediment samples, which were spiked with different concentration levels. Finally, the proposed method was successfully applied in analyses of real nature sediment samples. The proposed method extended and improved the application of DLLME to solid samples, which greatly shorten the extraction time and simplified the extraction process.     

Magnetic solid‐phase extraction of benzoylurea insecticides by Fe3O4 nanoparticles decorated with a hyper‐cross‐linked porous organic polymer

A magnetic polytriphenylamine porous organic polymer was prepared through simple self‐polycondensation of triphenylamine followed by coprecipitation with Fe²⁺ and Fe³⁺. It was applied as a magnetic adsorbent for the extraction of six benzoylurea insecticides from tomato, cucumber, and watermelon samples before their high‐performance liquid chromatography and mass spectral detection. Under the optimized experimental conditions, the established method gave a low limit of detection ranging from 0.05 to 0.1 ng/g and a good linear response ranging from 0.2 to 40 ng/g with coefficients of determination >0.99. The method recoveries for spiked analytes at the concentrations of 3 and 15 ng/g in real samples were in the range of 87.7–106.7% with the relative standard deviations <6.4%. The results indicated that it had a good adsorption capability toward the target analytes due to the π‐stacking and hydrogen bonding interactions. The polymer material showed great potential in the efficient extraction of organic compounds from real samples with complex matrixes.     

Extraction and analysis of pharmaceuticals in polluted sediment using liquid chromatography mass spectrometry

A multi-residue method for the extraction and clean-up of sediment samples was developed for the analysis of pharmaceutical residues. Sediment samples were collected in the proximity of sewage water plant in Stockholm, Sweden. Target analytes were the basic β-blocker propranolol, the neutral neuroleptic carbamazepine and the acidic anticoagulant warfarin, the painkiller diclofenac and the lipid regulator gemfibrozil. The extraction solvent was optimised with regard to pH and organic modifer. Extraction and clean up were performed with liquid-liquid extraction and ultra-sonication followed by solid-phase extraction. One extraction solvent, containing acetone/McIlvaine buffer pH4, provided satisfactory extraction for all substances. LC/MSMS in the MRM mode was used for determination. The recoveries of the extraction and clean-up steps were 60–75% (±2–8%) and LOQs were in the range 0.4–8?ng/g sediment (dry weight). The pharmaceuticals found in the sediment samples were propranolol and carbamazepine, representing substances with basic and neutral properties. Additionally, the samples were analysed with LC/QTOF for verification with the use of accurate mass measurement in the full-scan mode. Pharmaceuticals not represented in the original method were looked for. Non-target pharmaceuticals found using the LC/QTOF system were the basic β-blocker metoprolol and the acidic painkiller naproxen.     

Microwave-assisted solvent extraction of solid matrices and subsequent detection of pharmaceuticals and personal care products (PPCPs) using gas chromatography-mass spectrometry

Concentrations of pharmaceuticals and personal care products (PPCPs) in natural solids remain largely unknown. Contributing to this, is a lack of methods permitting the simultaneous detection of the diverse, low-level contaminants present in these complex matrices. We have developed a microwave-assisted solvent extraction (MASE)-based method targeting seven diverse PPCPs (caffeine, 17β-estradiol, ibuprofen, ketoprofen, musk ketone, naproxen, and triclosan) and a molecular marker for fecal waste (epicoprostanol). The method consisted of optimizing the following variables: derivatization of the polar target analytes, silica gel open column clean-up, and gas chromatographic-mass spectrometric (GC-MS) analysis of sample extracts for analysis and detection of the compounds noted above. Testing of the method on spiked soil allowed for 89.6 ± 2.89% recovery of three target compounds and 25.0 ± 1.93% recovery of five of the compounds. Although the latter recoveries were low, the precision across all recoveries was high, suggesting good reproducibility in application of the method. Furthermore, we suspect that matrix effects are likely responsible for the lower recoveries. Techniques with the exclusive incorporation of organic solvents were found inapplicable in the study of a pharmaceutical salt, diphenhydramine HCl. Application of the developed method to sediment collected directly downstream of the effluent pipe of a wastewater treatment plant allowed detection of ibuprofen, naproxen, ketoprofen, and epicoprostanol at ng-μg per gram dry weight concentrations. The observation of acidic pharmaceuticals, previously believed to exhibit insignificant sorption to solid matrices, in the tested sediment samples, coupled with application of biosolids for agricultural purposes, demonstrates the need for expanded investigation of PPCP contamination of natural solid matrices.     

Determination of five sulfonylurea herbicides in environmental waters and soil by ultra high performance liquid chromatography with tandem mass spectrometry after extraction using graphene

A fast and novel analytical method was developed for the determination of trace levels of sulfonylurea herbicides in water and soil samples. Graphene was used as a sorbent for extraction, and ultra high performance liquid chromatography with tandem mass spectrometry was used for quantification. Five sulfonylurea herbicides were preconcentrated from water samples using a graphene‐loaded packed cartridge, while extraction from soil samples was performed in a single step using graphene‐supported matrix solid‐phase dispersion. Under the optimized conditions, the calibration plots were linear in the range between 5 and 1000 ng/L for water samples, and between 1 and 200 ng/g for soil samples. All correlation coefficients (R) were >0.99. The limits of detection for water and soil samples were 0.28–0.53 ng/L and 0.08–0.26 ng/g, respectively. This method was successfully applied to the analysis of spiked samples of environmental water and soil, with recoveries ranging from 84.2–109.3 and 86.12–103.2%, respectively, all with relative standard deviations of <10%.     

Fast ultrasound‐assisted extraction combined with LC–MS/MS of perfluorinated compounds in manure

A fast analytical method for the determination of perfluorinated compounds in poultry manure by LC–MS/MS was developed. The extraction was carried out by ultrasound‐assisted extraction of 1 g of sample, during 2 × 15 min using low volume (5.5 mL) of a mixture of methanol and acetonitrile. An efficient extraction of perfluoroalkyl carboxylates, perfluoroalkyl sulfonates, and perfluoroalkyl sulfonamides from poultry manure was obtained with recoveries higher than 81%. The cleanup of extracts was carried out by dispersive SPE. The validation of the proposed method showed the suitability of this procedure to determine perfluorinated compounds in poultry manure with detection limits in the range of 0.44–2.12 ng/g, depending on the target compound. In comparison with previously published methods, the miniaturization of the sample preparation method with ultrasound‐assisted extraction together with the use of a core‐shell column permit a lower consumption of organic solvents and a fast analysis of perfluorinated compounds. Manure samples obtained from Spanish commercial farms were analyzed and low perfluorinated compounds levels were found, which may be originated by dietary or environmental exposure. The highest concentrations measured corresponded to the perfluoroalkyl sulfonates, which varied from 8.2 to 35.9 ng/g.     

Determination of trace triclosan in environmental water by microporous bamboo‐activated charcoal solid‐phase extraction combined with HPLC‐ESI‐MS

A sensitive and efficient analytical method for triclosan (TCS) determination in water, which involves enrichment with bamboo‐activated charcoal and detection with HPLC‐ESI‐MS, was developed. The influence of several operational parameters, including the eluant and its volume, the flow rate, the volume andacidity of the sample, and the amount of bamboo‐activated charcoal, were investigated and optimized. Under the optimum conditions, linearity of the method was observed in the range of 0.02–20 μg/L, with correlation coefficients (r²) >0.9990. The limit of detection was 0.002 μg/L based on the ratio of chromatographic signal to baseline noise (S/N = 3). The spiked recoveries of TCS in real water samples were achieved in the range of 97.6–112.5%. The proposed method was applied to analyze TCS in real aqueous samples. All the surface water samples collected in Xiaoqing River had detectable levels of TCS with concentrations of 42–197 ng/L.     

Fabric phase sorptive extraction/GC‐MS method for rapid determination of broad polarity spectrum multi‐class emerging pollutants in various aqueous samples

A rapid extraction and cleanup method using selective fabric phase sorptive extraction combined with gas chromatography and mass spectrometry has been developed and validated for the determination of broad polarity spectrum emerging pollutants, ethyl paraben, butyl paraben, diethyl phthalate, dibutyl phthalate, lidocaine, prilocaine, triclosan, and bisphenol A in various aqueous samples. Some important parameters of fabric phase sorptive extraction such as extraction time, matrix pH, stirring speed, type and volume of desorption solvent were investigated and optimized. Calibration curves were obtained in the concentration range 0.05–500 ng/mL. Under the optimum conditions, the limits of detection were in the range 0.009 –0.021 ng/mL. This method was validated by analyzing the compounds in spiked aqueous samples at different levels with recoveries of 93 to 99% and relative standard deviations of <6%. The developed method was applied for the determination of the emerging contaminants in tap water, municipal water, ground water, sewage water, and sludge water samples. The results demonstrate that fabric phase sorptive extraction has great potential in the preconcentration of trace analytes in complex matrix.     

Microwave‐assisted extraction combined with on‐line solid phase extraction followed by ultra‐high‐performance liquid chromatography with tandem mass spectrometric determination of benzotriazole UV stabilizers in marine sediments and sewage sludges

Benzotriazole ultra‐violet stabilisers are compounds widely used in personal care products, which can reach the environment after passing through wastewater treatment plants. In this work, we develop a novel method to evaluate the presence of seven compounds in marine sediments and sewage sludges using microwave‐assisted extraction followed by a clean‐up step based in on‐line solid phase extraction coupled to ultra‐high‐performance liquid chromatography with MS/MS detection. This method allows for fast and efficient extraction from the solid matrix, subsequent automatic on‐line purification and preconcentration, and analysis. For the optimised method, LOD were from 53.3 to 146 ng/kg and LOQ were in the range of 176–486 ng/kg. The method was validated for different environmental solid samples with satisfactory recoveries and relative standard deviations, between 46.1 and 83.9 and 7.8 and 15.5% (sludges) and 50.1 and 87.1% and 8.83 and 16.3% (sediments), respectively. Finally, the studied analytes were quantified in concentrations between 0.18 and 24.0 ng/g in real samples of marine sediments and sewage sludges from Gran Canaria Island (Spain).     

A simple solvent collection technique for a dispersive liquid–liquid microextraction of parabens from aqueous samples using low‐density organic solvent

A simple technique for the collection of an extraction solvent lighter than water after dispersive liquid–liquid microextraction combined with high‐performance liquid chromatography with ultraviolet detection was developed for the determination of four paraben preservatives in aqueous samples. After the extraction procedure, low‐density organic solvent together with some little aqueous phase was separated by using a disposable glass Pasteur pipette. Next, the flow of the aqueous phase was stopped by successive dipping the capillary tip of the pipette into anhydrous Na₂SO₄. The upper organic layer was then removed simply with a microsyringe and injected into the high‐performance liquid chromatography system. Experimental parameters that affect the extraction efficiency were investigated and optimized. Under optimal extraction conditions, the extraction recoveries ranged from 25 to 86%. Good linearity with coefficients with the square of correlation coefficients ranging from 0.9984 to 0.9998 was observed in the concentration range of 0.001–0.5 μg/mL. The relative standard deviations ranged from 4.1 to 9.3% (n = 5) for all compounds. The limits of detection ranged from 0.021 to 0.046 ng/mL. The method was successfully applied for the determination of parabens in tap water and fruit juice samples and good recoveries (61–108%) were achieved for spiked samples.     

Development of pressurized liquid extraction and solid‐phase microextraction combined with gas chromatography and flame photometric detection for the determination of organophosphate esters in sediments

Organophosphate esters have been extensively used as flame retardants and plasticizers. The analysis of organophosphate esters in the environment is a hot topic because many of them are toxic and persistent. We developed a novel procedure for determining organophosphate esters in sediment. In this work, pressurized liquid extraction and solid‐phase microextraction are used for sample preparation to extract and concentrate the analytes, which are then analyzed by gas chromatography with flame photometric detection. The extraction parameters of pressurized liquid extraction were investigated and optimized by orthogonal design and then evaluated by range analysis and analysis of variance. Under the optimal conditions, the proposed procedure showed wide linear ranges (0.90–100 ng/g) with correlation coefficients ranging from 0.9921 to 0.9990. The detection limits of the method were in the range of 0.009–0.280 ng/g with standard deviations ranging from 2.2 to 9.5%. Recoveries of the proposed method ranged from 82.3 to 108.9% with relative standard deviations <8.4%. The obtained method was applied successfully to the determination of organophosphate esters in real sediments with recoveries varying from 79.8 to 107.3%. The proposed method was proved to be simple, easy, and sensitive for analyzing organophosphate esters in sediment samples.     

Application of mesoporous carbon as a solid‐phase microextraction fiber coating for the extraction of volatile aromatic compounds

A mesoporous carbon was fabricated using MCM‐41 as a template and sucrose as a carbon source. The carbon material was coated on stainless‐steel wires by using the sol–gel technique. The prepared solid‐phase microextraction fiber was used for the extraction of five volatile aromatic compounds (chlorobenzene, ethylbenzene, o‐xylene, bromobenzene, and 4‐chlorotoluene) from tea beverage samples (red tea and green tea) prior to gas chromatography with mass spectrometric detection. The main experimental parameters affecting the extraction of the volatile aromatic compounds by the fiber, including the extraction time, sample volume, extraction temperature, salt addition, and desorption conditions, were investigated. The linearity was observed in the range from 0.1 to 10.0 μg/L with the correlation coefficients (r) ranging from 0.9923 to 0.9982 and the limits of detection were less than 10.0 ng/L. The recoveries of the volatile aromatic compounds by the method from tea beverage samples at spiking levels of 1.0 and 10.0 μg/L ranged from 73.1 to 99.1%.