Unknown residual solvents identification in drug products by headspace solid phase microextraction gas chromatography-mass spectrometry

Summary A sensitive headspace SPME method for the extraction of residual solvents from pharmaceutical products has been developed and optimized. It was found that minimizing sample and headspace volume has a beneficial effect on extraction efficiency. At the same time the method reproducibility was seriously affected by reducing sample and headspace volume. The added air volume was not found to have any significant influence on method sensitivity. The method showed reproducibilities of less than 10% and detection limits as low as 1 ppb for benzene and dichloromethane. The headspace SPME method is around 1000 times more sensitive than static headspace. The optimized parameters were headspace volume 1.5 mL, sample volume 10 μL, and extraction time 30 min. The method was successfully applied to the identification of unknown residual solvents in three different proprietary active drug substances and was successfully applied to the confirmation of the presence of benzene in a proprietary drug substance. Presented at Balaton Symposium '01 on High-Performance Separation Methods, Siófok, Hungary, September 2–4, 2001     

Monitoring of PAHs in air by collection on XAD-2 adsorbent then microwave-assisted thermal desorption coupled with headspace solid-phase microextraction and gas chromatography with mass spectrometric detection

Microwave-assisted thermal desorption (MAD) coupled to headspace solid-phase microextraction (HS-SPME) has been studied for in-situ, one-step, sample preparation for PAHs collected on XAD-2 adsorbent, before gas chromatography with mass spectrometric detection. The PAHs on XAD-2 were desorbed into the extraction solution, evaporated into the headspace by use of microwave irradiation, and absorbed directly on a solid-phase microextraction fiber in the headspace. After desorption from the SPME fiber in the hot GC injection port, PAHs were analyzed by GC–MS. Conditions affecting extraction efficiency, for example extraction solution, addition of salt, stirring speed, SPME fiber coating, sampling temperature, microwave power and irradiation time, and desorption conditions were investigated. Experimental results indicated that extraction of 275 mg XAD-2, containing 10–200 ng PAHs, with 10-mL ethylene glycol–1 mol L⁻¹ NaCl solution, 7:3, by irradiation with 120 W for 40 min (the same as the extraction time), and collection with a PDMS–DVB fiber at 35 °C, resulted in the best extraction efficiency. Recovery was more than 80% and RSD was less than 14%. Optimum desorption was achieved by heating at 290 °C for 5 min. Detection limits varied from 0.02 to 1.0 ng for different PAHs. A real sample was obtained by using XAD-2 to collect smoke from indoor burning of joss sticks. The amounts of PAHs measured varied from 0.795 to 2.53 ng. The method is a simple and rapid procedure for determination of PAHs on XAD-2 absorbent, and is free from toxic organic solvents.     

Multiple headspace single-drop microextraction coupled with gas chromatography for direct determination of residual solvents in solid drug product

This paper proposed a multiple headspace single-drop microextraction (MHS-SDME) method coupled to gas chromatography with flame-ionization detection (GC-FID) for direct determination of residual solvents in solid drug product. The MHS-SDME technique is based on extrapolation to an exhaustive extraction of consecutive extractions from the same sample which eliminates the matrix effect on the quantitative analysis of solid samples. The total peak area of analyte is calculated with a beta constant which can be obtained from the slope of the linear regression that related to the peak area of each extraction and the number of extraction times. In this work, a model drug powder was chosen and the amounts of residues of two solvents, methanol and ethanol, were investigated. The factors influencing the extraction process including extraction solvent, microdrop volume, extraction time, sample amount, thermostatting temperature and incubation time were studied. 10 mg of drug powder was incubated for 3 h at 140 °C prior to the first extraction and thermostatted for 15 min at 140 °C between each extraction. Extraction was carried out with 2 μL of dimethyl sulfoxide (DMSO) as the microdrop for 5 min. The features of the method were established using standard solutions. Validation of the proposed method showed good agreement with the traditional dissolution method for analysis of residual solvents in drug product. The results indicated that MHS-SDME has a great potential for the quantitative determination of residual solvents directly from the solid drug products due to its low cost, ease of operation, sensitivity, reliability and environmental protection.     

Headspace sorptive extraction using silicone tubes for the determination of chlorobenzenes in water

A new approach for headspace sorptive extraction is presented and demonstrated for the determination of 12 chlorobenzenes in water samples. It consists of a silicone tube (15-mm length) arranged around a stainless steel rod. This device is fixed on a septum cap and exposed to the headspace of 50 mL of a salt-saturated water sample. After extraction (60-min optimized extraction time), thermodesorption is carried out by direct insertion of the silicone tube into the thermodesorption-gas chromatography-mass spectrometry system. Desorption of the analytes is performed at 250 °C for 5 min with a gas flow of 50 mL/min. Repeatability (relative standard deviation 5–10%), extraction yields (9–46%), enrichment factors (129–657), and detection limits (0.002–0.012 μg/L) were determined and four real water samples were analyzed with the headspace tube extraction. The results were verified by standard addition. A comparison of headspace tube extraction with other headspace enrichment techniques underlined the high extraction capacity of the proposed method. A big advantage of tube extraction is the low cost of the silicone material. The tubes can be discarded after single use, avoiding carryover problems and cross-contamination. Figure Scheme of the HS-tube extraction and thermodesorption system     

Determination of different recreational drugs in hair by HS-SPME and GC/MS

A simple procedure combining headspace solid-phase microextraction (HS-SPME) and gas chromatography–mass spectrometry (GC/MS) to detect and quantify amphetamines, ketamine, methadone, cocaine, cocaethylene and ∆⁹-tetrahydrocannabinol (THC) in hair is described. This procedure allows, in a single sample, even scant, analysis of drugs requiring different analytical conditions. A hair sample (10 mg) is washed and subjected to acidic hydrolysis. Then the HS-SPME is carried out (10 min at 90 °C) for amphetamines, ketamine, methadone, cocaine and cocaethylene. For derivatization of analytes, the fibre is introduced into the headspace of another closed vial containing acetic anhydride. After a chromatographic run, an alkaline hydrolysis for THC analysis is carried out in the same vial containing the hair sample previously used. For adsorption, the solid-phase microextraction needle is inserted into the headspace of the vial and the fibre is exposed for 30 min at 150 °C. For derivatization of analytes, the fibre is introduced into the headspace of another closed vial containing N-methyl-N-(trimethylsilyl)trifluoroacetamide. The GC/MS parameters were the same for both chromatographic runs. The linearity was proved to be between 0.01 and 10.00 ng/mg. The repeatability (intra- and interday precision) was below 10% as the coefficient of variation for all compounds. The accuracy, as the relative recovery, was 96.2–103.5% (spiked samples) and 88.6–101.7% (quality control sample). The limit of detection ranged from 0.01 to 0.12 ng/mg, and the limit of quantification ranged from 0.02 to 0.37 ng/mg. Application of the procedure to real hair samples is described. To the best of our knowledge, the proposed procedure combining HS-SPME and GC/MS is the first one be to successfully applied to the simultaneous determination of most of the common recreational drugs, including THC, in a single hair sample.     

New cloud vapor zone (CVZ) coupled headspace solid-phase microextraction technique

A new cloud vapor zone (CVZ)-based headspace solid-phase microextraction (HS-SPME) technique has been demonstrated with the capability of heating the sample matrix and simultaneously cooling the sampling zone. A bi-temperature-controlled (BTC) system, allowing 10 mL of test sample heating and headspace external-cooling, was employed for the CVZ formation around the SPME-fiber sampling area. In the CVZ procedure, the heated headspace vapor undergoes a sudden cooling near the SPME to form a dense cloud of analyte–water vapor, which is helpful for adsorption or absorption of the analyte. The device was evaluated for the quantitative analysis of aqueous chlorothalonil. Parameters influencing sampling efficiency, e.g., SPME fiber coating, SPME sampling temperature and time, solution modifier, addition of salt, sample pH, and temperature, were investigated and optimized thoroughly. The proposed BTC-HS-SPME method afforded a best extraction efficiency of above 94% accuracy (less than 4.1% RSD, n = 7) by using the PDMS fiber to collect chlorothalonil in the headspace at 5 °C under the optimized condition, i.e., heating sample solution (added as 10% ethylene glycol and 30% NaCl, at pH 7.0) at 130 °C for 15 min. The detection was linear from 0.01 to 80 μg L⁻¹ with a regression coefficient of 0.9998 and had a detection limit of 3.0 ng L⁻¹ based on S/N = 3. Practical application was demonstrated by analyzing chlorothalonil in farm water samples with promising results and recoveries. The approach provided a very simple, fast, sensitive, and solvent-free procedure to collect analytes from aqueous solution. The approach can provide a new platform for other sensitive HS-SPME assays.     

Determination of triethanolamine in air samples by gas chromatography-mass spectrometry

Summary Low amounts of triethanolamine, collected in ORBO 53 tubes during air sampling, required the development of a very sensitive method for determination. After desorption and silylation reaction with trimethylsilyl imidazole/trimethyl chlorosilane, the derivative was analyzed by gas chromatography-mass spectrometry on an Ultra 2 silica capillary column in single ion monitoring mode (retention time: about 6 min). The method has a detection limit of 1–2 pg with a desorption efficiency of about 81%. Linearity of response was ascertained in the ranges 10–100 ng and 100–1000 ng. Short-term method validation was carried out by intra- and inter-day assays on three amounts for each reference calibration curve. All results satisfied the pre-defined acceptance criteria. In general, the whole procedure was easily performed and was appropriate for our needs. Breakthrough volume was appropriate for our needs. Breakthrough volume was determined on authentic samples and was about 40–60 L, using a flow rate of 1 L·min⁻¹. The amounts of triethanolamine found in the samples ranged from 150 to 250 ng (about 2.5–4.2 μg·m⁻³).     

Determination of Mn, Fe, and Cu in chemically-treated wood pulps by the XRF addition method

A rapid X-ray fluorescence addition method has been developed for quantification of the technically most important metals in wood pulp matrix (Mn, Fe, and Cu). Pretreatment consisted of just two steps: first, acid was added to the sample to achieve homogeneous distribution of the metals; the pulp was then pressed lightly on to Mylar film. Total analysis time was less than 10 min. The concentration range investigated was up to 15 mg kg^–1 for Mn and up to 5 mg kg^–1 for Fe and Cu. Metal concentrations in Scandinavian pulps are not expected to exceed these amounts. The quantification limit was 2 mg kg^–1 for all three metals. The reproducibilities and repeatabilities were concentration-dependent and varied between 3 and 19% and between 1 and 17%, respectively. The squares of the linear correlation coefficients between measured intensity and added metal concentration were 0.994, 0.950, and 0.932 for Mn, Fe, and Cu, respectively. Received: 19 February 2001 / Revised: 17 April 2001 / Accepted: 19 April 2001     

A headspace solid-phase microextraction procedure coupled with gas chromatography–mass spectrometry for the analysis of volatile polycyclic aromatic hydrocarbons in milk samples

A sensitive and solvent-free method for the determination of ten polycyclic aromatic hydrocarbons, namely, naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene and chrysene, with up to four aromatic rings, in milk samples using headspace solid-phase microextraction and gas chromatography–mass spectrometry detection has been developed. A polydimethylsiloxane–divinylbenzene fiber was chosen and used at 75°C for 60 min. Detection limits ranging from 0.2 to 5 ng L⁻¹ were attained at a signal-to-noise ratio of 3, depending on the compound and the milk sample under analysis. The proposed method was applied to ten different milk samples and the presence of six of the analytes studied in a skimmed milk with vegetal fiber sample was confirmed. The reliability of the procedure was verified by analyzing two different certified reference materials and by recovery studies. Figure Milk is safe, healthy food     

Determination of volatile residual solvents in pharmaceutical products by headspace liquid-phase microextraction gas chromatography-flame ionization detector

This paper demonstrates headspace liquid-phase microextraction (HS-LPME) as used for the determination of volatile residual solvents in pharmaceutical products. This method is based on headspace liquid-phase microextraction capillary column gas chromatography. Under optimum conditions, the linerary of the method ranged from 1 to 1,000 mg l⁻¹. The limits of detection are 0.2–2.0 mg l⁻¹ and relative standard deviations (RSD) for most of the volatile solvents were below 10%. This novel method is applied to the analysis of volatile residual solvents in pharmaceutical products with satisfactory results.     

Optimisation and comparison of several microextraction/methylation methods for determining haloacetic acids in water using gas chromatography

This article presents the different modes and configurations of liquid-phase microextraction (LPME) through comparison with headspace solid-phase microextraction (HS-SPME) for the simultaneous extraction/methylation of the nine haloacetic acids (HAAs) found in water. This is the first analytical case reported of solvent bar extraction–preconcentration–derivatisation assisted by an ion-pairing transfer for HAAs. In this method, 5 μL of the organic extractant, decane, was confined within a hollow-fibre membrane that was placed in a stirred aqueous sample containing the derivatising reagents (dimethylsulphate with a tetrabutylammonium salt). With heating at 45 °C in the HS-SPME method, some organic solvents (extractant, excess of derivatising reagent) are also volatilised and compete with the esters on the fibre (the fibre is damaged and it can be reused only 50−60 times). In addition, the HS-SPME method provides inadequate sensitivity (limits of detections between 0.3 and 5 μg/L) to quantify HAAs at the level usually found in drinking waters. Alternative headspace LPME methods for HAAs require heating (45 °C, 25 min) to derivatise and volatilise the esters but, by using solvent bar microextraction (SBME), the extraction/methylation takes place at room temperature without degradation of HAAs to trihalomethanes. Adequate precision (relative standard deviation of approximately 8%), linearity (0.1–500 μg/L) and sensitivity (10 times higher than the HS-SPME alternative) indicate that the SBME method can be a candidate for routine determination of HAAs in tap water. Finally, the SBME method was applied for the analysis of HAAs in tap and swimming pool water and the results were compared with those of a previous validated headspace gas chromatography–mass spectrometry method.      

Investigation of glow-discharge-induced morphology modifications on silicon wafers and chromium conversion coatings by AFM and rugosimetry

The effect of radiofrequency glow-discharge sputtering on the sample surface in terms of modifications in the surface morphology were investigated in this work by using atomic force microscopy (AFM) and rugosimetry measurements. The influence of GD operating parameters (e.g. rf power, discharge pressure and sputtering time) on surface roughening was investigated using two different types of samples: mirror-polished and homogeneous silicon wafers and chromate conversion coatings (CCCs). Surface morphology changes produced by GD sputtering into the sample surface were carefully investigated by AFM and rugosimetry, both at the original sample surface and at the bottom of GD craters using different GD experimental conditions, such as the sputtering time (from 1 s to 20 min), rf forward power (20–60 W for the Si wafer and 10–60 W for the CCC), and discharge pressure (400–1,000 Pa for the Si wafer and 500–1000 Pa for the CCC). In the present study, GD-induced morphology modifications were observed after rf-GD-OES analysis, both for the silicon wafers and the CCC. Additionally, the changes observed in surface roughness after GD sputtering were found to be sample-dependent, changing the proportion, shape and roughness of the micro-sized patterns and holes with the sample matrix and the GD conditions.     

HPLC Analysis and Pharmacokinetics of Picroside I in Dog Plasma

A sensitive and selective HPLC–UV method established for determination of picroside I in dog plasma has been used to study the pharmacokinetics of the drug after intravenous administration of three different doses. Sample pretreatment consists in deproteination by addition of acetonitrile; l-ascorbic acid was used to improve the stability of picroside I. The lower limit of quantification of picroside I was 0.05 μg mL⁻¹. The recovery of the method was up to 90%. After intravenous administration to dogs picroside I was mainly distributed in the central compartment and was rapidly eliminated from the plasma; the mean elimination half-life was 30.54 ± 4.34, 30.20 ± 3.78, and 34.02 ± 1.88 min for doses of 2.5, 5, and 15 mg kg⁻¹, respectively, and the respective values of AUC _0–∞ were 81.04 ± 19.95, 198.50 ± 27.77, and 586.44 ± 103.08 μg min mL⁻¹. The different doses had no significant effect on the main pharmacokinetic data and the kinetics seemed to be linear in dosage range 2.5–15 mg kg⁻¹.     

Application of hydrocyanic acid vapor generation via focused microwave radiation to the preparation of industrial effluent samples prior to free and total cyanide determinations by spectrophotometric flow injection analysis

A sample preparation procedure for the quantitative determination of free and total cyanides in industrial effluents has been developed that involves hydrocyanic acid vapor generation via focused microwave radiation. Hydrocyanic acid vapor was generated from free cyanides using only 5 min of irradiation time (90 W power) and a purge time of 5 min. The HCN generated was absorbed into an accepting NaOH solution using very simple glassware apparatus that was appropriate for the microwave oven cavity. After that, the cyanide concentration was determined within 90 s using a well-known spectrophotometric flow injection analysis system. Total cyanide analysis required 15 min irradiation time (90 W power), as well as chemical conditions such as the presence of EDTA–acetate buffer solution or ascorbic acid, depending on the effluent to be analyzed (petroleum refinery or electroplating effluents, respectively). The detection limit was 0.018 mg CN l⁻¹ (quantification limit of 0.05 mg CN l⁻¹), and the measured RSD was better than 8% for ten independent analyses of effluent samples (1.4 mg l⁻¹ cyanide). The accuracy of the procedure was assessed via analyte spiking (with free and complex cyanides) and by performing an independent sample analysis based on the standard methodology recommended by the APHA for comparison. The sample preparation procedure takes only 10 min for free and 20 min for total cyanide, making this procedure much faster than traditional methodologies (conventional heating and distillation), which are time-consuming (they require at least 1 h). Samples from oil (sour and stripping tower bottom waters) and electroplating effluents were analyzed successfully.     

Headspace gas chromatography–mass spectrometry for rapid determination of halonitromethanes in tap and swimming pool water

Halonitromethanes (HNMs) are one of the most cytotoxic and genotoxic classes found among the unregulated disinfection by-products formed by the reaction of chemical disinfectants with natural organic matter in water. Typical methods used to determine these compounds in water (mainly trichloronitromethane) are based on the Environmental Protection Agency (EPA) method 551.1 using liquid–liquid extraction. A fast and straightforward method for the determination of the nine HNMs in water has been developed using a static headspace (HS) coupled with gas chromatography–mass spectrometry (GC-MS). Important parameters controlling headspace extraction were optimised to obtain the highest sensitivity: 250 μL of methyl tert-butyl ether (as a chemical modifier) and 6 g of anhydrous sodium sulphate were added to the water sample; an oven temperature of 80 °C and an equilibration time of 20 min were also selected. The addition of a chemical modifier favoured the volatilisation of all HNMs, increasing their signals up to approximately four times. Under optimum conditions, the method developed provides limits of detection between 0.03 and 0.60 μg/L and a relative standard deviation of ∼6.0%. The developed method was validated and then compared with the reference method EPA 551.1 for the analysis of tap and swimming pool water. A good agreement in the results was observed, which corroborated the good performance of the proposed HS-GC-MS method.     

Analysis of erythritol in foods by polyclonal antibody-based indirect competitive ELISA

Sugar alcohols are widely used as food additives and drug excipients. Erythritol (INS 968) is an important four-carbon sugar alcohol in the food industry. Erythritol occurs naturally in certain fruits, vegetables, and fermented foods. Currently, HPLC and GC methods are in use for the quantification of erythritol in natural/processed foods. However, an immunoassay for erythritol has not been developed so far. We have utilized affinity-purified erythritol-specific antibodies generated earlier [9] to develop an indirect competitive ELISA. With erythritol–BSA conjugate (54 mol/mol; 100 ng/well) as the coating antigen, a calibration curve was prepared using known amounts of standard meso-erythritol (0.1–100,000 ng) in the immunoassay. Watermelon (Citrullus lanatus) and red wine were selected as the food sources containing meso-erythritol. The amount of meso-erythritol was calculated as 2.36 mg/100 g fresh weight of watermelon and 206.7 mg/L of red wine. The results obtained from the immunoassay are in close agreement with the reported values analyzed by HPLC and GC (22–24 mg/kg in watermelon and 130–300 mg/L in red wine). The recovery analyses showed that added amounts of meso-erythritol were recovered fairly accurately with recoveries of 86–105% (watermelon) and 85–93.3% (red wine). The method described here for erythritol is the first report of an immunoassay for a sugar alcohol. Figure Indirect competitive ELISA for quantitation of erythritol     

Phytochemical Study on Some Polyphenols of <Emphasis Type="Italic">Geranium pyrenaicum</Emphasis>

In order to continue our previous studies concerning Geranium pyrenaicum Burm. (Geraniaceae), we have performed spectrophotometric determinations and a HPLC study of some polyphenols. We have analyzed the dried Geranii pyrenaici herba (harvested from Cluj-Napoca, district of Cluj, Romania). We have established the content in flavonoids (0.316%), phenolic acids (0.099%), tannins (5.295%), and anthocyanins (12.030 mg/100 g vegetal product). We have identified and measured by HPLC the following compounds: hyperoside (21.61 μg/100 mg), ellagic acid (1810.44 μg/100 mg), isoquercitrine (11.197 μg/100 mg), and caftaric acid (76.83 μg/100 mg). We have also analyzed by HPLC a hydrolyzed sample of the same drug in which we have identified and measured: ellagic acid (4139.33 μg/100 mg), quercetol (29.65 μg/100 mg), kaempherol (41.48 μg/100 mg), and caftaric acid (20.721 μg/100 mg). __________ Published in Khimiya Prirodnykh Soedinenii, No. 4, pp. 322–324, July–August, 2005.     

Simultaneous stepwise injection-photometric determination of nitrite and nitrate ions in aqueous media

A procedure for the stepwise injection-photometric determination of nitrite and nitrate ions has been developed. The procedure employs their subsequent determination by the reaction of colored azo compound formation after the reduction of nitrate ions into nitrite ions on a cadmium reducer. The analytical ranges for nitrite and nitrate ions are 2–15 and 5–50 mg/L (sample volume 2 mL, analysis time 14 min).     

Exploiting the versatility of vacuum‐assisted headspace solid‐phase microextraction in combination with the selectivity of ionic liquid‐based GC stationary phases to discriminate Boswellia spp. resins through their volatile and semivolatile fractions

The frankincense resins, secreted from Boswellia species, are an uncommon example of a natural raw material where every class of terpenoids is present in similar proportions. Diterpenoids (serratol, incensole, and incensole acetate) are used to discriminate samples from different species and origins. Headspace solid‐phase microextraction has been used for frankincense analysis, although it requires long sampling time for medium‐ to low‐volatility markers; headspace solid‐phase microextraction under vacuum can overcome this limit. Gas chromatography is used for analysis but the separation of incensole and serratol needs polar stationary phases. In this study, we develop a method to discriminate frankincenses based on vacuum‐assisted headspace solid‐phase microextraction combined with fast gas chromatography‐mass spectrometry with ionic liquid–based stationary phases. The optimized conditions for solid samples were: air evacuation below 0°C, 15 min of incubation time, and 15 min of extraction time. Losses of volatiles due to vial air‐evacuation in the presence of the sample were minimized by sample amount above 100 mg and low sample temperature. Fast gas chromatography provides the baseline separation of all markers in 20 min. By applying vacuum sampling and fast gas chromatography, the total analysis was reduced to 50 min compared to 120 min (60 min sampling plus 60 min analysis) as previously reported. The method was successfully applied to commercial frankincense samples.     

Biodegradability of sol–gel silica microparticles for drug delivery

The biodegradability of porous sol–gel silica microparticles in physiological buffers has been investigated using a USP4 flow-through dissolution tester. In the open configuration, which most closely models in-vivo conditions, the particles dissolved rapidly at pH 7.4, with a rate dependent on the surface area and media flow rate. In the closed configuration, the fastest dissolving 4 mg silica sample was almost completely dissolved in 100 mL of buffer after 36 h. The initial dissolution rates appeared relatively linear but dropped off as dissolved SiO₂ concentrations approached 20–25 ppm. Addition of serum proteins acted to slow dissolution by 20–30%, suggesting a slower degradation in vivo. Silica microparticles administered for controlled release drug delivery would therefore be expected to be eliminated relatively rapidly from the body, depending on the sample size and local fluid flow conditions.