Development and Validation of a Gas-Chromatography–Mass-Spectrometry Method for Quantitative Determination of Ethylene Glycol and Diethylene Glycol in Solutions for Internal Use and Cough Syrups

Ethylene glycol (EG) is a cheap available solvent used mainly as antifreeze and in resin industry while diethylene glycol (DEG) is usually used in natural gas industry. In the past few years there was increasing reports about EG and DEG intoxication especially in Indonesia, Uzbekistan, Panama, India, Gambia and some African countries. The intoxication was due to using some pediatric cough syrups that are contaminated with these harmful solvents which lead to acute renal failure and death in some cases. World Health Organization declares that any limit of EG or DEG more than 0.1% considered a safety issue in the pharmaceutical products. In 2023, random pharmaceutical samples from Iraqi local market were tested by independent US laboratory (Valisure LLC) and there was an EG contaminated cough syrup with approximately 21 times more than the accepted limits. The above report encourage the conduction of this research to investigate the presences and the amount of EG and DEG in cold and cough syrups available in the Iraqi markets. In this study, 12 samples of cold and cough pediatric syrups of different origin were collected from the local market in Iraq and the levels of EG and DEG were measured by using gas chromatography with flame ionization detection (GC-FID) to estimate the presence and concentration of these compounds in the selected drugs. The results showed that only one sample was containing 0.65% DEG , while other samples were containing less than 0.1% or undetectable levels of either EG or DEG.

The contamination of cough syrups manufactured in Haryana was recently reported. The four cough and cold syrups so contaminated as to be banned by the WHO are Promethazine Oral Solution, Kofexmalin Baby Cough Syrup, Makoff Baby Cough Syrup, and Magrip N Cold Syrup. India-made cough and cold syrups are "potentially linked with acute kidney injuries and 70 children`s deaths" and will be distributed in The Gambia in 2022. According to the WHO, diethylene glycol and ethylene glycol are toxic to humans when consumed and can be fatal. This incident includes the prevalent proportion of cases with neurotoxic signs and symptoms

Recently there have been reports of the contamination of cough syrups with Diethylene Glycol (DEG). The consumption of such cough syrups has devastating effects on the health. In this paper we report evidence that Fourier transform infrared (FT-IR) and near-infrared (NIR) spectroscopic techniques are viable, simple, cost effective, rapid and fool proof methods for the identification and quantification of DEG in glycerin based cough syrups. The FT-IR and NIR spectra of the glycerin based cough syrup and up to 50:50 mixtures of DEG in cough syrup are recorded. The major peaks in the FT-IR spectrum of the cough syrup are assigned to the OH stretching (∼3300 cm−1), CH stretching (∼2900 cm−1), CH bending (1500–1200 cm−1) and C–O stretching (1200–900 cm−1) vibrational modes. In the FT-IR spectra of the mixtures, DEG contribute distinct peaks due to the vibrations of the C–O (920 cm−1) and OC2H4(892 cm−1) moieties of its backbone and form the basis of the DEG detection and quantification. The prominent peaks of the NIR spectra of cough syrup and DEG are assigned to the first overtones of OH and CH, and to the combination of OH and CH fundamental vibrations. Both FT-IR and NIR Partial Least Square (PLS) calibrations produced correlation coefficients of 0.98.

Recently there have been reports of the contamination of cough syrups with Diethylene Glycol (DEG). The consump- tion of such cough syrups has devastating effects on the health. In this paper we report evidence that Fourier transform infrared (FT-IR) and near-infrared (NIR) spectroscopic techniques are viable, simple, cost effective, rapid and fool proof methods for the identification and quantification of DEG in glycerin based cough syrups. The FT-IR and NIR spectra of the glycerin based cough syrup and up to 50:50 mixtures of DEG in cough syrup are recorded. The major peaks in the FT-IR spectrum of the cough syrup are assigned to the OH stretching (∼3300 cm −1 ), CH stretching (∼2900 cm −1 ), CH bending (1500-1200 cm −1 ) and C-O stretching (1200-900 cm −1 ) vibrational modes. In the FT-IR spectra of the mixtures, DEG contribute distinct peaks due to the vibrations of the C-O (920 cm −1 )a nd OC 2H4 (892 cm −1 ) moieties of its backbone and form the basis of the DEG detection and quantification. The prominent peaks of the NIR spectra of cough syrup and DEG are assigned to the first over- tones of OH and CH, and to the combination of OH and CH fundamental vibrations. Both FT-IR and NIR Partial Least Square (PLS) calibrations produced correlation coefficients of 0.98.

There has been a spate of drug related illnesses and deaths reported from Pakistan in the press this year. In November, at least 17 people died in the eastern city of Lahore after drinking toxic cough syrup [1]. Earlier in January, an adulterated heart medicine dispensed by the Punjab Institute of Cardiology claimed lives of at least 69 heart patients [2]. It seems appropriate to put these happenings in some perspective. Cough syrups have been at the center of several cases of mass poisonings around the world in recent years, often involving accidental or intentional use of industrial solvent - diethylene glycol - as a cheap sweetener by the manufacturers. Diethylene glycol is known for causing severe toxicity and death. The effects of such poisoning can be widened because cough syrup is frequently abused by people who use it to get high. It was the most likely scenario in Panama when at least 94 people died from taking cough syrups in 2006 [3]. A 7-year-old boy died in Manila in February this year after he suffered a seizure believed to have been caused by taking cough medicine [4]. Another five years old fell victim in Colorado, US on February 12 this year after taking the over-the-counter cough medicine to treat flu-like symptoms [5]. Similar incidents are reported from around the world totaling a massive number. Besides such clear incidences of toxicity, some seemingly innocuous medicines like iron supplements may account for more than 30% of reported pediatric medicine related fatalities. Antidepressants, cardiovascular medications, and methyl salicylate follow in frequency of pediatric pharmaceutical related deaths [6]. Let us take a few steps back and look at the bigger picture. Prescription drugs constitute the major treatment modality of scientific medicine today. Their utility has from the beginning, however, been overrated as manufacturers and promoters of these chemical drugs promised much more than they delivered. Far beyond not working, the drugs are known to cause incalculable side effects. Even when properly prescribed, these drugs have side effects that can be fatal. The situation would seem even worse considering the potential of human error. Lazarou [7] has shown that there might be as many as 106,000 deaths annually due to medicinal reactions happening in United States causing around $12 billion in economic losses. These are to be added to an annual figures of 98,000 deaths and &2 billion losses shown by Suh [8] as a result of human errors. But these put together would seem conservative in comparison to another study by Leape [9] has shown much higher estimates figured at 420,000 deaths and $200 billion in economic losses in a year. In a New England Journal of Medicine study, an alarming one in four patients have been shown to suffer observable side effects from the more than 3.34 billion prescription drugs filled in USA in 2002.[10] Besides the incalculable harm caused by the medicines as listed above, the death and suffering caused additionally due to hospital acquired infections, unnecessary procedures including surgeries, negligence and error brought forth by the healer reaches mind boggling dimensions. It would appear that causing harm is a rule rather than exception in the era of modern medicine. A look at the broader picture shows that medicine - as conventionally practiced – has failed to meet effectively many of the challenges of modern health care and instead of being part of a solution has become part of the problem.

A Selective Gas Chromatography–Tandem Mass Spectrometry Method for Quantitation of Ethylene and Diethylene Glycol in Paediatric Syrups

While Polyethylene glycol 3350 (PEG 3350) is approved by the USFDA for short term use by adults, it is commonly recommended for use in constipated children. Multiple reports of adverse events in children taking PEG 3350 raised safety concerns suggesting that low molecular weight species of PEG 3350 might be absorbed from the gut and cause side effects such as ethylene glycol (EG), diethylene glycol (DEG), and triethylene glycol (TEG). This article documents the development, validation, and application of analytical methods using GC-MS and GC-MS/MS for the quantitation of EG, DEG, and TEG in human plasma, serum, and urine. The analytical range for EG, DEG, and TEG was 2-20 µg/mL. The sample preparation process involves derivatization using N,O-bis(trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane in each biological matrices. Deuterated internal standards for each of the analytes were included to provide accurate quantitation of the glycol analytes. The validated methods were applied to analyze samples a pilot study of children taking PEG 3350. DEG and TEG were detected at levels below the limit of quantitation. In summary, a platform of analytical methods was developed to evaluate glycol analogs in urine, serum, and plasma clinical samples.

Identification and determination of diethylene glycol (DEG) in glycerin-based products was successfully achieved using FT-IR spectroscopy. Studied samples included 0.5% to 20% by mass DEGspiked into cough syrup, two paracetamol syrup formulations, and two food supplements. The characteristic DEGwavenumbers at 881 cm-1 and 1083 cm-1 were used for its quantitative determination in the studied samples. A very good accuracy in determining the DEG fraction was achieved with a mean error% of ±2.02% to ±7.69% upon using the corrected absorbance at 881 cm-1. The corrected absorbance at 1083 cm-1 band was used in the case of paracetamol formulations and resulted in a mean error% ranging from ±2.50% to ±10.28%. The values of limit of detection of the current method ranged from 0.051% to 0.068% DEG for all studied samples.

Dear Editor, Diethylene glycol (DEG) can be found in commercial products such as antifreeze, brake fluid, and lubricants. In addition, DEG has been found as a contaminant of raw materials in the production of pharmaceuticals. At least ten mass DEG poisoning events have occurred over the past 70 years. The first and largest outbreak, which resulted in 105 deaths, occurred in the United States in 1937 [1]. In 1967, a mass poisoning occurred in South Africa in which 7 children died [2]. In 1992 in Argentina, 29 people died after consuming propolis syrups that contained high DEG concentrations; the drug was widely commercialized in Argentina to treat mild upper respiratory tract infections [3]. Thereafter, pediatric medicinal syrups contaminated with DEG caused the deaths of 33 children in India in 1998 [4], and 85 children in Haiti in 1995–1996 [5]. The most recent outbreak took place in Panama in 2006, in which more than 100 people died due to DEG poisoning [1]. In spite of these repeated mass DEG poisonings, only a few analytical methods for DEG analysis, by gas chromatography (GC) [3] and GC–mass spectrometry (MS) [6–8], have been reported. There is a pressing need to establish and improve the methods for analysis of DEG, especially for postmortem human samples. As a result of our extensive experience in analysis of DEG in postmortem samples, we hereby report an important characteristic of DEG that we observed during sample extraction. Common chemicals, including ethylene glycol, DEG, fatty acids, phospholipids, and cholesterols, were of the highest chromatographic purity commercially available. GC was performed on a Shimadzu GC-14 A equipped with a Shimadzu CR 4A integrator (Shimadzu, Kyoto, Japan). A J&W DB-Wax column (30 m 9 0.53 mm i.d., 1.5 lm film thickness, Agilent, Santa Clara, CA, USA) was used. The injector temperature was set at 250 C and the flameionization detector (FID) was set at 250 C. An initial oven temperature of 110 C was held for 2 min before a temperature ramp of 8 C/min was used to reach the final temperature of 210 C. The carrier gas was nitrogen (12 cm/min). For thin-layer chromatography (TLC) analysis of lipid components, an Iatroscan TLC–FID apparatus (Mitsubishi Kagaku, Iatron, Tokyo, Japan) was used . Tissue samples (whole blood, liver, and kidney) were obtained from 15 victims of massive intoxication who ingested propolis syrups contaminated with DEG. Ten millilitres of blood was obtained by puncturing the femoral vein and was analyzed after 24 h. Fifty grams each of liver and kidney was taken from the victims and stored frozen at -20 C until analysis (3 days after collection). We isolated DEG from tissues by continuous Soxhlet extraction with methanol for 12 h. The extract obtained from each sample was purified using a charcoal column and evaporated to 1 ml. Then 10 ll of the concentrated extract was used for analysis of DEG by GC–FID. When the methanol extract was stored in a refrigerator at 2–4 C for several hours, a semi-crystalline substance appeared, adhering to the glass wall of the test tube. When subjected to GC analysis, the supernatant fraction of the This article is for the special issue TIAFT2012 edited by Osamu Suzuki.

The silent epidemic of substandard and falsified medicines in low- and middle-income countries: heed lessons from the tragic deaths of children in Indonesia

Introduction. Ethylene glycol and diethylene glycol are highly toxic compounds with adverse effects on the central nervous system and paren-chymatous organs. A number of excipients such as glycerol, propylene glycol and polyethylene glycol may be the source of these impurities in phar-maceutical preparations. It is therefore necessary to control the ethylene glycol and diethylene glycol content for the safe use of such medicinal prod-ucts. The regulatory documents provide such methods of their analysis as thin-layer chromatography, high-performance liquid chromatography and gas chromatography. These methods differ in sensitivity and selectivity, which requires an assessment of their applicability for the analysis of toxic impuri-ties of ethylene glycol and diethylene glycol in various dosage forms. Objective of the study. On the example of a sample of solution for subcutaneous and intravenous administration containing insulin aspart 100 U/mL, to conduct a comparative assessment of the applicability of high-performance liquid chromatography and gas chromatography methods in the control of the content of impurities of ethylene glycol and diethylene glycol. Material and Methods. The object of the study was a sample of solution for subcutaneous and intravenous administration containing insulin aspart 100 U/mL. This preparation contains glycerol as a source of the impurities sought. The tests were carried out on Agilent brand liquid chromatograph and Chrozen brand gas chromatograph. Results. Methods for quantitative determination of ethylene glycol and diethylene glycol by high-performance liquid chromatography and gas chroma-tography in a sample of solution for subcutaneous and intravenous administration were developed. According to the results of validation experiments the applicability of the developed analytical technique (gas chromatography method) for quantitative determination of ethylene glycol and diethylene glycol in the investigated preparation was confirmed. Conclusions. This study demonstrated the possibility of using both gas chromatography and high-performance liquid chromatography methods to control the content of ethylene glycol and diethylene glycol impurities in a sample of solution for subcutaneous and intravenous administration. Howev-er, in order to comply with the regulatory requireнапрments (not more than 0.1% per drug) presented in regulatory documents, it is more appropriate to use the gas chromatography method due to its higher sensitivity characteristics.

In September 2006, a Panamanian physician reported an unusual number of patients with unexplained acute renal failure frequently accompanied by severe neurological dysfunction. Twelve (57%) of 21 patients had died of the illness. This paper describes the investigation into the cause of the illness and the source of the outbreak. Case-control and laboratory investigations were implemented. Case patients (with acute renal failure of unknown etiology and serum creatinine > 2 mg/dl) were individually matched to hospitalized controls for age (+/- 5 years), sex and admission date (< 2 days before the case patient). Questionnaire and biological data were collected. The main outcome measure was the odds of ingesting prescription cough syrup in cases and controls. Forty-two case patients and 140 control patients participated. The median age of cases was 68 years (range: 25-91 years); 64% were male. After controlling for pre-existing hypertension and renal disease and the use of angiotensin-converting enzyme inhibitors, a significant association was found between ingestion of prescription cough syrup and illness onset (adjusted odds ratio: 31.0, 95% confidence interval: 6.93-138). Laboratory analyses confirmed the presence of diethylene glycol (DEG) in biological samples from case patients, 8% DEG contamination in cough syrup samples and 22% contamination in the glycerin used to prepare the cough syrup. The source of the outbreak was DEG-contaminated cough syrup. This investigation led to the recall of approximately 60 000 bottles of contaminated cough syrup, widespread screening of potentially exposed consumers and treatment of over 100 affected patients.

Ethylene glycol (EG) and diethylene glycol (DEG) are hazardous compounds if ingested by the human body. Testing conducted by the Indonesian National Agency of Drug and Food Control (BPM) identified contamination of EG and DEG in glycerin and propylene glycol, which are utilized as solubility enhancers in syrup-based pharmaceuticals. This study aims to analyze EG and DEG contamination in children’s syrup drug samples using a Gas Chromatography-Mass Spectrometry (GC-MS) instrument. This study presents an analytical method for detecting EG and DEG contamination. Based on BPOM regulations governing the control a determination of EG and DEG. Several syrup drug samples, which are listed in the BPOM drug withdrawal list under Number HM.01.1.2.11.22.240, were each weighed at 5 grams, transferred into a 50 mL volumetric flask, 30 mL of methanol was added, and the mixture was sonicated for 5 minutes. Subsequently, solvent was added up to the calibration mark. Five out of eight samples were found to contain EG and DEG at concentrations of less than 0.1%. Validation of the GCMS method for EG and DEG compounds yielded accuracy values with recovery rates between 98% and 101%. The acquisition test results produced a relative standard deviation (RSD) value of 0.87. The linearity test showed a correlation coefficient (r) of 0.955. The limit of detection (LOD) for EG was 0.26 ng/mg, and for DEG it was 0.51 ng/mg. The limit of quantification (LOQ) for EG was 0.86 ng/mg, and for DEG it was 1.69 ng/mg. This validation indicates that the analytical method exhibits high accuracy and sensitivity.

Due to a fatal accident that occurred in 2006 for diethylene glycol (DEG) adulterated in glycerin, the Pharmaceutical and Food Safety Bureau in the Ministry of Health, Labour and Welfare (MHLW) has notified the partial revision of the purity test of Glycerin in the Japanese Standards of Quasi-Drug Ingredients (JSQI) 2006 at the Notification No. 1221004 on February 21, 2008. Since the fatal accidents caused by the contamination of syrup with DEG also occurred in 2009, USP proposed the purity test of concentrated glycerin, glycerin and propylene glycol about ethylene glycol (EG) and DEG as the adulterant instead of the purity test of concentrated glycerin and glycerin about DEG on May 21, 2009 to the Pharmacopoeial Discussion Group. In response to this, the Expert Committee on Excipients on JP made the decision to establish the new modified method. This paper proposes the new modified method of DEG including EG for the purity test in concentrated glycerin, glycerin and propylene glycol in JSQI 2006. This analytical method was the gas chromatographic method by using the fusedsilica column 0.32 mm×30 m coated with 14% cyanopropylphenyl/86% methylsilicon polymer and the column temperature for injecting at a constant temperature of about 100oC and raising at the ratio of 7.5oC per minute to 220oC. The retention times of EG, propylene glycol, DEG and glycerin were 2.45, 2.78, 6.02 and 7.66 minutes, respectively. The working curves of EG and DEG were the good correlation between their concentrations of 2.5 to 80 μg/ml and the peak areas. The resolutions between EG and DEG, and between DEG and glycerin were not less than 70 and not less than 20, respectively. Also those between EG and propylene glycol, and between propylene glycol and DEG were not less than 6 and not less than 60, respectively. The quantitation limits of EG and DEG in glycerin were 0.005% and 0.01%.

Clinical, Laboratory, Diagnostic, and Histopathologic Features of Diethylene Glycol Poisoning—Panama, 2006