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 Table of Contents  
ORIGINAL ARTICLE
Year : 2015  |  Volume : 1  |  Issue : 1  |  Page : 48-53

Comparison of the Concentrations of Lidocaine in Different Body Fluids/Tissues after Subarachnoid Space and Intravenous Administration of a Lethal Dose of Lidocaine


School of Forensic Medicine, Shanxi Medical University, Taiyuan, China

Date of Web Publication29-May-2015

Correspondence Address:
Keming Yun
School of Forensic Medicine, Shanxi Medical University, 56 Xinjian South Street, Taiyuan 030001, Shanxi
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2349-5014.157907

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  Abstract 

The objective of the study was to compare the concentration of lidocaine in different body fluids/tissues after subarachnoid space and intravenous administrations of a lethal dose of lidocaine. Totally 18 dogs were used in the experiment. Six dogs were given subarachnoid anesthesia, another were given an intravenous injection of a dose of 75 mg/kg weight of lidocaine hydrochloride in 5 min and the last 6 dogs were used as the blank control dogs and given a subarachnoid space injection or a femoral artery injection of the same volume of sodium chloride. As soon as its vital signs disappeared, each dog was dissected and the specimen, such as brain, cerebrospinal fluid (CSF) in lateral ventricle, CSF in subarachnoid space, spinal cord (cervical spinal cord, thoracic spinal cord, lumbar spinal cord, and waist spinal cord), heart, lung, liver, spleen, kidney, bile, urine, heart blood, peripheral blood, muscle in injection location, and muscle in no injection location, were collected for analysis of lidocaine immediately. Analysis was performed with gas chromatography-mass spectrometry (GC-MS). From the maximum to the minimum, the order of lidocaine concentration detected in the subarachnoid space-administered dogs was as follows: CSF in subarachnoid space, waist spinal cord, thoracic spinal cord, CSF in lateral ventricle, lumbar spinal cord, cervical spinal cord, lung, kidney, muscle in injection location, heart, brain, spleen, heart blood, liver, peripheral blood, bile, muscle in no injection location, and urine. The order of lidocaine concentration detected in the intravenously administered dogs was as followed: Kidney, heart, lung, spleen, brain, liver, peripheral blood, bile, heart blood, cervical spinal cord, thoracic spinal cord, muscle in injection location, lumbar spinal cord, muscle in no injection location, CSF in subarachnoid space, urine, and CSF in lateral ventricle. The maximum concentration of lidocaine was detected in the subarachnoid space CSF of subarachnoid space-administered dead dogs, while in intravenously injected dead dogs, the maximum concentration of lidocaine was detected in the kidney. Our study provides some useful data for the forensic identification of epidural anesthesia accidents to decide the way the lidocaine enters the body.

Keywords: Anesthesia accident, distribution of lidocaine, forensic medicine, forensic toxicological analysis, intravenous administration, intravenous injection, lidocaine, postmortem distribution, subarachnoid administration, subarachnoid anesthesia


How to cite this article:
Zhang N, Sun J, Wei Z, He W, Jin G, Zhang X, Gao P, Wang L, Yun K. Comparison of the Concentrations of Lidocaine in Different Body Fluids/Tissues after Subarachnoid Space and Intravenous Administration of a Lethal Dose of Lidocaine. J Forensic Sci Med 2015;1:48-53

How to cite this URL:
Zhang N, Sun J, Wei Z, He W, Jin G, Zhang X, Gao P, Wang L, Yun K. Comparison of the Concentrations of Lidocaine in Different Body Fluids/Tissues after Subarachnoid Space and Intravenous Administration of a Lethal Dose of Lidocaine. J Forensic Sci Med [serial online] 2015 [cited 2019 May 22];1:48-53. Available from: http://www.jfsmonline.com/text.asp?2015/1/1/48/157907


  Introduction Top


Lidocaine is used widely as a local anesthetic and antiarrhythmic drug. [1] In a human clinical setting, a dose of no more than 11-15 mg/kg in a single injection would be used for epidural anesthesia. [2] Because of the special structure of spinal cord space and some technical factors, some epidural anesthesia accidents occurred, which were due to an accidental subarachnoid space or intravenous administration of lidocaine for intended epidural anesthesia. [3],[4],[5],[6],[7],[8],[9] The postmortem distribution of lidocaine in victims is more useful for the forensic identification of epidural anesthesia accidents, but there have been a few reports on the postmortem distribution of lidocaine in humans who died of epidural anesthesia accidents, [10],[11],[12] and the postmortem distribution or the concentration of lidocaine in different body fluids/tissues after subarachnoid space and intravenous administrations of a lethal dose of lidocaine are also unclear. [13] Keming Yun and Jiong Yan [14],[15] had established a model and investigated the distribution of lidocaine by a  thin-layer chromatography scanning (TLCS) in dogs after a subarachnoid administration to judge accidents in epidural anesthesia, but the specificity and precision of lidocaine determination by the TLCS analysis was not very good, compared to the gas chromatography (GC), GC-mass spectrometry (GC-MS), or high-performance liquid chromatography-MS (HPLC-MS) determination of lidocaine. [16],[17],[18] In this study, we use the dog animal models to compare the concentrations of lidocaine in different body fluids/tissues after subarachnoid space and intravenous administrations of a lethal dose of lidocaine, provide some useful data for the identification of epidural anesthesia accidents to decide the way lidocaine enters the body.


  Materials and Methods Top


Reagents

Lidocaine standard substance (10342-0001) and internal standard SKF 525A were purchased from the  National Institute for The Control of Pharmaceutical and Biological Products, Beijing, People's Republic of China. Lidocaine hydrochloride injection (0.4 g/20 mL) was obtained from Fosun Zhaohui Corporation (Shanghai, People's Republic of China). Lidocaine and internal standard SKF 525A solutions all were prepared at a concentration of  1 ΅g/΅L, respectively.

Experiment

Animals


Adult male dogs (10-14 months) weighing 15-20 kg were obtained from the laboratory animal center of Shanxi Medical University. Animals were individually housed followed the guidelines of the animal committee. Before experiments, dogs were fasted overnight. The protocol was permitted by the Ethical Review Committee, Shanxi Medical University, Shanxi, People's Republic of China.

Animal models

Establishment of a fatal model of subarachnoid space administration of lidocaine

Nine dogs were randomly allocated to positive samples (n = 6 per group) and negative control groups (n = 3 per group). All the dogs were positioned laterally on the table throughout the experiment, and the BL-biological function experimental system (Taimeng, Chengdu, People's Republic of China) was used to monitor the electrocardiogram, breathing changes and blood pressure following its protocol. According to the method reported by Keming Yun, [14] the dog body was compressed to abdomen side. Then a spinal needle was put through the intervertebral space L 2 ~ 3 , the mater of spinal cord, and the spinal arachnoid matter into subarachnoid space. A small amount of clear spinal fluid was aspirated to make sure the needle had penetrated the subarachnoid space. 75 mg/kg of lidocaine hydrochloride was injected into the subarachnoid space (L 2 ~ 3 ) in the positive groups in 5 min, while the same volume of sodium chloride was injected into the subarachnoid space (L 2 ~ 3 ) in the negative control groups. After the subarachnoid space injection of lidocaine had been given, the dogs were monitored continuously until death.

Establishment of a fatal model of intravenous administration of lidocaine

As in the above method, the six dogs in the positive groups were given a femoral artery injection of 75 mg/kg of lidocaine hydrochloride and the three dogs in the negative control groups were given a femoral artery injection of the same volume of sodium chloride.

Collection of samples

In the two positive groups, as soon as the vital signs (heart rate, respiration, blood pressure) disappeared, the dogs were anatomized, and specimens-the brain, cerebrospinal fluid (CSF) in lateral ventricle, CSF in subarachnoid space, spinal cord (cervical spinal cord, thoracic spinal cord, lumbar spinal cord, and sacral spinal cord), heart, lung, liver, spleen, kidney, bile, urine, heart blood, peripheral blood, muscle in the injection location, and muscle 20 cm from the injection location-were collected for the determination of lidocaine, immediately by GC-MS. Negative control group dogs were monitored and given a gas embolism by femoral artery injection of gas, and specim ens were collected as blank samples.specimens were collected from the two model dogs mentioned above (gas to death of which tissue and body fluid were collected as the blank sample).

Sample analysis

GC-MS chromatography

A GC-MS chromatography instrument (Thermo Finnigan model trace DSQ-MASS Spectrometer ) was used to determine lidocaine in the tissues and fluids of the dogs. [16] The GC-MS conditions were as following: The column was quartz (DB-5 capillary column, 30 m Χ 0.25 mm Χ 0.25 μm); helium flow was 1.5 mL/min; the initial oven temperature was 150°C, maintained at 1 min, then increased to 280°C. At 10°C/min, the final temperature was 280°C and held for 2 min.(280°C at the speed of 10°C/min, the final temperature was 280°C and maintained for 2min).

Extraction of lidocaine in tissues/body fluids

One gram (1 g) of tissue and 1 mL of distilled water were homogenized intensively and the matter was transferred to a glass tube. 1 mL of body fluid and 1 mL of distilled water were added to a glass tube. Then 12 μg of an internal standard solution and 2 mL of 1% HCL were added to the glass tube. After mixing for 15 min, the tube was centrifuged at 3000 rpm for 15 min and the supernatant acid was transferred to a glass tube. Then 2 mL of 0.5 M NaOH and 5 mL of ethyl ether were added and the ethyl ether layer was evaporated to dryness under nitrogen pressure. The residue was dissolved in 50 μL of alcohol of which 1 μL of the solution was injected into the GC-MS.

Qualitative and quantitative analysis of lidocaine

Lidocaine was analyzed qualitatively and quantitatively with the GC-MS. [16] Recorded by a GC-MS-selected ion monitoring (SIM) model, lidocaine in tissues and in body fluids was quantitatively analyzed by an internal standard and calibration curve. The working curves of lidocaine in different tissues and body fluids were established as described, and the linear equation, linear range R 2 , and the limit of quantification (LOQ) were obtained. The extraction recovery of lidocaine was done by the quantitative analysis of lidocaine added to blank blood, brain, CSF, and spinal cord. The concentrations of lidocaine in all collected tissues and body fluids were calculated, and the ratios of specimen lidocaine concentration to peripheral blood lidocaine concentration were also investigated.

Data analysis

All data were expressed as mean ± (standard deviation) SD. Statistical evaluation was performed with  SPSS Statistics Version 15 (SPSS statistics version 15 for windows). One-way analysis of variance (ANOVA) followed by the Student-Newman-Keuls test (SNK) were used to compare the differences between groups.


  Results Top


GC-MS analysis

The GC-MS chromatogram for dog blood is shown in [Figure 1]. The working curves for quantitative analysis of lidocaine in different tissues or body fluids are showed in [Table 1] and [Table 2]. The linear range was 2-200 μg/mL. The LOQ was 0.05-0.08 μg/mL in the tissues and body fluids, respectively.
Table 1: Working curves of lidocaine in different tissues/ fluids


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Table 2: Recovery of lidocaine in blood, brain, CSF, and spinal cord (n=3, x −±SD, %)


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Figure 1: GC-MS chromatogram of lidocaine and SKF525A (a) total ion current (TIC) GC-MS chromatogram (b) SIM GC-MS chromatogram (c) Full-scan mass spectra of lidocaine. Lidocaine tr: 8.08 min; internal standard SKF525A tr; 12.12 min.

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Animal model

For 1 ~ 2 min after the intravenous administration of lidocaine, the positive dogs showed contracted pupils, respiratory arrest, aphonia, muscle tremble, opisthotonos, incontinence of urine and excrement, and  extensive sensory block, and their  electrocardiac signals, blood pressure, and respiration disappeared on average in 8 min, 6.5 min, and 6.5 min, respectively. For 1 ~ 2 min after the subarachnoid space administration of lidocaine, the positive dogs showed opisthotonos, incontinence of urine and excrement, and their electrocardiac signals, blood pressure, and respiration disappeared on average in 14 min, 12 min, and 17 min respectively. All the dogs in the two negative control groups showed good vital signs. None of them died before the final gas embolism.

Comparison of lidocaine concentrations in in body fluids/tissues between different models

The concentrations of lidocaine in different tissues and body fluids after subarachnoid space and intravenous administrations of a lethal dose of 75 mg/kg lidocaine are shown in [Table 3] and [Table 4] and [Figure 2] and [Figure 3]. The ratios of tissue or body fluid lidocaine concentration to peripheral blood lidocaine concentration are also shown in [Table 3] and [Table 4].
Table 3: Lidocaine concentration in fatal dogs after a subarachnoid administration of 75 mg/kg lidocaine


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Table 4: Lidocaine concentration in fatal dogs after an intravenous administration of 75 mg/kg lidocaine


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Figure 2: Lidocaine concentration in fatal dogs after a subarachnoid administration of 75 mg/kg lidocaine. Tissues and body fluids: 1. Brain; 2. cervical spinal cord; 3. thoracic spinal cord; 4. lumbar spinal cord; 5. sacral spinal cord; 6. dorsal CSF; 7. lateral ventricle CSF; 8. heart blood; 9. peripheral blood; 10. heart; 11. lung; 12. liver; 13. spleen; 14. kidney; 15. muscle in injection location; 16. muscle 20 cm from injection location; 17. urine; 18. bile

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Figure 3: Lidocaine concentration in fatal dogs after an intravenous administration of 75 mg/kg lidocaine. Tissues and body fluids: 1. Brain; 2. cervical spinal cord; 3. thoracic spinal cord; 4. lumbar spinal cord; 5. sacral spinal cord; 6. dorsal CSF; 7. lateral ventricle CSF; 8. heart blood; 9. peripheral blood; 10. Heart; 11. Lung; 12. liver; 13. spleen; 14. kidney; 15. muscle in injection location; 16. muscle 20 cm from injection location; 17. urine; 18. bile

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  Discussion Top


Animal model

As a local anesthetic and antiarrhythmic drug, no more than 11-15 mg/kg of lidocaine would be used in a single injection in a human clinical setting. [2] As reported by Yan Jiong, [15] in our previous experiment, dogs were given a subarachnoid space injection of 12.67 mg/kg lidocaine to establish a fatal model of subarachnoid space administration of lidocaine, in which the blood pressure, respiration, and electrocardiac signals of the dogs disappeared on average within 23.8 min, 16.4 min, and 18.6 min, respectively, after the injection, and the concentrations detected follow the order: Spinal subarachnoid space CSF, lateral ventricle CSF, the upper breast spinal cord, waist spinal cord, injection location spinal cord,   breast spinal cord, cervical spinal cord, medulla oblongata, injection location muscle, brain, blood, bile, urine, lung, liver, kidney, hearts, spleen, and non-injection location muscle [Figure 4]. In the present study, we tried to establish the fatal model of intravenous administration of lidocaine. During the preliminary experiment, it was found that no less than 75 mg/kg lidocaine in a single intravenous injection could account for dog death. Thus the same dose of lidocaine, 75 mg/kg, was chosen to establish the fatal model of intravenous administration and the fatal model of subarachnoid space administration for dogs. The present study showed that the tendency of lidocaine distribution and vital sign changes in 75 mg/kg subarachnoid space-administered dogs was similar to that in the 12.34 mg/kg dose-administered dogs. Therefore, the present models can be used to compare the concentrations of lidocaine in different body fluids/tissues after subarachnoid space and intravenous administrations.
Figure 4: Lidocaine concentration in fatal dogs after a subarachnoid administration of 12.67 mg/kg lidocaine.[15] Tissues and body fluids: 1. Brain; 2. cervical spinal cord; 3. thoracic spinal cord; 4. lumbar spinal cord; 5. sacral spinal cord; 6. dorsal CSF; 7. Lateral ventricle CSF; 8. heart blood; 9. heart; 10. lung; 11. liver; 12. spleen; 13. kidney; 14. muscle in injection location; 15. muscle 20 cm from injection location; 16. urine; 17. bile.

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Comparison of the concentrations of lidocaine in different body fluids/tissues after subarachnoid space and intravenous administrations

Because epidural anesthesia accidents occur due to an accidental subarachnoid space or intravenous administration of lidocaine at the time of intended epidural anesthesia, [3],[4],[5],[6],[7],[8],[9] it is very important in the process of forensic identification to decide the way the lidocaine enter the body.

Compared to each other, the postmortem distribution or body fluid/tissue concentration of lidocaine showed different tendencies. From the maximum to the minimum, the order of lidocaine concentrations detected in subarachnoid space-administered dogs was CSF in subarachnoid space, waist spinal cord, thoracic spinal cord, CSF in lateral ventricle, lumbar spinal cord, cervical spinal cord, lung, kidney, muscle in injection location, heart, brain, spleen, heart blood, liver, peripheral blood, bile, muscle in no injection location, and urine; the order of lidocaine concentrations detected in intravenous administered-dogs was kidney, heart, lung, spleen, brain, liver, peripheral blood, bile, heart blood, cervical spinal cord, thoracic spinal cord, muscle in injection location, lumbar spinal cord, muscle in no injection location, CSF in subarachnoid space, urine, and CSF in lateral ventricle. It was induced that the postmortem distribution or body fluid/tissue concentration of lidocaine in dogs may reflect the way lidocaine enters the body.

The maximum concentration was in the CSF and spinal cord after the subarachnoid administration, the body fluid to blood concentration ratios were from 52.4 ± 55.5 to100.8 ± 181.9, the minimum concentration was in urine, and the body fluid to blood concentration ratio was 0.2, which was similar to the Lu Yuanxu et al. [12] and Yan Jiong et al. [15] [Figure 4] reports, as the maximum concentration of the drug was in CSF and spinal fluid, but it was not the same with the study of Sakata et al., [19] as the concentrations of lidocaine in the brain were from five to ten times higher than those in the other tissues of individuals who died from anesthetic accidents. So maximum spinal and CSF lidocaine concentrations may be the evidence for lidocaine entering the body by the subarachnoid space.

The maximum concentration was in the kidney after intravenous administration, and the tissue-to-blood concentration ratio was  24.3 ± 26.1; the minimum concentration was in the CSF and spinal cord, and the body fluid or tissue-to-blood ratio was 0.2:1.1. Poklis [20] also observed that the maximum concentration of lidocaine after intravenous administration was in the kidney. The blood flow to the tissues is very different, and the transport of drug from blood to certain tissues is very rapid and much more, especially in the kidney, heart, lung, spleen, and brain. [21] The drug is distributed preferentially into tissues that have the highest affinity for it and high blood flow. [22] Thus, the maximum concentration being in the kidney may offer evidence for the way lidocaine enters the body on intravenous administration.


  Conclusion Top


In conclusion, the maximum lidocaine concentration induced during our study was in the spinal cord and CSF and the minimum lidocaine concentration in the urine after the subarachnoid space administration, while the maximum lidocaine concentration was in the kidney and minimum lidocaine concentration in the spinal cord and CSF after the intravenous administration. Our study provides some evidence for the forensic identification of epidural anesthesia accidents to judge the way lidocaine enters the body. However, there were a great variety of tissue and body fluid lidocaine concentrations in our study. The great variety in time from lidocaine administration to death may have contributed to a wide range of SD, which should be taken into account, and which needs further study.


  Acknowledgments Top


We thank the staff of the School of Forensic Medicine, Shanxi Medical University. This research was funded by the National Natural Science Foundation Council of China (No. 81172906), the National Key Technology R&D Program of China (No.2012BAK02B02-2) and International technology cooperation plan project in Shanxi Province (No.2012081053).

 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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