Pre-analytical considerations of condom traces: A review of composition, background, transfer and persistence.

Today, rape and sexual assault cases are mainly solved using evidence such as medical evidence or DNA analysis. Condom traces have been found to be present in 10% of assaulted women, when no DNA is found [1]. Numerous studies have emphasized the interest of analysing the composition of male condoms and their traces, and developing specific methods for the analysis of this type of evidence. However, transfer and persistence of condom traces in a specific matrix are rarely referenced. Therefore, forensic scientists have no complete knowledge of the trace and what could be expected in a real case. The purpose of this article is to review the literature addressing the composition of condoms and their traces as well as its influence on the transfer and persistence from a forensic point of view. Peer-reviewed literature, patents, professional literature, data from international administrations and international organisations' reports have been used to track the composition and the problematics of transfer and persistence of condom traces. The results of this review show that the composition of male condoms and their traces are complex systems, with numerous compounds originating from the condom at the moment of the transfer and evolving over time according to specific persistence patterns. Although numerous types of analyses have already been proposed and tested for condom traces, forensic evidence considerations have not been fully studied yet. Considering the fact that sexual assaults without the detection of DNA are increasingly frequent, there is a definite medical and forensic need to improve our knowledge of the processes involved in the development of condom traces in order to better understand analytical results.

Comparative analysis of condom lubricants on pre & post-coital vaginal swabs using AccuTOF-DART.

In this study we demonstrate the use of Direct Analysis in Real Time Mass Spectrometry (DART) as a powerful tool for detection of nonoxynol in vaginal fluid post contact with a condom, enabling rapid tracing and added evidences in sexual assault crimes. Vaginal fluid was sampled using cotton swabs and glass rods and measured directly with DART. Sample preparation using water, hexane, methanol, and dichloromethane extraction, was explored for comparison and optimization of signals. Nonoxynol was detected up to eight hours after sampling. Optimal sampling conditions and mass spectrometry parameters are reported and discussed.

Spermicidal Constituents of Ethanolic Extract of Sacoglottis gabonensis Stem Bark.

AIM: To isolate the spermicidal constituents of Sacoglottis gabonensis. MATERIALS AND METHODS: The ethanolic extract with partitioned fractions of Sacoglottis gabonensis stem bark were subjected to sperm immobilization assay. The most active EtOAc fraction was further purifi ed by column and Semi-Preparative High Performance Liquid Chromatography to give compounds which were characterized by spectroscopic methods (UV, LC/MS, and NMR). The compound(s) was also tested for sperm immobilization activity. RESULTS: The ethanolic extract showed 100% signifi cant (p < 0.05) sperm immobilization activity at a concentration of 30 mg/mL at 20 s compared to both negative and positive controls. The most active ethyl acetate fraction yielded methyl 3,5-dihydroxy-4-methoxybenzoate, eriodictyol and bergenin. Bergenin had 100% sperm immobilization activity at 20 mg/mL in 60 s which was signifi cant (p < 0.05) also when compared to the positive and negative control while methyl 3,5-dihydroxy- 4-methoxybenzoate, eriodictyol were not active. CONCLUSION: The active spermicidal constituent in Sacoglottis gabonensis stem bark extract is bergenin. However, methyl 3,5-dihydroxy-4-methoxybenzoate and eriodictyol showed no activity. This plant is known for its aphrodisiac action; hence, caution may have to be exercised in its use because of its spermicidal eff ect.

Analysis of sexual assault evidence: statistical classification of condoms by ambient mass spectrometry.

Desorption electrospray ionization mass spectrometry (DESI-MS) and easy ambient sonic-spray ionization mass spectrometry (EASI-MS) are employed here in the forensic analysis of chemical compounds found in condoms and relative traces, and their analytical performances are compared. Statistical analysis of data obtained from mass spectra only was applied in order to obtain classification rules for distinguishing ten types of condoms. In particular, two supervised chemometric techniques [linear discriminant analysis (LDA) and soft independent modeling of class analogy (SIMCA)] were carried out on absolute and relative intensity values to test the performances of statistical models in terms of predictive capacity. The achieved classification of samples was excellent because of the high prediction percentages of the method used both for DESI and EASI mass spectrometry analyses, confirming these two as potential ambient ionization techniques for forensic analyses in case of sexual assault crimes. EASI-MS showed 99% prediction ability for LDA using relative data and 100% prediction ability for SIMCA using both absolute and relative ones, while DESI showed 94% prediction ability for both LDA and SIMCA. The absence of any sample preparation technique gives advantages in terms of sample preservation and reduced contamination, allowing successive analyses to be performed on the same sample by other techniques.

Detection of the spermicide nonoxynol-9 via GC-MS.

The spermicide nonoxynol-9 is actually a complex mixture of dozens of closely related amphiphilic compounds, and the chemical properties of this assortment significantly hamper its characterization by GC-MS. The inability to perform routine GC-MS testing on nonoxynol-9 has limited its evidentiary value in forensic casework, which relies heavily on this technique for analysis. A disturbing trend in sexual assault is the use of condoms by assailants, to avoid leaving behind DNA evidence that can connect a perpetrator to a victim. This observation necessitates the development of alternative methods for the analysis of trace evidence that can show causal links between a victim and a suspect. Detection of lubricants associated with sexual assault is one such way to establish this connection. The development of GC-MS methods that permit facile detection of both nonoxynol-9 alone and nonoxynol-9 extracted from other complex matrices that have potential as trace evidence in sexual assault is reported. A detection limit of 2.14 µg of nonoxynol-9 is demonstrated, and a detailed mass spectral profile that elaborates on what is known of its structure is provided.

Contraceptive efficacy and safety of HerbOshield™ vaginal gel in rats.

BACKGROUND: Spermicides represent one of the methods of contraception. The synthetic agents available as spermicides produce severe side effects. Hence, there is a need to replace these agents with safe and effective agents such as plant-based contraceptive agents. STUDY DESIGN: The objective of the present study was to develop and evaluate a stable, safe, effective and easily acceptable contraceptive delivery system containing herbal drug. Efforts were made to evaluate the contraceptive potential of the hydroalcoholic extract from the seeds of Annona squamosa Linn. and the vaginal gel HerbOshield™ containing the extract. RESULTS: Spermicidal effect was evaluated in vitro using healthy human spermatozoa and in vivo in rats. The in vitro results demonstrated that HerbOshield™ vaginal gel is an effective spermicide. At a 100-mg/mL dose, complete immobilization of human spermatozoa was observed within 20 s. None of the treated animals conceived, indicating 100% contraceptive effect as compared to Gynol II, a nonoxynol-9-containing marketed formulation, which showed only 67% contraceptive effect in vivo. HerbOshield™ vaginal gel was found to be safe in animals during a 14-day toxicity study. CONCLUSIONS: HerbOshield™ vaginal gel was found to be safe and effective in rats and could be developed as a potential vaginal contraceptive for future use in humans.

Nuevas opciones en anticoncepción: posible uso espermicida de plantas colombianas / Colombian plants with spermicidal activity, new options in anticonception: brief review

En la actualidad existe una creciente preocupación por la salud sexual y reproductiva de los adolescentes, ya que pese a la existencia de diferentes métodos anticonceptivos el número de embarazos no deseados va en aumento. Entre los productos disponibles se encuentran los espermicidas, que son un método totalmente controlado por la mujer y con características ventajosas respecto a los anticonceptivos de uso continuo, pero que tiene como desventaja la provocación de irritaciones en el epitelio vaginal debido a un efecto tensoactivo sobre la membrana celular, lo que podría facilitar la transmisión de infecciones. En la búsqueda de nuevas alternativas, se ha encontrado que una amplia variedad de plantas tiene actividad espermicida, lo que permite pensar en una propuesta interesante para un anticonceptivo de origen vegetal, que constituya una herramienta clave en la prevención de embarazos no deseados e impacte en un grupo poblacional especialmente vulnerable, como lo son las adolescentes y las mujeres jóvenes

Currently there exists increasing preoccupation concerning sexual and reproductive health among teenagers; in spite of the availability of different contraceptive methods, the number of undesired pregnancies is steadily increasing. Among the products presently available for birth control, spermicides are a means that can be totally controlled by the woman and are very reliable compared to other contraceptives in common use. However, they cause irritation in the vaginal epithelium due to their tensoactive effect on cellular membranes which might enhance the risk of acquiring sexually transmissible diseases. In searching for new alternatives, it was observed that a wide variety of plants have spermicidal activity. Hence it is interesting to consider potential contraceptives of vegetable origin, as they may constitute a key tool to prevent undesired pregnancies in general, and in particular in vulnerable groups such as teenagers and young women

Characterization of condom lubricant components using Raman spectroscopy and Raman chemical imaging.

A condom can be described as a protective sheath used as a contraceptive or to protect against sexually transmitted diseases. However, individuals also use condoms during the commission of sexual assaults to prevent identification through deposited biological material. Raman spectroscopy offers a novel approach to identifying the presence of condom lubricant components. Furthermore, Raman chemical imaging expands on conventional Raman spectroscopy to characterize multiple condom lubricant components simultaneously in a manner that effectively demonstrates heterogeneous sample mixtures both spectrally and spatially. Known reference materials, liquid and solid lubricant components of common condom brands were successfully characterized using Raman dispersive spectroscopy and Raman chemical imaging without extensive sample preparation inherent to other analytical methods. The characterization of these materials demonstrates the potential of this technique to become a routine screening method for condom lubricants. This preliminary investigation provides a basis for future studies to determine the feasibility of Raman spectroscopy and Raman chemical imaging for condom lubricant trace detection in case type samples.

Spermicidal & contraceptive properties of Praneem polyherbal pessary.

BACKGROUND & OBJECTIVES: Though a number of barrier methods and potent spermicides are available, most of these have nonoxynol-9 (N-9) as the active ingredient which is observed to cause inflammation and genital ulceration on repeated use. The present study was undertaken to develop a safe spermicide with conjoint microbicidal properties. METHODS: A polyherbal pessary was formulated with purified ingredients from neem (Azadirachta indica) leaves, Sapindus mukerossi (pericarp of fruit) and Mentha citrata oil. Spermicidal action on human sperm was tested by Sander-Cramer slide test in vitro and by post coital tests in vivo. Contraceptive action was tested in rabbits. RESULTS: The combination of the three herbal ingredients resulted in the potentiation of the spermicidal action by 8-folds. The post coital tests confirmed the spermicidal properties of the Praneem polyherbal pessary (PPP) in women with high cervical mucous score around mid estrus. It also prevented in most women the migration of sperm into the cervical mucous. In 15 rabbits studied pregnancy was prevented by the intravaginal administration of PPP, whereas 13 of the 15 animals in the control group became pregnant. INTERPRETATION & CONCLUSION: The Praneem polyherbal pessary has potent spermicidal action on human sperm in vitro and in vivo. When applied in the vagina before mating, it prevented rabbits from becoming pregnant.

Electrospray and MALDI mass spectrometry in the identification of spermicides in criminal investigations.

Electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry have been used to examine evidence in a sexual assault investigation. Because condoms are being used increasingly by sexual assailants and some condom brands include the spermicide nonoxynol-9 (nonylphenoxy polyethoxyethanol) in the lubricant formulation, the recovery, and identification of nonoxynol-9 from evidence items may assist in proving corpus delicti. A method was developed for the recovery of nonoxynol-9 from internal vaginal swabs and for its identification by reverse phase liquid chromatography/electrospray ionization mass spectrometry (LC ESI-MS), nanoelectrospray ionization (nanoESI) mass spectrometry, and high resolution MALDI Fourier transform mass spectrometry (MALDI-FTMS). The method was tested on extracts from precoitus, immediate postcoitus, and four-hours postcoitus vaginal swabs provided by a volunteer whose partner does not normally use condoms, but for this trial used a condom having a water-soluble gel-type lubricant that includes 5% nonoxynol-9 in its formulation. Subsequently, LC ESI-MS was used to identify traces of nonoxynol-9 from the internal vaginal swab of a victim of a sexual assault.

An evaluation of the amount of nonoxynol-9 remaining in the vagina up to 4 h after insertion of a vaginal contraceptive film (VCF) containing 70 mg nonoxynol-9.

The objectives of this study were to determine the amount of nonoxynol-9 (N-9) remaining in the vagina 30 min and 1, 1.5, 2, and 4 h after vaginal insertion of a single sheet of VCF containing 70 mg N-9 and to compare these results to the manufacturer's instructions for use of this product. A new method of vaginal lavage was used to obtain samples for N-9 determination. This was an open-label, noncomparative, pharmacokinetic study in 12 healthy women volunteers not at risk for pregnancy. The study consisted of a screening visit followed by five test visits approximately 1 month apart and a final visit 1 week after all test visits were completed. At each test visit, the investigator inserted a single sheet of VCF in the vagina of the volunteer at midcycle. The volunteer remained in the clinic and underwent vaginal lavage with normal saline after one of five specified time intervals had elapsed. The sequence of the intervals completed by each volunteer was determined by randomization. When undissolved film was found in the vagina, it was removed prior to lavage and assayed for N-9 content separately from that recovered in lavage fluid. It was assumed that the N-9 in undissolved film would not contribute significantly to sperm immobilization. Between 18.5 and 28.5 mg of N-9 were recovered in lavage fluid after intervals of 0.5, 1, 1.5, and 2 h. These levels did not differ statistically (p > 0.05). The amount of N-9 recovered dropped significantly at 4 h to 11.0 mg. If it is assumed that an N-9 concentration of 0.100 mg/mL is required to immobilize sperm in vitro, this study suggests that the amount of N-9 remaining in the vagina in the form of dissolved film up to 4 h after insertion of VCF is sufficient to immobilize sperm. The lavage procedure may not have recovered all N-9 remaining in the vagina. However, intercourse did not take place between insertion and lavage; if it had, the proportion of the film remaining undissolved and the total amount N-9 remaining in the vagina at the time of examination might have been affected.

PIP: The amount of nonoxynol-9 (N-9) remaining in the vagina 0.5, 1.0, 1.5, 2.0, and 4.0 hours after vaginal insertion of a single sheet of a contraceptive film (VCP) containing 70 mg of N-9 was investigated in a pharmacokinetic study involving 12 US women. At each of 5 test visits, approximately 1 month apart, a single sheet of VCF was inserted at midcycle. Vaginal lavage with normal saline was then performed after 1 of the 5 specified time intervals had elapsed. At 30 minutes, an average of 34.4 mg (49% of the total N-9) could be recovered. After intervals of 1.0, 1.5, and 2.0 hours, 18.5-28.5 mg of N-9 was recovered in lavage fluid. The amount of recovered N-9 dropped significantly to 11.0 mg after 4.0 hours. It is assumed that an N-9 concentration of 0.100 mg/mL is required to immobilize sperm. Thus, the amount of N-9 remaining in the vagina up to 4.0 hours after insertion of VCF is sufficient for contraception. The VCF label states that intercourse may take place 15 minutes after film insertion. Although lavage was not performed at this time point, it can be assumed that at least 49% of the original N-9 would be present. Since this study is limited by the fact that intercourse did not take place, future studies should include postcoital measures of the amount of N-9 persisting at various intervals.

Quantitation of vaginally administered nonoxynol-9 in premenopausal women.

A feasibility study was performed in 11 healthy nonpregnant premenopausal women to determine a method for collection and recovery of vaginally administered nonoxynol-9. We also determined if nonoxynol-9 could be quantitated in vaginal lavage fluid obtained 2 h after instillation of a standard precoitol dose of a foam formulation of nonoxynol-9. Samples were analyzed in batch using a validated normal phase high-performance liquid chromatography (HPLC) method. Two hours after instillation of one dose of Delfen Contraceptive Foam (100 mg), the quantity of nonoxynol-9 collected ranged from 10.8 to 67.8 mg (mean: 35.4 mg). This corresponds to a recovery of 11.70%, of the administered dose. Quantitation of vaginally administered nonoxynol-9 is both practical and feasible. These data represent a critical first step in the evaluation of the safety and effectiveness of nonoxynol-9-containing products in the prevention of sexually transmitted diseases.

PIP: In clinical trials, nonoxynol-9 has been shown to protect against the transmission of sexually transmitted pathogens. Conversely, there are concerns that frequent use may lead to vaginal irritation and thus increase the risk of transmission of sexually transmitted diseases (STDs), including human immunodeficiency virus (HIV). Nonoxynol-9 is available as a foam, cream, gel, film, and suppository. These routes of administration differ from each other in their total unit dose, ability to irritate genital tissues, rates of dissolution, and ability to coat the vagina. To determine a method for collection and recovery of vaginally administered nonoxynol-9, and thereby facilitate research on the role of nonoxynol-9 in the prevention of HIV, a feasibility study was performed in 11 healthy, premenopausal US women. Also investigated was whether nonoxynol-9 could be quantitated in vaginal lavage fluid obtained 2 hours after instillation of 100 mg of nonoxynol-9 foam (Delfen). The quantity of nonoxynol-9 collected at this time ranged from 10.8-67.8 mg (mean, 35.4 mg), corresponding to a recovery rate of 11-70% of the original dose. Although further studies are needed to optimize methods for nonoxynol-9 recovery from the vagina, this study suggests it is feasible to quantitate nonoxynol-9 after single-dose vaginal administration.

Determination of the spermicide nonoxynol-9 in vaginal lavage by high-performance liquid chromatography.

A sensitive normal-phase high-performance liquid chromatographic method using a bonded-phase aminosilica column has been developed for the measurement of the spermicide nonoxynol-9 in vaginal lavage fluid. The mean multiple correlation coefficient (r2) for nonoxynol-9 was 0.999 over the calibration range 3.125-50 micrograms/ml for the standards. Quality control samples measured at two different concentration levels gave intra-day precision values (coefficient of variation, C.V.) in the range of 0.61 to 1.63% and the intra-day accuracy values (mean relative error, M.R.E.) in the range of 0.13-0.62%. Inter-day precision and accuracy values from five different calibration standard concentration values ranged from 2.25 to 5.09% C.V. and 4.02 to 7.56% M.R.E. Nonoxynol-9 samples examined for peak area stability at room temperature over a 24-h time period had a M.R.E. of 14.9%. Quality control samples stored at -70 degrees C, and tested after one month by comparison to baseline samples, had a M.R.E. of -10% and 7.53% for the low and high quality control samples, respectively. The method is sensitive and simple, with short runtimes, to enable the processing of numerous samples from a clinical trial.

Extraccion, estudio quimico y determinación de la actividad esparmicida de las saponinas de sapindus saponaria L. (boliche) / Extraction, chemical study and determination of spermicid activity of saponines of sapindus saponaria L. (boliche)

En el presente trabajo se ha realizado la extracción, estudio que por determinación de la actividad espermicida de las saponinas de sapindus saponena L. Boliche. La extracción se llevó a cabo con una macerración previa utilizando una solución hidroalcoholica al 50 porciento luego del desangrado y del despegmentado se extraen las saponinas con n-butonol, obteniendose un rendimiento del 34.08 porciento sobre la muestra deseada. Las saponínas crudas aisladas tienen naturalezas twterpénicos según los procedimientos, estanderes de identificación; el aspecto inflarojo indica que las germinas permanecen al grupo coliano. Los restos azucarados fueron identificados como L (-) ramnosa, L(+) arabinosa y D (+) glucosa utilizando la tecnica cromatográfica en papel. La actividad espermicidad se determino utilizando una modificación de la tecnica de la Baker y del test de Harris, se hallo que las saponinas tiene un primer efecto espermostatico y un posterior efecto espermicida. Tambien se encontró relación en la actividad espermicida y el número de espermatozoides. Los valores de la edad varian según el tiempo de contacto de los espermatozoides con las saponinas.

High-performance liquid chromatographic (HPLC) analysis of oligomeric components of the spermicide nonoxynol-9.

The commercially available Nonoxynol-9 spermicide is a multicomponent mixture of oligomers. When Nonoxynol-9 was separated by normal phase gradient HPLC, 17 components were shown to exist in the commercial mixture. These oligomeric components follow a Poisson distribution around the most abundant oligomer, EO 8 (11.7%). Select oligomers were isolated by preparative HPLC (Rt = 19.6, 34.0, 45.6, 51.2, 61.6, and 79.2 min) and purified by HPLC. These were identified by FAB-MS and NMR to be the oligomers EO 3, EO 6, EO 8, EO 9, EO 11, and EO 16, respectively.

Quantitative analysis of nonoxynol-9 in blood.

At present, there is little data on the level of nonoxynol-9 (N-9) in blood. Using routine methods, the rate of recovery for N-9 in blood has been confirmed to be very low. A method determining N-9 levels in urine has been found but obviously a more efficient method for determining N-9 levels in blood and urine is still needed. Here, a method of determining N-9, a nonionic surfactant spermicide, in blood is presented. The analytical procedure is a simple and convenient method. Using HPLC, UV detector, ODS reversed-phase column, methanol-water mobile phase, NaCl salting-out and two extractions with benzene, a rate of recovery for N-9 of 93 +/- 4% has been obtained.

A simple, sensitive assay for the spermicide nonoxynol-9 in biological fluids by high-performance liquid chromatography.

A new high-performance liquid chromatographic technique has been developed to quantitate nonoxynol-9 in serum, urine, and vaginal fluid. The method is rapid, involves minimal sample preparation, and can be used to analyze a large number of biological fluid samples. The assay elutes a single nonoxynol-9 peak with no interfering components. This was accomplished using a 10-microns pelicular packed amine column, a normal-phase solvent system, and fluorescence detection. Nonoxynol-9 levels as low as 0.23 micrograms/mL in urine can be detected.