浓缩、精制及干燥对复方丹参提取液中水溶性成分的影响

目的：筛选出最佳的以丹参为君药的复方中药水提取液的浓缩、精制及干燥工艺及其条件，以使丹参水溶性酚酸尽可能少地被破坏，且其工艺简单、成本低，适宜于大生产。方法：以丹参中丹酚酸为筛选指标，对其精制、浓缩及干燥过程中的工艺条件进行筛选。结果：以90℃减压沈缩，2500r/min、30min离心精制，进口温度为140℃喷雾干燥为最佳的浓缩、精制及干燥条件。结论：该工艺合理、可行，水溶性酚酸得以最大限度地被保留。     

A 1-year time course study of human RNA degradation in body fluids under dry and humid environmental conditions

Blood, saliva and semen are some of the forensically most relevant biological stains found at crime scenes. mRNA profiling is a reliable approach for the identification of the origin of an evidentiary trace. A stable set of markers and the knowledge about the effects of RNA degradation under different environmental conditions is necessary for the determination of an unknown biological stain. The aim of this work was to compare RNA degradation for human blood, semen and saliva at three different concentrations during a 1-year time period and exposed to dry and humid conditions. Also, this study addressed the question whether there are relevant differences in the efficiency of two RNA extraction methods.     

Evaluation of amylase testing as a tool for saliva screening of crime scene trace swabs

Amylase testing has been used as a presumptive test for crime scene saliva for over three decades, mainly to locate saliva stains on surfaces. We have developed a saliva screening application for crime scene trace swabs, utilising an amylase sensitive paper (Phadebas^® Forensic Press test). Positive results were obtained for all tested dried saliva stains (0.5–32 μL) with high or intermediate amylase activity (840 and 290 kU/L). Results were typically obtained within 5 min, and all samples that produced DNA profiles were positive. However, salivary amylase activities, as well as DNA concentrations, vary significantly between individuals. We show that there is no correlation between amylase activity and amount of DNA in fresh saliva. Even so, a positive amylase result indicates presence of saliva, and thereby presence of DNA. Amylase testing may be useful for screening in investigations where the number of DNA analyses is limited due to cost, e.g., in volume crime.     

Oral fluid collection: The neglected variable in oral fluid testing

The potential to use oral fluid as a drug-testing specimen has been the subject of considerable scientific interest. The ease with which specimens can be collected and the potential for oral fluid (OF) drug concentrations to reflect blood–drug concentrations make it a potentially valuable specimen in clinical as well as forensic settings. However, the possible effects of the OF collection process on drug detection and quantification has often been over looked. Several studies have documented that drug-contamination of the oral cavity may skew oral fluid/blood drug ratios and confound interpretation when drugs are smoked, insufflated or ingested orally. OF pH is predicted to have an effect on the concentration of drugs in OF. However, in a controlled clinical study, the effect of pH was less than that of collection technique. Mean codeine OF concentrations in specimens collected a non-stimulating control method were 3.6 times higher than those in OF collected after acidic stimulation. Mean codeine concentrations were 50% lower than control using mechanical stimulation and 77% of control using commercial collection devices.Several factors should be considered if a commercial OF collection device is used. In vitro collection experiments demonstrated that the mean collection volume varied between devices from 0.82 to 1.86 mL. The percentage of the collected volume that could be recovered from the device varied from 18% to 83%. In vitro experiments demonstrated considerable variation in the recovery of amphetamines (16–59%), opiates (33–50%), cocaine and benzoylecgonine (61–97%), carboxy-THC (0–53%) and PCP (9–56%). Less variation in collection volume, volume recovered and drug recovery was observed intra-device. The THC stability was evaluated in a common commercial collection protocol. Samples in the collection buffer were relatively stable for 6 weeks when stored frozen. However, stability was marginal under refrigerated conditions and poor at room temperature. Very little has been published on the efficacy of using IgG concentration, or any other endogenous marker, as a measure of OF specimen validity. Preliminary rinsing experiments with moderate (50 mL and 2 × 50 mL) volumes of water did not reduce the OF IgG concentration below proposed specimen validity criteria. In summary, obvious and more subtle variables in the OF collection may have pronounced effects on OF–drug concentrations. This has rarely been acknowledged in the literature, but should to be considered in OF drug testing, interpretation of OF–drug results and future research studies.     

唾液中Lewis血型物质的定量研究

应用时间决定性荧光免疫测定法（TR－FIA法），对129例健康成人唾液中Lweis及H1血型物质进行定量检测。Le阳性个体均不同程度地检出了Lea和Leb物质。Lea物质含量：Le（a＋b-）型＞Le（a－b＋）型；Leb物质含量：Le（a－b＋）型＞Le（a＋b－）型。Le（a＋b－）型的Lea物质＞Leb，Le（a－b＋）型的Leb物质＞Lea物质。Le阴性的部分个体未能检出Lewis物质，其余个体也仅检出微量。根据Lea和Leb物质在唾液中的相对含量，可以推测红细胞Lewis型，并提示Leb物质可能由Lea物质转化而形成。     

The killer outfit and timing: Impact of the fabric and time in body fluid identification and DNA profiling

The present work aimed to study the detection, through lateral flow immunochromatographic (LFI) tests, of saliva samples over time in three different types of fabrics, as well as, the possibility of DNA isolation and characterization from the sample tubes and the cassettes. Fifty microliters of saliva (three samples/time) were deposited in denim, cotton, and polyester. Saliva was identified by SERATEC® Amylase Test and the Crime Scene version SALIVA CS, being able to detect it up to six months of deposition, although with different band intensities. Polyester showed stronger bands than cotton, probably due to its synthetic nature, and denim, as an inked fabric, showed less band intensities. Statistical analyses confirmed significant differences among fabrics, but not over time in the same type of fabric. Total DNA from the sample tubes was successfully recovered, in contrast, from the cassettes, only polyester retrieved amplifiable DNA. These findings indicated that it is possible to recover and identify saliva up to six months after deposition, also obtaining DNA. Future research will be able to expand these results, analyzing the stability of other body fluids, and the sensitivity of lateral flow immunochromatographic tests to detect them.     

Oral fluid testing for driving under the influence of drugs: history, recent progress and remaining challenges

In recent years the demand for drug testing in oral fluid in cases of driving under the influence has been increasing. The main advantages of saliva/oral fluid are the possibility for non-medical personnel to collect it without embarrassment and a better correlation between presence of drugs in oral fluid and impairment. Several surveys have been performed since the 1980s using saliva, and researchers encountered problems related to insufficient sample volume and insufficient sensitivity of the analytical methods. Steady progress has been shown in sample collection, knowledge of toxicokinetics in oral fluid, reliability of on-site and laboratory-based immunoassays and confirmation methods. In a few countries, legislation was passed that allows the use of saliva as a matrix for screening or confirmation.Despite this progress, some more work needs to be done, principally in the areas of the sensitivity and reliability of on-site screening devices, particularly for cannabis and benzodiazepines, knowledge about passive contamination and more generalised proficiency testing before oral fluid testing for DUID will have the reliability needed to be used forensically.     

Comparative performance of the Phadebas® Forensic Press Test at room temperature and 37 °C for the detection of saliva stains on fabric exhibits

The Phadebas® Forensic Press Test (PFPT) is an enzyme-based colorimetric test used to visualise and locate latent saliva stains on forensic exhibits. The test relies upon the presence of the enzyme α-amylase which is present in high levels in saliva. Even though the optimal in vitro temperature for α-amylase activity is 37 °C, the PFPT manufacturer’s protocol specifies that the PFPT should be carried out at room temperature (RT). In this study, we compared the performance of the PFPT at RT and 37 °C using combinations of four fabric types (cotton, polyester, acrylic and a cotton/polyester blend), three saliva dilutions (neat, 1:10 and 1:100) and stains aged for four time periods (1 day, 1 week, 1 month and 3 months). The intensity of the PFPT colour reactions at RT and 37 °C were not statistically different across all fabric types, saliva concentrations and stain ages, indicating that maximum sensitivity and performance of the PFPT can be achieved at RT.     

人唾液中苯丙胺类药物检测方法进展

目前人唾液中苯丙胺类药物的检测研究较少,鉴于唾液检材具备很多的实用价值和优势,本文综述了国内外对唾液检材中苯丙胺类药物的检测研究进展,重点包括免疫分析筛选方法、液-液萃取法、固相微萃取法等萃取方法以及气相色谱/质谱联用法、液相色谱/质谱联用法等确证方法的发展情况。