Efficient validated method of UPLC-MS/MS to determine curcumin in rat plasma and ovarium

Curcumin, a lipophilic polyphenol derived from the roots of Curcuma longa. Recently, it has been widely investigatedas a therapeutic agent for cancer. Thus, there is a growing interest in measuring curcumin concentrations in theplasma and other target tissues in relevant animal models. We developed and validated a simple, fast, and reliablemethod for quantifying curcumin in biological matrices by Ultra Performance Liquid Chromatography (UPLC)-MassSpectrometry (MS)/MS. The liquid chromatography system is using rapid separation on Acquity UPLC®BEH C18with gradient mobile phase contained formic acid and acetonitrile. Prior to detection, curcumin and internal standard(IS) were ionized using electrospray ionization positive source and the ions were monitored at m/z 369 → 177 and 260→ 183 for curcumin and IS, respectively. The calibration curve was linear (r ≥ 0.99) over the concentration range of1–50 ng/ml and 1–30 ng/ml for rat plasma and for ovary homogenate, respectively. The lower limit of quantificationwas 1 ng/ml. The mean accuracy ranged from 98.9% to 103.2% and 98% to 108.9%, while the coefficient of variation(CV) values of precision in rat plasma were below 11.92% and 10.47% for within and between run, respectively. Inrat ovary homogenate, the mean concentration and CV of within run accuracy and precision were 95.53%–109.78%and 3.34%–9.14%, respectively. The developed method was used to quantify curcumin in rat plasma and ovary afteran oral gavage. In conclusion, the developed and validated method should be useful for quantification of curcuminaccurately and precisely in plasma and target organs from relevant animal models of human diseases.

Survival rate and prognostic factors in advanced cervical cancer patients accompanied by renal impairment.

The objective of this study was to obtain information on the survival rate of advanced cervical cancer patients with renal impairment (ACCRI) and its prognostic factors. In addition, it is hoped that by this method the scoring system for predicting the death of ACCRI patients hopetully could be obtained. Design of the study used was retrospective cohort study. Data collected were retrieved from medical records of ACCRI patients from 1 January 1998 to 31 December 2003 at Dr. Cipto Mangunkusumo National Central General Hospital, Jakarta, with a total sample of 70 cases. The results of the study showed that mean survival of all ACCRI patients was 8.2 months, mean survival at sixth month was 39%, and mean survival at one year was 3.2%. Median survival was 5.3 months. Prognostic factors affecting the survival of ACCRI patients included histopathological type (adenosquamous cell and cell differentiation), cortical thickness of the kidney less than 1 cm, and nephrostomy.

Clinical-pathologic factors, as predictor of lymph nodes metastasis in cervical cancer stage IB and IIA.

The aim of this study was to identify possible predictor factors of lymph node metastases in patients with cervical cancer stage IB and IIA. Study was conducted between May 1996 and December 2001. There were 183 patients of cervical cancer with FIGO Stage IB and IIA who were underwent radical hysterectomy and lymphadenectomy. From those 158 patients could be evaluated, consisting 43 patients with node metastases 115 patients without metastases. Research design was case control study. Case was patients with node metastases and control was those without node metastases. Multivariate analysis was made after bivariate analysis. On bivariate analysis age < 39 years, diameter of lesion > 4 cm, stage IIA > 4 cm, histopathology moderate and poor differentiation, blood and lymphatic vessel invasion were independent variables for node metastases with p value ≤ 0.05. However, on multivariate analysis younger age, parity ≥ 4, diameter of lesion, histopathology adenosquamous, and lymph vascular invasion (+) as independent factors for node metastases with p value ≤ 0.05. Conclusion: Younger age, parity ≥ 4, stage IIA > 4 cm, diameter of lesion, histopathology adenosquamous, and lymph vascular invasion (+) were risk factors for node metastases and can be used as predictors.

Comparison of various procedures in the processing of binucleated-lymphocytes on to glass slide to detect chromosomal breakage.

Binucleated lymphocytes can be screened for micronuclei to assess chromosomal damage. There are various procedures to get slides containing binucleated lymphocytes, that are different in harvesting, fixation, and slide preparation methods. Screening binucleated lymphocytes to find a micronucleus needs at least 800 cells with intact cytoplasm. This study aimed to analyze the various procedures and simplified procedures to know which procedure gave the most abundant binucleated lymphocytes with intact cytoplasm and best staining properties for the purpose of micronucleus scoring. Seven heparinized blood samples were obtained from the Dept. of Obstetrics and gynecology, Faculty of medicine, University of Indonesia, Jakarta. The 7 blood samples were subjected to 17 procedures different in harvesting (with or without washing), slide preparation (smear and spot method, and using a cytocentrifuge), and fixation methods (methanol for 1 minute, methanol brief, methanol/glacial acetic acid 3:1 or 9:1). Our results showed that fixatives containing glacial acetic acid are not suitable for micronucleus test. To generate binucleated lymphocytes with intact cytoplasm as much as possible, the procedure should be conducted without washing steps. Methanol fixation either briefly or 1 minute is preferable, and for the ease of screening cytocentrifuge preparation, followed by spot method is preferable.